I hope to publish and present a paper at the "Automated Mineralogy" conference this year (Sept 1-2, Brisbane, Oz). Part of it will be a section on error analysis, and I would like to include examples of similar determinations as accomplished by other individuals. That is, there is no independent check on this type of measurement, and even I judged the quality of my measurements visually. It is a very simple task using tools as provided by all image analysis softwares, which involves segmentation of a specific mineral in the presence of other minerals. I have posted an example image here: http://www.micro-investigations.com/rrr.htm (It is an JPEG example only, and I haven't yet determined the appropriate image)
This message is only intended to gauge the interest is such a project, but I do imagine that some of you will recognize immediately the problems associated with this image (and tens of others like it acquired over a short period of time with an SEM), and who may also like to contribute beyond measuring a single image. For example, my script has measured several thousand of these images, and for this paper I would surely be interested in input beyond sampling as many as possible individuals' visual acuity with a single image. I still need to consult my co-author, but significant contributions could mean adding co-authors ... minimally being acknowledged.
Thank you in advance for your interest (or criticisms) ... Genuinely, Michael Shaffer :o)
SEM/MLA Research Coordinator http://www.mun.ca/creait/maf/ Inco Innovation Centre Memorial University St. John's, Newfoundland Canada
==============================Original Headers============================== 7, 18 -- From michael-at-shaffer.net Sat Mar 31 14:13:04 2007 7, 18 -- Received: from n034.sc1.he.tucows.com (smtpout1474.sc1.he.tucows.com [64.97.157.174]) 7, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l2VJD4E7004158 7, 18 -- for {Microscopy-at-microscopy.com} ; Sat, 31 Mar 2007 14:13:04 -0500 7, 18 -- Received: from rarewolf (205.251.83.78) by n034.sc1.he.tucows.com (7.2.069.1) (authenticated as michael-at-shaffer.net) 7, 18 -- id 45B9475500D3B8B4 for Microscopy-at-microscopy.com; Sat, 31 Mar 2007 19:12:59 +0000 7, 18 -- From: "michael shaffer" {michael-at-shaffer.net} 7, 18 -- To: "MSA Microscopy list" {Microscopy-at-microscopy.com} 7, 18 -- Subject: Image Analysis: round robin measurement 7, 18 -- Date: Sat, 31 Mar 2007 16:40:51 -0230 7, 18 -- Message-ID: {006d01c773c8$4d3104a0$4701a8c0-at-rarewolf} 7, 18 -- MIME-Version: 1.0 7, 18 -- Content-Type: text/plain; 7, 18 -- charset="us-ascii" 7, 18 -- Content-Transfer-Encoding: 7bit 7, 18 -- X-Mailer: Microsoft Office Outlook 11 7, 18 -- Thread-Index: AcdzyExcHUYfF8CGT++Z2CZOtX+sqg== 7, 18 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.3028 ==============================End of - Headers==============================
Regarding my previous post to the list, I should have asked that you contact me directly, except and unless you feel your reply is of general interest ...
Genuinely, Michael Shaffer :o)
SEM/MLA Research Coordinator http://www.mun.ca/creait/maf/ Inco Innovation Centre Memorial University St. John's, Newfoundland Canada
==============================Original Headers============================== 6, 18 -- From michael-at-shaffer.net Sat Mar 31 14:15:53 2007 6, 18 -- Received: from n034.sc1.he.tucows.com (smtpout1474.sc1.he.tucows.com [64.97.157.174]) 6, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l2VJFqBc007169 6, 18 -- for {Microscopy-at-microscopy.com} ; Sat, 31 Mar 2007 14:15:53 -0500 6, 18 -- Received: from rarewolf (205.251.83.78) by n034.sc1.he.tucows.com (7.2.069.1) (authenticated as michael-at-shaffer.net) 6, 18 -- id 45B9475500D3B97F for Microscopy-at-microscopy.com; Sat, 31 Mar 2007 19:15:52 +0000 6, 18 -- From: "michael shaffer" {michael-at-shaffer.net} 6, 18 -- To: "MSA Microscopy list" {Microscopy-at-microscopy.com} 6, 18 -- Subject: Image Analysis: round robin measurement (oops!) 6, 18 -- Date: Sat, 31 Mar 2007 16:43:45 -0230 6, 18 -- Message-ID: {006e01c773c8$b48336a0$4701a8c0-at-rarewolf} 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 -- Thread-Index: AcdzyExcHUYfF8CGT++Z2CZOtX+sqg== 6, 18 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.3028 ==============================End of - Headers==============================
Your suggestion brings up an interesting point. That is, I haven't yet created my "instructions" message that would outline what I want and in what form, but it should surely invite other methods other than "grayscale segmentation" as improperly suggested as the only method applicable.
Genuinely, Michael Shaffer :o)
SEM/MLA Research Coordinator http://www.mun.ca/creait/maf/ Inco Innovation Centre Memorial University St. John's, Newfoundland
} -----Original Message----- } From: Webster, Paul [mailto:PWebster-at-hei.org] } Sent: March 31, 2007 6:29 PM } To: michael-at-shaffer.net } Subject: RE: [Microscopy] Image Analysis: round robin measurement } } Hi Micheal, } } It looks like you are seeking a way to get what biologists } know as a volume density analysis. We do it very simply by } placing cross-lattice overlays onto the iamges and then } counting the number of points over the full structure and } then counting points only over the strucuture of interest. } } Weibel first introduced the method and it has been verified } by many people applying it to many applications. Hans } Gundersen and Terry Mayhew have written a few reviews of the } method. StereoInvestigator (MicroBrightField, in the US) and } the CAST system (Olympus) have semi-automated it. } } It is a rapid, accurate way of estimating volume densities } and if performed correctly gives a estimate with about a 5% } error. At least is does seem to be that way for our } biological samples. } } Regards, } } Paul Webster. } } } } } -----Original Message----- } From: michael-at-shaffer.net [mailto:michael-at-shaffer.net] } Sent: Sat 3/31/2007 12:19 PM } To: Webster, Paul } Subject: [Microscopy] Image Analysis: round robin measurement } } } } } -------------------------------------------------------------- } -------------- } 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 :o) } } I hope to publish and present a paper at the "Automated Mineralogy" } conference this year (Sept 1-2, Brisbane, Oz). Part of it } will be a section on error analysis, and I would like to } include examples of similar determinations as accomplished by } other individuals. That is, there is no independent check on } this type of measurement, and even I judged the quality of my } measurements visually. It is a very simple task using tools } as provided by all image analysis softwares, which involves } segmentation of a specific mineral in the presence of other } minerals. I have posted an example image here: } http://www.micro-investigations.com/rrr.htm } (It is an JPEG example only, and I haven't yet determined the } appropriate } image) } } This message is only intended to gauge the interest is such a } project, but I do imagine that some of you will recognize } immediately the problems associated with this image (and tens } of others like it acquired over a short period of time with } an SEM), and who may also like to contribute beyond measuring } a single image. For example, my script has measured several } thousand of these images, and for this paper I would surely } be interested in input beyond sampling as many as possible } individuals' visual acuity with a single image. I still need } to consult my co-author, but significant contributions could } mean adding co-authors ... minimally being acknowledged. } } Thank you in advance for your interest (or criticisms) ... } Genuinely, Michael Shaffer :o) } } SEM/MLA Research Coordinator } http://www.mun.ca/creait/maf/ } Inco Innovation Centre } Memorial University } St. John's, Newfoundland } Canada } } } } ==============================Original } Headers============================== } 7, 18 -- From michael-at-shaffer.net Sat Mar 31 14:13:04 2007 7, } 18 -- Received: from n034.sc1.he.tucows.com } (smtpout1474.sc1.he.tucows.com [64.97.157.174]) } 7, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) } with ESMTP id l2VJD4E7004158 } 7, 18 -- for {Microscopy-at-microscopy.com} ; Sat, 31 Mar } 2007 14:13:04 -0500 } 7, 18 -- Received: from rarewolf (205.251.83.78) by } n034.sc1.he.tucows.com (7.2.069.1) (authenticated as } michael-at-shaffer.net) } 7, 18 -- id 45B9475500D3B8B4 for } Microscopy-at-microscopy.com; Sat, 31 Mar 2007 19:12:59 +0000 } 7, 18 -- From: "michael shaffer" {michael-at-shaffer.net} 7, 18 } -- To: "MSA Microscopy list" {Microscopy-at-microscopy.com} 7, } 18 -- Subject: Image Analysis: round robin measurement 7, 18 } -- Date: Sat, 31 Mar 2007 16:40:51 -0230 7, 18 -- Message-ID: } {006d01c773c8$4d3104a0$4701a8c0-at-rarewolf} } 7, 18 -- MIME-Version: 1.0 } 7, 18 -- Content-Type: text/plain; } 7, 18 -- charset="us-ascii" } 7, 18 -- Content-Transfer-Encoding: 7bit 7, 18 -- X-Mailer: } Microsoft Office Outlook 11 7, 18 -- Thread-Index: } AcdzyExcHUYfF8CGT++Z2CZOtX+sqg== 7, 18 -- X-MimeOLE: Produced } By Microsoft MimeOLE V6.00.2900.3028 } ==============================End of - } Headers============================== } } }
==============================Original Headers============================== 8, 21 -- From michael-at-shaffer.net Sun Apr 1 05:58:12 2007 8, 21 -- Received: from n007.sc1.he.tucows.com (smtpout1443.sc1.he.tucows.com [64.97.157.143]) 8, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l31AwCEt022218 8, 21 -- for {Microscopy-at-microscopy.com} ; Sun, 1 Apr 2007 05:58:12 -0500 8, 21 -- Received: from rarewolf (205.251.83.78) by n007.sc1.he.tucows.com (7.2.069.1) (authenticated as michael-at-shaffer.net) 8, 21 -- id 45B8E5A000D7B24B; Sun, 1 Apr 2007 10:58:11 +0000 8, 21 -- From: "michael shaffer" {michael-at-shaffer.net} 8, 21 -- To: "'Webster, Paul'" {PWebster-at-hei.org} 8, 21 -- Cc: "MSA Microscopy list" {Microscopy-at-microscopy.com} 8, 21 -- References: {200703311919.l2VJJOd0017547-at-ns.microscopy.com} {87449E4A2B01DA47B29424CE5D6E0F8304178D7E-at-hi0sml1.hei.org} 8, 21 -- Subject: RE: [Microscopy] Image Analysis: round robin measurement 8, 21 -- Date: Sun, 1 Apr 2007 08:26:03 -0230 8, 21 -- Message-ID: {008301c7744c$58323390$4701a8c0-at-rarewolf} 8, 21 -- MIME-Version: 1.0 8, 21 -- Content-Type: text/plain; 8, 21 -- charset="us-ascii" 8, 21 -- Content-Transfer-Encoding: 7bit 8, 21 -- X-Mailer: Microsoft Office Outlook 11 8, 21 -- In-Reply-To: {87449E4A2B01DA47B29424CE5D6E0F8304178D7E-at-hi0sml1.hei.org} 8, 21 -- Thread-Index: AcdzyX8cRddFIXwhQren9CNjXF0otgADSu5GAB0+njA= 8, 21 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.3028 ==============================End of - Headers==============================
We have a Varian 880 high-vac ionization gauge that works fine, as far as I know. At least it did when we pulled it off the TEM it was on.
We also have an Arkay Gas Burst timer in basically mint condition. It was a spare for one of our nitrogen-burst processors, so I can't say for sure that it works, but it looks like it's right out of the box.
You payeth for shipping, they belongeth to you.
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 2 13:26:20 2007 7, 23 -- Received: from um-tsmtpout1.um.umsystem.edu (um-tsmtpout1.um.umsystem.edu [209.106.228.23]) 7, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l32IQKsb004731 7, 23 -- for {microscopy-at-microscopy.com} ; Mon, 2 Apr 2007 13:26:20 -0500 7, 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); 7, 23 -- Mon, 2 Apr 2007 13:26:19 -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: Free stuff 7, 23 -- Date: Mon, 2 Apr 2007 13:26:19 -0500 7, 23 -- Message-ID: {91108EF9255B394CBF8B7E3789814A4101C0B89F-at-UM-XMAIL08.um.umsystem.edu} 7, 23 -- X-MS-Has-Attach: 7, 23 -- X-MS-TNEF-Correlator: 7, 23 -- Thread-Topic: Free stuff 7, 23 -- Thread-Index: Acd1VGhsnFdoSu9tR3Gb5vuMqMMW3Q== 7, 23 -- From: "Tindall, Randy D." {TindallR-at-missouri.edu} 7, 23 -- To: {microscopy-at-microscopy.com} 7, 23 -- X-OriginalArrivalTime: 02 Apr 2007 18:26:19.0782 (UTC) FILETIME=[68B62A60:01C77554] 7, 23 -- Content-Transfer-Encoding: 8bit 7, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l32IQKsb004731 ==============================End of - Headers==============================
Sorry, all. In my urge to clean up the lab, I typed before I thought.
It happens that these items in my previous post will need to go through our surplus property system, rather than through an informal give-away.
My apologies for any disappointments.
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============================== 6, 23 -- From TindallR-at-missouri.edu Mon Apr 2 13:35:11 2007 6, 23 -- Received: from um-nsmtpout1.um.umsystem.edu (um-nsmtpout1.um.umsystem.edu [209.106.228.53]) 6, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l32IZBKW016253 6, 23 -- for {microscopy-at-microscopy.com} ; Mon, 2 Apr 2007 13:35:11 -0500 6, 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); 6, 23 -- Mon, 2 Apr 2007 13:35:11 -0500 6, 23 -- x-mimeole: Produced By Microsoft Exchange V6.5 6, 23 -- Content-class: urn:content-classes:message 6, 23 -- MIME-Version: 1.0 6, 23 -- Content-Type: text/plain; 6, 23 -- charset="us-ascii" 6, 23 -- Subject: Free stuff retraction 6, 23 -- Date: Mon, 2 Apr 2007 13:35:10 -0500 6, 23 -- Message-ID: {91108EF9255B394CBF8B7E3789814A4101C0B8A1-at-UM-XMAIL08.um.umsystem.edu} 6, 23 -- X-MS-Has-Attach: 6, 23 -- X-MS-TNEF-Correlator: 6, 23 -- Thread-Topic: Free stuff retraction 6, 23 -- Thread-Index: Acd1VaU5PYZ7gSqgSh6Zb6+Qw1G7gQ== 6, 23 -- From: "Tindall, Randy D." {TindallR-at-missouri.edu} 6, 23 -- To: {microscopy-at-microscopy.com} 6, 23 -- X-OriginalArrivalTime: 02 Apr 2007 18:35:11.0351 (UTC) FILETIME=[A58D2870:01C77555] 6, 23 -- Content-Transfer-Encoding: 8bit 6, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l32IZBKW016253 ==============================End of - Headers==============================
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (m_bouchaour-at-yahoo.fr) from http://www.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Tuesday, April 3, 2007 at 04:01:39 ---------------------------------------------------------------------------
Email: m_bouchaour-at-yahoo.fr Name: mama bouchaour
Organization: university of metz
Education: Graduate College
Location: metz, france
Question: while i scanned a surface of GaN/LiNbO3 in SEM, i saw, under the layer of GaN a white and brillant zone anb below the bulk of liNbO3. Why this white region appear. GaN is a semiconductor and LiNbO3 is very resistive.
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 dieter.bosshardt-at-zmk.unibe.ch as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Organization: School of Dental Medicine, University of Berne
Title-Subject: [Filtered] TRAP staining in thick MMA ground sections
Question: We would like to do tartrate-resistant acid phosphatase (TRAP) staining to visualize osteoclasts in methylmethacrylate (MMA)-embedded tissues. Most people do TRAP staining in paraffin sections or in about 5 micron thick MMA sections. In the past, we successfully did TRAP staining in about 100 micron thick MMA (ground) sections. However, at the moment we are unable to repeat the TRAP staining in thick MMA ground sections. Is there anyone out there who has a protocol for TRAP staining in thick MMA ground sections? We fixed the tissues in 4% formalin. Thanks a lot in advance.
Well, we received the SEM today. We got it off of the truck and into the cafeteria, which we are using as a staging area until we can get it into its permanent home in my lab upstairs. That's right, upstairs.
Despite their best efforts to the contrary, the shipping company did not damage it too much. The main table holding the chamber had shifted off of the springs, but was still fairly stable when we unpacked it. The main transformer in the bottom of the control console had leaked all over, so we are dismantling the console to get all the oil out. Most of the screws had fallen out of the column itself, so we just marked the location and order of the sections and took them off as we moved it into the cafeteria. I found a good number of the screws stuck to the side of the ion pump magnet which had been set on the bottom of the main scope unit.
The vacuum system is a disaster, though. Whoever disconnected this SEM from service did a real hack job. The water hoses are cut, and the vacuum system was almost entirely dismantled. All of the compressed air/nitrogen lines were disconnected. Anybody have a good schematic as to which valves get connected where?
So, here is the order of the project priorities:
1: Get everything upstairs. Most will be in pieces when it gets up there, but we have documented it to no end (About 180 photos of the unit all taken down)
2: Re-assemble the column and microscope console. This will include re-working the entire vacuum system. There is oil in the diffusion pumps, but I don't know how much. I'll check and see if it pumps before trying to add more. There is a measurable amount, so I'm not too worried. It looks clean and good. I need to get all the water and air lines connected again. Again- if anybody has a schematic of this system I would greatly appreciate it.
3: Open and clean out the main transformer tank. Replace the oil, since most of it is in the bottom of the truck and control console.
4: Re-connect everything in the control console. Luckily most of the cables are labeled well.
5: Figure out the six wires that were cut. There are six wires that were cut between the center triangular console and wherever they went. Luckily, though, I can see that they have different numbers of wires within the cables, so I should be able to get a decent idea where they go when I find the other end. Then, the six conductor wire gets spliced back to the six conductor, etc...
6: Obtain a vacuum pump. There was no pump box for this system.
7: Test the vacuum seals. Since we are having to re-seal most of the vacuum connections, I will run several dry runs with the vacuum system. I don't see an indicator for the vacuum level anywhere on the console. Is there a test point I can read the value off of?
8: Order new filaments. None were included. I think I'll add a set of apertures to that as well.
9: Try to power it up and see what happens at low accelerating voltages.
10: Explore the microscopic world! (We hope.)
It looks as though this was at one point hooked up to a digital acquisition system, as there are soldered on BNC connectors labeled for horizontal, vertical, and beam control.
One final question: In the control console, there are two boxes at the bottom most level. The one on the left (As you are looking at the rear) is the main HV transformer tank. The one on the right looks like it has some plug-in modules in it, and there is a hose running from each module to the next module. Is this for cooling water or oil? There were no connections or hoses that lead to the system or away from it.
This Question was submitted to Ask-A-Microscopist by (marilyn.lemieux-at-genzyme.com) from http://microscopy.org/Ask-A-Microscopist/Ask-A-Microscopist.html on Tuesday, April 3, 2007 at 12:33:59 Remember to consider the Grade/Age of the student when considering the Question --------------------------------------------------------------------------- Please reply to both marilyn.lemieux-at-genzyme.com as well as to the Microscopy Listserver ---------------------------------------------------------------------------
Email: marilyn.lemieux-at-genzyme.com Name: Marilyn LeMieux
Organization: Genzyme Genetics
Education: Graduate College
Location: Orange, CA, USA
Title: Koehler illumination
Question: Some books say that this must be performed on both low and high power as you focus on an object to be imaged (digital image). Others say that you only need to perform the koehler steps on High power(images are taken only on high power). Do those who say to do it on both low, then high power, is that to 'get you in the ballpark' prior to going to high, or is this step unnecessary?
I am trying to simulate electron beam heating in the SEM. I am sure this is not a new topic and perhaps lots of people had done some work on it. I am totally new to this area so like to check if anyone has good journals to recommend?
In my simulation, I input a figure for the probe current density (got from some journals), I inevitably gets melting... I am still trying to verify this.
Can anyone point out to me a typical figure for probe size, current and perhaps even current distribution equation for a TEM probe?
Thank you very much
Regards TT
==============================Original Headers============================== 3, 23 -- From tttan-at-SIMTech.a-star.edu.sg Tue Apr 3 20:18:45 2007 3, 23 -- Received: from smtp1.simtech.a-star.edu.sg (smtp1.SIMTech.a-star.edu.sg [203.126.240.71]) 3, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l341Iisr007682 3, 23 -- for {Microscopy-at-microscopy.com} ; Tue, 3 Apr 2007 20:18:45 -0500 3, 23 -- Received: from marlin.SIMTech.a-star.edu.sg ([203.125.167.16]) 3, 23 -- by smtp1.simtech.a-star.edu.sg (8.13.8/8.13.8) with ESMTP id l341Iaei021427 3, 23 -- for {Microscopy-at-microscopy.com} ; Wed, 4 Apr 2007 09:18:36 +0800 3, 23 -- X-MimeOLE: Produced By Microsoft Exchange V6.0.6603.0 3, 23 -- content-class: urn:content-classes:message 3, 23 -- MIME-Version: 1.0 3, 23 -- Content-Type: text/plain; 3, 23 -- charset="iso-8859-1" 3, 23 -- Subject: RE: Electron beam simulation 3, 23 -- Date: Wed, 4 Apr 2007 09:18:35 +0800 3, 23 -- Message-ID: {04E75CD46E6CD64A804C465D4CA0E48FA395DB-at-marlin.simtech.a-star.edu.sg} 3, 23 -- X-MS-Has-Attach: 3, 23 -- X-MS-TNEF-Correlator: 3, 23 -- Thread-Topic: Electron beam simulation 3, 23 -- Thread-Index: Acd0xxCViVhBKNoQRY+l2Ux1UmpcowBkA1RA 3, 23 -- From: "Tan Thiam Teck, Dr" {tttan-at-SIMTech.a-star.edu.sg} 3, 23 -- To: {Microscopy-at-microscopy.com} 3, 23 -- Content-Transfer-Encoding: 8bit 3, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l341Iisr007682 ==============================End of - Headers==============================
I don't understand your expression, but performing a Koehler takes less than 1 min, so why bother about NOT doing it at lower mag if this can help at higher mag? If you think this does not help, why do you care? The purpose it to focus the beam on your object, in the condition you take the picture. That said, if you take all your pictures are the same (high) magnification, you probably don't need to perform a Koehler each and every time.
Stephane
--- marilyn.lemieux-at-genzyme.com wrote:
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To quote: "The Koehler technique is recommended by all manufacturers of modern laboratory microscopes because it can produce specimen illumination that is uniformly bright and free from glare, thus allowing the user to realize the microscope's full potential."
Note that you should check Koehler illumination this every-time you change objective on a microscope, and setting Koehler illumination is crucial if you are using Phase Contrast (or DIC) optical contrast enhancement. So even low power phase objectives require Koehler adjustment for good images via transmission illumination. It is also required if you are capturing transmission images via a camera (or they won't look that good at all).
For heavily stained sections at low magnifications you can get by without bothering, but as Stephane points out it takes very little time to setup and it is poor science not to check it every time you use the microscope (particularly as you will have spent many hours preparing the specimen). Previous users may have setup the optics incorrectly for various reasons.
Koehler illumination is irrelevant with epi-fluorescent imaging as with this the light is backscattered into the objective, although often you will also want a standard phase contrast or DIC transmission image as well. Koehler illumination is essential for transmission images of unstained specimens with limited contrast (where phase contrast or DIC optics is often also required to enhance the specimens contrast by optical interference within structures inside the specimen).
Poorly adjusted optics lead to very uneven illumination and the appearance of dark shadows in the image. It will very badly affect contrast enhancement optics (you won't get much enhancement). These problems are naturally best avoided, particularly as setting the optics correctly is so easy. All microscope manuals will tell you how to set up Kohler illumination with the microscope (plus other important things like aligning illumination bulbs and phase contrast rings). Expensive modern motorised microscopes can do much of this automatically these days.
Regards
Keith
--------------------------- Dr Keith J Morris Imaging Facilities Manager Cell Biology Institute of Ophthalmology UCL, London EC1V 9EL
-----Original Message----- X-from: marilyn.lemieux-at-genzyme.com [mailto:marilyn.lemieux-at-genzyme.com] Sent: 04 April 2007 02:18 To: keith.morris-at-ucl.ac.uk
This Question was submitted to Ask-A-Microscopist by (marilyn.lemieux-at-genzyme.com) from http://microscopy.org/Ask-A-Microscopist/Ask-A-Microscopist.html on Tuesday, April 3, 2007 at 12:33:59 Remember to consider the Grade/Age of the student when considering the Question --------------------------------------------------------------------------- Please reply to both marilyn.lemieux-at-genzyme.com as well as to the Microscopy Listserver ---------------------------------------------------------------------------
Email: marilyn.lemieux-at-genzyme.com Name: Marilyn LeMieux
Organization: Genzyme Genetics
Education: Graduate College
Location: Orange, CA, USA
Title: Koehler illumination
Question: Some books say that this must be performed on both low and high power as you focus on an object to be imaged (digital image). Others say that you only need to perform the koehler steps on High power(images are taken only on high power). Do those who say to do it on both low, then high power, is that to 'get you in the ballpark' prior to going to high, or is this step unnecessary?
If you drive a car with a manual gear shift when you want to go, you push in the clutch, put it into first gear and let out the clutch; then you push in the clutch put it into second gear and let out the clutch, then you push in the clutch and put it into third gear and let out the clutch.
When you are driving a manual microscope, you click in the low power objective, change the diaphragm and adjust the condenser; when you want middle magnification (second gear) you change the diaphragm and adjust the condenser; and when you want high power, etc.
I can continue to play with this analogy. I do not know why people can accept moving the focus adjustment on a microscope but not the condenser adjustment when magnification is changed. Perhaps these poor souls did not do well in optics when they took college physics.
Bob Blystone
On Apr 4, 2007, at 2:21 AM, nizets2-at-yahoo.com wrote:
} } Hi! } } I don't understand your expression, but performing a } Koehler takes less than 1 min, so why bother about NOT } doing it at lower mag if this can help at higher mag? } If you think this does not help, why do you care? The } purpose it to focus the beam on your object, in the } condition you take the picture. } That said, if you take all your pictures are the same } (high) magnification, you probably don't need to } perform a Koehler each and every time. } } Stephane } } } } } Title: Koehler illumination } } } } Question: Some books say that this must be performed } } on both low and high power as you focus on an object } } to be imaged (digital image). Others say that you } } only need to perform the koehler steps on High } } power(images are taken only on high power). } } Do those who say to do it on both low, then high } } power, is that to 'get you in the ballpark' prior to } } going to high, or is this step unnecessary?
Dear Marilyn, It is always good to "Kohler" every time you change lenses, especially if you are going to take pictures (digital or otherwise). "Kohlering" aligns the illumination system with the rest of the microscope's optical axis, ensuring even illumination without odd shadings or shadows. Once you get the hang of it, it only takes a few seconds to do it, so it is certainly worth the effort. Lee -- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy & Histology Core Facility Manager, Optical Microscopy Core Facility Weill-Cornell Medical College
Dear Marilyn, It is always good to "Kohler" every time you change lenses, especially if you are going to take pictures (digital or otherwise). "Kohlering" aligns the illumination system with the rest of the microscope's optical axis, ensuring even illumination without odd shadings or shadows. Once you get the hang of it, it only takes a few seconds to do it, so it is certainly worth the effort. 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 can't stress strongly enough the importance of establishing Koehler illumination for all techniques ... and I agree strongly with several of the responses which stress how easy and quick the process is, once you have done it a few times.
Koehler illumination establishes the "baseline" for all other imaging. Setting aside alignment of the lamp filament (which typically only needs to be done when the lamp is changed), it involves the simple setting of focus and apertures for three key lens sets: objective, condenser, and eyepieces. On most microscopes, each of these lenses has adjustment for focus. Also, it is important to understand the appropriate setting for the field iris (which controls scatter and glare) and aperture iris (which controls coherence and has a major impact on edge fidelity as well as resolution).
Unfortunately, today's schedule doesn't permit a long discussion, but for those of you who are interested in a brief anatomny and physiology less regarding each of the three key lenses and their apertures... Plus a short recipe for establishing Koehler, send me an email with KOEHLER, PLEASE in the subject and I'll try to send you a PDF early next week, when I am back in the office. Also, there is a detailed section in Optimizing Light Microscopy (call Ken Piel here at MME for ordering information).
The take away message: PLEASE...take a few minutes to become familiar with Koehler illumination and use it daily and check it whenever you move from one mag to another or one technique to another. Your microscopy will improve dramatically.
Best regards, 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 11:31 AM 4/4/2007, keith.morris-at-ucl.ac.uk wrote:
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Now, lets get one thing straight from the beginning. I use Koehler illumination. I think Koehler illumination is the mark of the competent microscopist. Don't use Koelhler illumination? As one of friends says "Dude, that's just wrong!"
But in truth the only scope I have true Koehler illumination is a monocular petrographic scope with a detached but focusable AO lamp with an iris. This scope is my own at home in my lab. All the scope I have seem and used in the last 20 years were missing some feature which prevented true Koehler illumination.
Some were lacking centerable lamps, others immovable ground glass filters while other did not have centerable objectives. Those that did had wire filaments and not ribbon filaments.
I don't care whose brand...It seems impossible to set up true classic Koehler illumination. (I don't even want to talk about focusable and centerable Bertrand lens!)
That my story and I'm sticking to it
Frank
==============================Original Headers============================== 7, 18 -- From frank.karl-at-degussa.com Wed Apr 4 11:55:59 2007 7, 18 -- Received: from framailout1.rz.itson.com (mailout2.degussa.com [149.216.91.173]) 7, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l34Gtv5T015417 7, 18 -- for {microscopy-at-msa.microscopy.com} ; Wed, 4 Apr 2007 11:55:58 -0500 7, 18 -- Received: from mobuscomm01.mail.degussa.com (uss1026.applications.degussanet.com [10.88.88.98]) 7, 18 -- by framailout1.rz.itson.com (8.13.4/8.13.4/Debian-3sarge3) with ESMTP id l34Gttw5022855 7, 18 -- for {microscopy-at-msa.microscopy.com} ; Wed, 4 Apr 2007 18:55:55 +0200 7, 18 -- In-Reply-To: {200704041623.l34GNX1E004458-at-ns.microscopy.com} 7, 18 -- Subject: Koehler illumination revisited 7, 18 -- To: microscopy-at-msa.microscopy.com 7, 18 -- X-Mailer: Lotus Notes Release 6.5.5 November 30, 2005 7, 18 -- Message-ID: {OF3967C455.9772638E-ON862572B3.005AA6D6-852572B3.005CF738-at-degussa.com} 7, 18 -- From: frank.karl-at-degussa.com 7, 18 -- Date: Wed, 4 Apr 2007 12:55:51 -0400 7, 18 -- X-MIMETrack: Serialize by Router on MOBUSComm01/DHexternal/US(Release 6.5.1|January 21, 2004) at 7, 18 -- 04/04/2007 11:55:56 AM 7, 18 -- MIME-Version: 1.0 7, 18 -- Content-type: text/plain; charset=US-ASCII ==============================End of - Headers==============================
This is more of a histology question than EM but is related. One of the labs I work for has had problems processing/staining sections on glass light microscopy slides when they use the Shandon Sequenza Coverplate technology* in a rack on a cooling plate in a microwave. The problem is small circular voids in the staining/labeling. This happens with both immuno and general tissue stains and is similar to a phenomena I occasionally see when staining semi-thin epoxy sections with toluidine blue on a hot plate. It appears as if there is some localized boiling resulting in a bubble of gas thus displacing the stain reagent.
My question is: have you experienced this phenomena and do you have a solution for eliminating it?
* This is a patented plastic device that fits over a standard microscope slide allowing specimens to be immunostained with a minimum of reagent (~ 80 microliters).
Thanks, Larry -- Larry Ackerman, Associate Specialist UCSF, Dept. of Anatomy, Rm S1347 513 Parnassus Ave., Box 0452 San Francisco, CA 94143
This has been an interesting thread to follow. Each of the replies has brought out some valid points, but I don't know if they quite got to Frank's questions.
It is important to consider nanometers (or microns) per pixel. That will determine the ultimate resolution you have to work with. Of course Nyquist will tell you that you can't push things to the single-pixel dimensions - a couple of pixels is more likely your limit.
It is also important to remember that raw pixel count alone is meaningless. You have to consider the image formation process. The camera needs to be matched to the phosphor for optimum cost and performance. Excess pixels in the camera beyond the resolution of the phosphor will just waste money. Insufficient pixels will forego potential resolution. I am not a TEM fellow, so I don't know the specifics of the camera systems, but I would like to think that systems are fairly well matched by the designers, at least now that the costs of CCDs are coming down. I also won't address the limits of microscope resolution. It will enter into the picture at some magnification.
Frank's questions seemed to deal with appreciating or visualizing the resolution once the image was captured.
On the computer screen, much software seems to display the images, or portions thereof, at one pixel of image per one pixel of screen. Many screens are setup so that pixels are NOT terribly obvious to the eye from normal viewing distance. Therefore, it will be difficult to notice one pixel more or less to a dimension without zooming in on the image. The software will have full access to the image data and can make measurements down to the pixel level.
The printed image also raises visualization issues. Multiple dots are required to render a single pixel, at least for those printers (laser and many inkjets) where a dot is either there or not. A pattern of dots is needed together to represent shades. Therefore, the printed pixels per inch is practically an order of magnitude less than the dots per inch. And then there are the "truth in labeling" issues. What is the printer genuinely capable of? Once again, the resolution of the eye comes into play. It is quoted at about 500 pixels (250 pixel pairs) per inch at 20 inches, but I don't think I would be appreciating one pixel more or less at that printed resolution. I have a hard time seeing jagginess in real-world, 1024-pixel-wide images printed at 5 inches. Zooming is necessary for me to clearly see individual pixels.
So it's time to get back to your original question about 3 megapixel cameras versus 1 megapixel cameras. My opinion is that you will only marginally appreciate the greater number of pixels on the screen or in a printed image. If your software maps images to the screen pixel by pixel, the image collected at a given magnification will be bigger on the screen and each pixel will represent a smaller dimension. The same software will probably print the image the same size and the question will be whether your eye will be able to appreciate the finer detail. It will offer larger prints (or more zoom) before pixel jagginess appears to the same degree.
For image analysis, the pixels will be finer at a given magnification (field of view). Therefore, you should be able to perform measurements on smaller features than before.
Warren Straszheim ---------------------------------------------------------------------------- The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
Let me float an question to the list. What is the effect of changing a one meg camera to a three meg camera on a TEM?
I'm asked some fundamental questions, and I can't seem to answer them to this person's satisfaction, which implies I don't have a good grasp of the question or answers.
My immediate response is you would increase the resolution. I can envision the image size on the monitor changing, but if the resolution of the screen is lower than the captured image and if the computer/imaging software wants to display all the image captured, will not any feature at a specific magnification have the resolution of the monitor? It seems the same is true for the printer. I can't simply expand the size of the paper at will, so the software will either reduce the printed image magnification or print just a smaller section of the total image. Again, since the printer has a fixed resolution will not the printed image resolution will be limited by the printer's (This sounds like a Hi-Fi discussion from the early 60's... just change the words...) upper limit?
So why capture high resolution images? My response is it allows you to post process and expand the image to examine one feature and have sufficient "stored" resolution to display the image without empty magnification. This also implies (to me at least) if I want to measure from point A to point B, the more camera pixels I have the better I can resolve where point A starts and point B ends which should allow me to have better confidence in my measurements. I believe that imaging software works on the image in memory and not the image displayed on the screen so size of the print or screen has little to do with data obtained. It's more a function of the size of the captured image?
Lastly.....
If I feel the need to have at least 1000 pixels in an image feature, and due to my camera I can only capture 500 at a magnification X, is there any reason not to simply increase the magnification so I have a larger feature which now occupies more pixels. I realize I haven't increased the resolution, but if my software need 999 pixels to "recognize" a feature haven't I met this requirement?
Thank in advance! Frank (I miss film) Karl
==============================Original Headers============================== 9, 30 -- From wesaia-at-iastate.edu Wed Apr 4 16:01:01 2007 9, 30 -- Received: from mailhub-3.iastate.edu (mailhub-3.iastate.edu [129.186.140.13]) 9, 30 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l34L11Rn014949 9, 30 -- for {Microscopy-at-msa.microscopy.com} ; Wed, 4 Apr 2007 16:01:01 -0500 9, 30 -- Received: from mailout-2.iastate.edu (mailout-2.iastate.edu [129.186.140.2]) 9, 30 -- by mailhub-3.iastate.edu (8.12.11.20060614/8.12.10) with SMTP id l34L11ad005490 9, 30 -- for {Microscopy-at-msa.microscopy.com} ; Wed, 4 Apr 2007 16:01:01 -0500 9, 30 -- Received: from (owa.eng.iastate.edu [129.186.23.85]) by mailout-2.iastate.edu with smtp 9, 30 -- id 10ec_7633dcb0_e2ef_11db_87e6_001372578af6; 9, 30 -- Wed, 04 Apr 2007 16:00:03 -0500 9, 30 -- Received: from maire.eng.iastate.edu ([10.10.196.69]) by owa.eng.iastate.edu with Microsoft SMTPSVC(6.0.3790.3959); 9, 30 -- Wed, 4 Apr 2007 16:01:01 -0500 9, 30 -- X-MIMEOLE: Produced By Microsoft Exchange V6.5 9, 30 -- Content-class: urn:content-classes:message 9, 30 -- MIME-Version: 1.0 9, 30 -- Content-Type: text/plain; 9, 30 -- charset="iso-8859-1" 9, 30 -- Subject: RE: [Microscopy] RE: Digital cameras and the TEM 9, 30 -- Date: Wed, 4 Apr 2007 16:01:14 -0500 9, 30 -- Message-ID: {16A330AC32056A40B32842EC4BB8D7270185BB5D-at-maire.eng.iastate.edu} 9, 30 -- X-MS-Has-Attach: 9, 30 -- X-MS-TNEF-Correlator: 9, 30 -- Thread-Topic: [Microscopy] RE: Digital cameras and the TEM 9, 30 -- Thread-Index: AcdypZGl86CNEaD0TGGwtRBUzX/9eABC/wKk 9, 30 -- References: {200703300829.l2U8Tepk002021-at-ns.microscopy.com} 9, 30 -- From: "Straszheim, Warren E [CCE E]" {wesaia-at-iastate.edu} 9, 30 -- To: "MSA" {Microscopy-at-msa.microscopy.com} 9, 30 -- X-OriginalArrivalTime: 04 Apr 2007 21:01:01.0501 (UTC) FILETIME=[59DF96D0:01C776FC] 9, 30 -- Content-Transfer-Encoding: 8bit 9, 30 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l34L11Rn014949 ==============================End of - Headers==============================
There may be two items happening. First is incomplete covering of the sections by the reagents. I am assuming this is a capillary action type of slide holder/stainer. Second is exactly what you mentioned, micro-boiling of reagent.
Possible ways to help with the first is to add a surfactant/detergent to reagents, i.e. triton/saponin in very dilute amounts.
Possible way to help with second is to lower the wattage of the microwave, due multiple short times of microwaving, add more heat sinks to oven or go back to the old tried and true method of overnight incubations at 4 degrees.
Ed
Edward P. Calomeni Director EM Lab - Pathology The Ohio State University M018 Starling Loving Hall 320 W. 10th Ave. Columbus, OH 43210 614-293-5580 (office) 614-293-8806 (lab) edward.calomeni-at-osumc.edu -----Original Message----- X-from: larry.ackerman-at-ucsf.edu [mailto:larry.ackerman-at-ucsf.edu] Sent: Wednesday, April 04, 2007 1:29 PM To: Calomeni, Edward
This is more of a histology question than EM but is related. One of the labs I work for has had problems processing/staining sections on glass light microscopy slides when they use the Shandon Sequenza Coverplate technology* in a rack on a cooling plate in a microwave. The problem is small circular voids in the staining/labeling. This happens with both immuno and general tissue stains and is similar to a phenomena I occasionally see when staining semi-thin epoxy sections with toluidine blue on a hot plate. It appears as if there is some localized boiling resulting in a bubble of gas thus displacing the stain reagent.
My question is: have you experienced this phenomena and do you have a solution for eliminating it?
* This is a patented plastic device that fits over a standard microscope slide allowing specimens to be immunostained with a minimum of reagent (~ 80 microliters).
Thanks, Larry -- Larry Ackerman, Associate Specialist UCSF, Dept. of Anatomy, Rm S1347 513 Parnassus Ave., Box 0452 San Francisco, CA 94143
Larry, we use this system and have not had this problem. However, in another lab, in another galaxy, far, far away, our IHC lab had problems with this. We deduced it was air bubble formation, and reduced the concentration of detergent used in the rinse buffer. We currently use 0.05% Tween 20 and have no problems with it.
Careful application of the slides to the coverplate is also warranted! If you wish, you can contact me off list and I'll give you our procedure for mounting the slides.
Hope this helps!
Teri Johnson, HT(ASCP)QIHC Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, MO 64110
-----Original Message----- X-from: larry.ackerman-at-ucsf.edu [mailto:larry.ackerman-at-ucsf.edu] Sent: Wednesday, April 04, 2007 12:31 PM To: Johnson, Teri
This is more of a histology question than EM but is related. One of the labs I work for has had problems processing/staining sections on glass light microscopy slides when they use the Shandon Sequenza Coverplate technology* in a rack on a cooling plate in a microwave. The problem is small circular voids in the staining/labeling. This happens with both immuno and general tissue stains and is similar to a phenomena I occasionally see when staining semi-thin epoxy sections with toluidine blue on a hot plate. It appears as if there is some localized boiling resulting in a bubble of gas thus displacing the stain reagent.
My question is: have you experienced this phenomena and do you have a solution for eliminating it?
* This is a patented plastic device that fits over a standard microscope
slide allowing specimens to be immunostained with a minimum of reagent (~ 80 microliters).
Thanks, Larry -- Larry Ackerman, Associate Specialist UCSF, Dept. of Anatomy, Rm S1347 513 Parnassus Ave., Box 0452 San Francisco, CA 94143
Ok. First of all, before I get yelled at for dismantling the column and stage of the SEM when removing it from the truck, be aware that the entire top portion of the console had shifted so badly during transit that the center of gravity was so far off that it was going to tip over if I even looked at it funny. The only way I saw to get it out of the truck without a major catastrophe was to take the column apart. Since the vacuum system was already pretty much dismantled, I didn't see that this would cause too much more pain.
As far as dismantling the control console goes, I did it because I had to get the transformer oil out of it. It was very slippery, and getting it clean was priority one. Thanks to those who informed me of the PCBs. I was aware of that, and I am taking as much precausion as possible, given the extent of the leakage. Again, not to fan the fires, I know that it is in the cafeteria, but it was pretty much contained in the control console and cleaned out before any parts were placed in the cafeteria. The table that the transformer is sitting on was broken and about to be thrown out anyway, so I used it so I don't have to heft the transformer off the floor.
So, for your perusal, here are the first sets of photos to come out of the restoration.
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 ewing.hr-at-bruker-axs.com as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Title-Subject: [Filtered] Job Opportunity at Bruker AXS
Question: Ý Microanalysis Applications Engineer
Bruker AXS, Inc., a leading manufacturer of Analytical Instrumentation, has a job opening for an Applications Engineer in their Ewing, NJ Microanalysis lab. This division specializes in Energy Dispersive X-ray (EDX) Analysis instrumentation for electron microscopes.
The position entails customer, service, sales and marketing support.
Primary Responsibilities: * Demonstration of the equipment at Bruker and customer sites * Analysis of customer samples and production of clear analysis reports * Provide technical support to existing, and potential customers via e- mail and phone * User training in the operation of the Bruker Microanalysis System at Bruker and customer sites * Produce and maintain appropriate materials for customer training * Preparation of technical sales materials * Assist field service in interpreting and resolving customer problems * Perform other tasks as assigned by manager or supervisor.
This position involves up to 30% travel, primarily in North America.
Candidates interested in this position require a minimum of an Associates degree in a Physical Science discipline with a Bachelors degree in Chemistry, Geology or Materials Science preferred. Two years SEM/X-ray microanalysis experience, including sample preparation, is preferred.
Requirements include excellent communication skills and proficiency with Microsoft Office.
Send resume and salary requirement to:
Ted Juzwak Bruker AXS Microanalysis, Inc. 1239 Parkway Avenue, Suite 203 Ewing, NJ 08628
ewing.hr-at-bruker-axs.com
Bruker AXS Inc. offers a competitive salary and comprehensive benefits package including health and dental insurance, Flexible Spending Account, company sponsored life and disability insurance, 401(k) plan with company matching components, and vacation and sick/personal days.
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 ewing.hr-at-bruker-axs.com as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Title-Subject: [Filtered] Job Opportunity at Bruker AXS
Question: * Microanalysis Applications Engineer
Bruker AXS, Inc., a leading manufacturer of Analytical Instrumentation, has a job opening for an Applications Engineer in their Ewing, NJ Microanalysis lab. This division specializes in Energy Dispersive X-ray (EDX) Analysis instrumentation for electron microscopes.
The position entails customer, service, sales and marketing support.
Primary Responsibilities: * Demonstration of the equipment at Bruker and customer sites * Analysis of customer samples and production of clear analysis reports * Provide technical support to existing, and potential customers via e- mail and phone * User training in the operation of the Bruker Microanalysis System at Bruker and customer sites * Produce and maintain appropriate materials for customer training * Preparation of technical sales materials * Assist field service in interpreting and resolving customer problems * Perform other tasks as assigned by manager or supervisor.
This position involves up to 30% travel, primarily in North America.
Candidates interested in this position require a minimum of an Associates degree in a Physical Science discipline with a Bachelors degree in Chemistry, Geology or Materials Science preferred. Two years SEM/X-ray microanalysis experience, including sample preparation, is preferred.
Requirements include excellent communication skills and proficiency with Microsoft Office.
Send resume and salary requirement to:
Ted Juzwak Bruker AXS Microanalysis, Inc. 1239 Parkway Avenue, Suite 203 Ewing, NJ 08628
ewing.hr-at-bruker-axs.com
Bruker AXS Inc. offers a competitive salary and comprehensive benefits package including health and dental insurance, Flexible Spending Account, company sponsored life and disability insurance, 401(k) plan with company matching components, and vacation and sick/personal days.
Ok. Today we concentrated on getting the column put back together. So far so good. We got to the point where we're doing preliminary vacuum checks- seeing if it holds a vacuum without powering it on, but manually actuating the valves. It doesn't. After about an hour and a half under the column checking every gasket, we discovered a gaping hole where there was at one point an EDS system. Ok.
We need the blanking cover for the angled opening that heads down into the chamber! If anybody has one lying around they aren't using, I would greatly appreciate it!
We are also missing the little thingy that one uses to place the specimen in the chamber through the airlock. I'm staring at an open hole, and I know that there is some kind of rod that pushes the specimen in, but we don't have it. If there is an extra somewhere, I'd appreciate it. If you even have one that you could photograph closely for me so I can see if I can make one, I would appreciate that as well.
I've posted more pictures on my site- http://www.jkraft.net/semproject
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Question: I use a Zeiss Confocor 2 to look at diffusion of GFP in yeast cells. My plot of G(t) sometimes contains a very strong sine wave-like oscillation with a period of about 100 hertz, and it ruins the curve. There is no obvious oscillation in the raw fluorescence intensity plot.
We guess that there is some sort of mechanical vibration in the room, but the scope is on an air table. The yeast cells are physically stuck to my slide and don't move at all. I don't see the oscillation if I look at fluorophores in solution.
Does anyone have any idea what this is and how to eliminate it?
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Email: roberta. burns-at-ametek.com Name: Roberta Burns
Organization: EDAX
Title-Subject: [Filtered] Job Opportunity at EDAX
Question: EDAX, Inc., a leading mnaufacturer of X-Ray Microanalysis Equipment has an opening for a Technical Product Manager for TEM Products. The position will handle all matters pertaining to EDAX elemental analysis and crystalography products for the TEM to include stategic product planning, product specification, budget/financial planning, marketing programs and sales support.
The position requires a Masters Degree in Science/Engineering and 5 years of relevant work experience.
Plesas send resumes to: Roberta Burns Human Resources Manager EDAX Inc. 91 McKee Drive Mahwah, NJ 07430
Time-Lapse Microscopy Short Course Live Cell Imaging for Biomedical Applications
2-4 May 2007, Villa Orlandi (Capri)
The University of Napoli “Federico II”, with the patronage of the Italian Cell Culture Association, is organizing a time-lapse microscopy course on May 2–4, 2007 in Capri. The course is addressed to both senior scientists and students interested in live cell imaging techniques. Both lectures and practical sessions on video microscopy workstations will be provided.
Course organizers Stefano Guido and Marino Simeone (University of Napoli “Federico II”, Italy)
Course overview The recent advances in microscopy techniques, coupled with the development of fluorescence markers, have provided scientists with an array of experimental tools to follow processes at the cellular level in a dynamic fashion. The purpose of this course is to provide an intensive training on state-of-the-art methods for live cell imaging by optical microscopy, with special emphasis on time-lapse techniques. Starting with a basic introduction to optical microscopy, the topics covered in the course program include microscope incubation, 3D fluorescence and multidimensional microscopy, computer-controlled sample scanning and optical sectioning, image processing and archiving, cell tracking, specialized fluorescence techniques (FRAP, FLIP, FRET, SPIM, TIRFM), and time-lapse applications.
Invited speakers: - Alberto Diaspro (Università di Genova, Italy) - Ernst H. K. Stelzer (EMBL, Heidelberg, Germany) - Peter Evennett (Royal Microscopical Society, UK) - Spencer L. Shorte (Institut Pasteur, Paris, France) - M. Cristina Cardoso (Max Delbrück Center for Molecular Medicine MDC, Berlin, Germany) - Roman S. Polishchuk (Consorzio Mario Negri Sud, Chieti, Italy) - Elena V. Polishchuk (Consorzio Mario Negri Sud, Chieti, Italy) - Tony Collins (McMaster University, Hamilton, Canada) - Dario Parazzoli (National Institute of Molecular Genetics INGM, Milano, Italy)
MAIN SPONSOR: Okolab, Italy (http://www.oko-lab.com) OTHER SPONSORS (to be updated): Zeiss, Hamamatsu, Coherent, Molecular Cytomics Official media partner: Imaging & Microscopy.
Please find more information on the course website: http://www.time-lapse-microscopy.unina.it or send an email to segreteria-at-time-lapse-microscopy.unina.it
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==============================Original Headers============================== 12, 24 -- From steguido-at-unina.it Fri Apr 6 08:52:35 2007 12, 24 -- Received: from webmail.unina.it (webmail.unina.it [192.132.34.212]) 12, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l36DqZjc011450 12, 24 -- for {Microscopy-at-microscopy.com} ; Fri, 6 Apr 2007 08:52:35 -0500 12, 24 -- Received: from webmail.unina.it (localhost [127.0.0.1]) 12, 24 -- by webmail.unina.it (8.12.11/8.12.11) with ESMTP id l36DwbwW004162 12, 24 -- for {Microscopy-at-microscopy.com} ; Fri, 6 Apr 2007 15:58:37 +0200 12, 24 -- Received: (from nobody-at-localhost) 12, 24 -- by webmail.unina.it (8.12.11/8.12.11/Submit) id l36Dwb7m004161 12, 24 -- for Microscopy-at-microscopy.com; Fri, 6 Apr 2007 15:58:37 +0200 12, 24 -- X-Authentication-Warning: webmail.unina.it: nobody set sender to steguido-at-unina.it using -f 12, 24 -- Received: from ichim83.ingchim.unina.it (ichim83.ingchim.unina.it [143.225.238.83]) 12, 24 -- by webmail.unina.it (IMP) with HTTP 12, 24 -- for {steguido-at-192.132.34.41} ; Fri, 6 Apr 2007 15:58:37 +0200 12, 24 -- Message-ID: {1175867917.4616520d67415-at-webmail.unina.it} 12, 24 -- Date: Fri, 6 Apr 2007 15:58:37 +0200 12, 24 -- From: steguido-at-unina.it 12, 24 -- To: Microscopy-at-microscopy.com 12, 24 -- Subject: LM - Time-lapse microscopy course announcement 12, 24 -- MIME-Version: 1.0 12, 24 -- Content-Type: text/plain; charset=ISO-8859-1 12, 24 -- Content-Transfer-Encoding: 8bit 12, 24 -- User-Agent: Internet Messaging Program (IMP) 3.2.2 12, 24 -- X-Originating-IP: 143.225.238.83 ==============================End of - Headers==============================
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Email: jmastrangelo-at-ulbi.com Name: Joe Mastrangelo
Organization: Ultralife Batteries
Title-Subject: [Filtered] SEM preparation for insects, other recommendations
Question: Our company typically sponsors a tour for "National Bring your Child to Work Day". Now that we have a SEM in house, I hope to be able to make their visit to our lab a memorable one.
Since most of my imaging is done on powders, mixtures, coatings, etc. I figure something a little more interesting might be in order. I would like to appeal to the listers out there for suggestions that would have relatively simple preparation techniques. My initial thoughts go to spider "face", fly eyes/legs, etc.
My questions are:
1) Any suggestions besides spider "face", fly eye/leg that would really interest the kids and might be readily available for me to find? (please include preparation techniques)
2) What type of sample preparation should I use to dehydrate insects but maintain the structures I want to image? (I would hate to have their innards all over the inside of the chamber) I seem to remember some threads talking about dehydration with methanol?
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Email: dmayes-at-carilion.com Name: daniel mayes
Organization: carilion clinical laboratories
Title-Subject: [Filtered] technical TEM services for renal biopsy service
Question: We are a laboratory serving the Carilion Health System, a network of hospitals and healthcare facilities located mainly in southwestern Virgina, and have a renal biopsy volume requiring electron microscopy of about one per month. We are looking for a facility to perform the technical service of TEM for these renal biopsies. The interpretation of the formalin fixed material and the immunofluorescence studies are performed here. The interpretation of the electron microscopic images could be performed either by us or by the performing facility. Please contact me at dmayes-at-carilion.com, or at 540-981-7889.
Joe, My experience has been that small, hard-bodied insects like small spiders, ants, ticks, ladybugs, even bumblebees, can just be mounted on a stub and sputtered. In fact fresh killed, or even live insects, don't even need to be coated until they are fully dessicated. Small spiders are particularly tough and may just walk away after an hour or 2 under vacuum! Pieces of leaf (especially the stoma on the bottom side) or very small flowers can be interesting with just sputtering. The more recognizable the object on the screen, the better the impact.
I'm assuming this is not an FESEM. I might be a little less callous about the vacuum if it were.
If you have TV scan rate available and can find an old wind-up watch, take the back off and watch it run.
Ken Converse owner
QUALITY IMAGES Servicing Scanning Electron Microscopes Since 1981 474 So. Bridgton Rd. Bridgton, ME 04009 207-647-4348 Fax 207-647-2688 kenconverse-at-qualityimages.biz qualityimages.biz
-----Original Message----- X-from: jmastrangelo-at-ulbi.com [mailto:jmastrangelo-at-ulbi.com] Sent: Friday, April 06, 2007 9:56 PM To: kenconverse-at-qualityimages.biz
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please copy both jmastrangelo-at-ulbi.com as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: jmastrangelo-at-ulbi.com Name: Joe Mastrangelo
Organization: Ultralife Batteries
Title-Subject: [Filtered] SEM preparation for insects, other recommendations
Question: Our company typically sponsors a tour for "National Bring your Child to Work Day". Now that we have a SEM in house, I hope to be able to make their visit to our lab a memorable one.
Since most of my imaging is done on powders, mixtures, coatings, etc. I figure something a little more interesting might be in order. I would like to appeal to the listers out there for suggestions that would have relatively simple preparation techniques. My initial thoughts go to spider "face", fly eyes/legs, etc.
My questions are:
1) Any suggestions besides spider "face", fly eye/leg that would really interest the kids and might be readily available for me to find? (please include preparation techniques)
2) What type of sample preparation should I use to dehydrate insects but maintain the structures I want to image? (I would hate to have their innards all over the inside of the chamber) I seem to remember some threads talking about dehydration with methanol?
==============================Original Headers============================== 13, 11 -- From zaluzec-at-microscopy.com Fri Apr 6 20:51:53 2007 13, 11 -- Received: from [206.69.208.22] (mac22.zaluzec.com [206.69.208.22]) 13, 11 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l371pq8P005220 13, 11 -- for {microscopy-at-microscopy.com} ; Fri, 6 Apr 2007 20:51:53 -0500 13, 11 -- Mime-Version: 1.0 13, 11 -- Message-Id: {p06240800c23ca9ae0f8b-at-[206.69.208.22]} 13, 11 -- Date: Fri, 6 Apr 2007 20:51:52 -0500 13, 11 -- To: microscopy-at-microscopy.com 13, 11 -- From: jmastrangelo-at-ulbi.com (by way of MicroscopyListserver) 13, 11 -- Subject: viaWWW: SEM preparation for insects, other recommendations 13, 11 -- Content-Type: text/plain; charset="us-ascii" ==============================End of - Headers==============================
_________________________________________________________________ Need personalized email and website? Look no further. It's easy with Doteasy $0 Web Hosting! Learn more at www.doteasy.com
==============================Original Headers============================== 33, 29 -- From kenconverse-at-qualityimages.biz Sat Apr 7 11:10:58 2007 33, 29 -- Received: from dpmailmta03.doteasy.com (dpmailmta03.doteasy.com [65.61.209.80]) 33, 29 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l37GAsMD009829 33, 29 -- for {Microscopy-at-MSA.Microscopy.Com} ; Sat, 7 Apr 2007 11:10:56 -0500 33, 29 -- Received: from qualityimages.biz (unverified [192.168.101.16]) 33, 29 -- by dpmailmta02.doteasy.com (DEO) with ESMTP id 71564427-1814644 33, 29 -- for multiple; Sat, 07 Apr 2007 10:39:42 -0700 33, 29 -- Received: from Ken [76.179.237.214] by qualityimages.biz with ESMTP 33, 29 -- (SMTPD32-8.05) id A28B4B10136; Sat, 07 Apr 2007 09:10:51 -0700 33, 29 -- From: "Ken Converse" {kenconverse-at-qualityimages.biz} 33, 29 -- To: {jmastrangelo-at-ulbi.com} , "MSA Listserver" {Microscopy-at-MSA.Microscopy.Com} 33, 29 -- Subject: RE: [Microscopy] viaWWW: SEM preparation for insects, other recommendations 33, 29 -- Date: Sat, 7 Apr 2007 12:10:27 -0400 33, 29 -- Message-ID: {001201c7792f$448b4470$6401a8c0-at-Ken} 33, 29 -- MIME-Version: 1.0 33, 29 -- Content-Type: text/plain; 33, 29 -- charset="US-ASCII" 33, 29 -- X-Priority: 3 (Normal) 33, 29 -- X-MSMail-Priority: Normal 33, 29 -- X-Mailer: Microsoft Outlook, Build 10.0.6822 33, 29 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.3028 33, 29 -- Importance: Normal 33, 29 -- Thread-Index: Acd4t9iAGIU608maQMmP1ffGRdCcaAAdfJYg 33, 29 -- In-Reply-To: {200704070155.l371tXXK014506-at-ns.microscopy.com} 33, 29 -- X-IMSTrailer: __IMail_7__ 33, 29 -- X-IP-stats: Incoming Last 0, First 191, in=3975940, out=0, spam=0 ip=192.168.101.16 33, 29 -- X-Originating-IP: 192.168.101.16 33, 29 -- Content-Transfer-Encoding: 8bit 33, 29 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l37GAsMD009829 ==============================End of - Headers==============================
Hi Joe, this is an excellent oportunity to get kids excited about science and microscopy. I have done this several times when I was working as a scientist, and it's alwys fun.
I worked as a materials scientist, and we did not have any CPD equipment, so I simply looked for dead insects in the nooks and crannies of the lab and my home. Typically you find a good supply in corners of the basement windows. I simply glued them to a sample holder stub (you need to do this carefully, as they can be brittle and break. Then apply a thin coating of gold and you have a sample that you can put in the chamber. Put in a few. A fly, a spider, an ant, and a bug.
Perhaps a few pointers:
You can put in a sample of your powders, but the kids will most likely not be interested in the details of grain structure or fractal dimensions. They will lose interest quickly and become fidgety. Better stick to flys and bugs.
Before the fun starts, put everything in your lab away that is small or potentially dangerous. Depending on the size of the group, it is hard to control the kids in the dark. I usually had a tape that I used to block of an area where nobody was allowed to go, and everything that I could not put in a cabinet was behind the tape out of reach.
The kids seemed to like to be engaged. Instead of "zooming in" on the bug, I started out at high mag and had the kids guess at what they were seeing. Ask them to be specific, not just "a bug". An interesting starting point is the eye of an insect. You can zoom in between several segments (sometimes there are hair-like structures that stick out from there). Once you zoom out enough, the kids will identify it as a bug's eye. The first one to give me a good answer was then allowed to "run" the scope (change mag, and move the sample holder, slowly, of course).
Ask them what they would do if they had an SEM. Then use that to explain some of the features. One kid, for example, told me he would use it to watch the fish in his aquarium. Good hook to explain about vacuum, live tissue in vacuum, why everything is black and white, etc.
Most importantly: have fun with it yourself. The kids react to your engagement as much as they do to the pictures.
mike
Michael Bode
General Manager
OLYMPUS SOFT IMAGING SOLUTIONS CORP. 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.net www.olympus-sis.net
-----Original Message----- X-from: jmastrangelo-at-ulbi.com [mailto:jmastrangelo-at-ulbi.com] Sent: Friday, April 06, 2007 19:58 To: Mike Bode
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 jmastrangelo-at-ulbi.com as well as the MIcroscopy Listserver ------------------------------------------------------------------------ ---
Email: jmastrangelo-at-ulbi.com Name: Joe Mastrangelo
Organization: Ultralife Batteries
Title-Subject: [Filtered] SEM preparation for insects, other recommendations
Question: Our company typically sponsors a tour for "National Bring your Child to Work Day". Now that we have a SEM in house, I hope to be able to make their visit to our lab a memorable one.
Since most of my imaging is done on powders, mixtures, coatings, etc. I figure something a little more interesting might be in order. I would like to appeal to the listers out there for suggestions that would have relatively simple preparation techniques. My initial thoughts go to spider "face", fly eyes/legs, etc.
My questions are:
1) Any suggestions besides spider "face", fly eye/leg that would really interest the kids and might be readily available for me to find? (please include preparation techniques)
2) What type of sample preparation should I use to dehydrate insects but maintain the structures I want to image? (I would hate to have their innards all over the inside of the chamber) I seem to remember some threads talking about dehydration with methanol?
--| --| --|Hello, --| --|I am trying to simulate electron beam heating in the SEM. I am sure --|this is not a new topic and perhaps lots of people had done some --|work on it. I am totally new to this area so like to check if anyone --|has good journals to recommend? --| --|In my simulation, I input a figure for the probe current density --|(got from some journals), I inevitably gets melting... I am still --|trying to verify this. --| --|Can anyone point out to me a typical figure for probe size, current --|and perhaps even current distribution equation for a TEM probe? --| --|Thank you very much --| --|Regards --|TT
The current density may be high but the current itself is very low because you have essentially a point source in the specimen.
You are right, it has been done many times. Except for really good thermal insulators (Styrofoam?) the heating is negligible (|--1 deg C) for beam currents of 10 to the minus 10 amps or lower. See Scanning Electron Microscopy by Oliver Wells (1975).
Damage is usually due not to heating but to "radiation damage" cause by the fact that most of the energy is deposited in "lumps" of more than 20 eV each (i.e., large enough for one "lump" to break a covalent bond). This is large and complex topic and is the reason that it is VERY hard to make images showing better resolution than 3 nm of any covalently-bonded material (i.e., all biology). (See Electron Crystallography on the web and authors such as Robert Glaeser, Wah Chiu, and Ken Downing)
Cheers,
Jim Pawley
-- **************************************** Prof. James B. Pawley, Ph. 608-263-3147 Room 223, Zoology Research Building, FAX 608-262-9083 250 N. Mills St., Madison, WI, 53706 JBPAWLEY-at-WISC.EDU "A scientist is not one who can answer questions but one who can question answers." Theodore Schick Jr., Skeptical Enquirer, 21-2:39 "He who can get you to believe absurdities, can get you to commit atrocities." Voltaire.
==============================Original Headers============================== 7, 26 -- From jbpawley-at-wisc.edu Sat Apr 7 16:30:55 2007 7, 26 -- Received: from agogare.doit.wisc.edu (agogare.doit.wisc.edu [144.92.197.211]) 7, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l37LUsQh006083 7, 26 -- for {Microscopy-at-Microscopy.Com} ; Sat, 7 Apr 2007 16:30:54 -0500 7, 26 -- Received: from avs-daemon.smtpauth2.wiscmail.wisc.edu by 7, 26 -- smtpauth2.wiscmail.wisc.edu 7, 26 -- (Sun Java System Messaging Server 6.2-8.04 (built Feb 28 2007)) 7, 26 -- id {0JG500603D3IT600-at-smtpauth2.wiscmail.wisc.edu} for 7, 26 -- Microscopy-at-Microscopy.Com; Sat, 07 Apr 2007 16:30:54 -0500 (CDT) 7, 26 -- Received: from [172.16.1.43] ([76.210.66.44]) by smtpauth2.wiscmail.wisc.edu 7, 26 -- (Sun Java System Messaging Server 6.2-8.04 (built Feb 28 2007)) 7, 26 -- with ESMTPSA id {0JG5005TAD3FVH00-at-smtpauth2.wiscmail.wisc.edu} for 7, 26 -- Microscopy-at-Microscopy.Com; Sat, 07 Apr 2007 16:30:53 -0500 (CDT) 7, 26 -- Date: Sat, 07 Apr 2007 16:30:49 -0500 7, 26 -- From: James Pawley {jbpawley-at-wisc.edu} 7, 26 -- Subject: RE: Electron beam simulation 7, 26 -- X-Sender: jbpawley-at-wiscmail.wisc.edu 7, 26 -- To: "ListServer-at-MSA.Microscopy.Com" {Microscopy-at-Microscopy.Com} 7, 26 -- Message-id: {p06110402c23dbd58fb17-at-[172.16.1.43]} 7, 26 -- MIME-version: 1.0 7, 26 -- Content-type: text/plain; charset=us-ascii; format=flowed 7, 26 -- Content-transfer-encoding: 7BIT 7, 26 -- X-Spam-Report: AuthenticatedSender=yes, SenderIP=76.210.66.44 7, 26 -- X-Spam-PmxInfo: Server=avs-12, Version=5.3.0.289146, 7, 26 -- Antispam-Engine: 2.5.0.283055, Antispam-Data: 2007.4.7.141834, 7, 26 -- SenderIP=76.210.66.44 ==============================End of - Headers==============================
I regularly had K1-12 students tour my facility at UCD (now deconstructed by some very stupid and unethical faculty-oops, did I say that out loud?) Anyhow, I had a group of stubs I prepared for the kids to view and try to guess what they were seeing. I had a stub with both a piece of feather and a moth wing on it. I had some very small bones and teeth from a mouse (recovered from an owl pellet). I had a beautiful fly prep but it was much too complicated too duplicate so I suggest a fly wing. Most people are surprised that the wing is covered with small hairs. Flies and spiders usually collapse upon drying but beetles, wasps, etc. will do fine with air drying. As has been noted, many insects can be mounted live and survive the ordeal and many are even seen to move in the vacuum though usually they are immobilized by the expansion of the gases in their haemolymph. I would notify the class to bring in something to look at (within reason). Had to be no bigger than a penny, hard not soft, clean as possible. Sure, you will get scabs and buggers but you often get something worthwhile and the kids love that. We looked at mechanical watch parts, isopods (pill bugs), we compared hair from different animals (all on one stub), I had some nice cross-sections of wood, interesting leaves, insect eggs (Lepidoptera) with micropyle visible. Another good stub is an Argentine ant, the little tiny guys you get in your home, mounted on the same stub as a large ant like Pogonomyrmex or better yet, Camponotus, one of the carpenter ants. If you are careful, you can mount the tiny ant on top of the larger ant
I made a multi-stub holder and could arrange 6 stubs at once in our Hitachi S3500N. Always a big hit with the kids.
Rick Harris Informational and Educational Technology (no faculty in this division!!) University of California at Davis
==============================Original Headers============================== 5, 26 -- From raharris-at-ucdavis.edu Mon Apr 9 11:40:12 2007 5, 26 -- Received: from warsaw.ucdavis.edu (warsaw.ucdavis.edu [128.120.32.41]) 5, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l39GeCwB020009 5, 26 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 9 Apr 2007 11:40:12 -0500 5, 26 -- Received: from VEXBE2.ex.ad3.ucdavis.edu (exbe2.ucdavis.edu [169.237.229.69]) 5, 26 -- by warsaw.ucdavis.edu (8.13.7/8.13.1/it-defang-5.4.0) with ESMTP id l39GeAqr006098 5, 26 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 9 Apr 2007 09:40:11 -0700 (PDT) 5, 26 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 5, 26 -- Content-class: urn:content-classes:message 5, 26 -- MIME-Version: 1.0 5, 26 -- Content-Type: text/plain; 5, 26 -- charset="US-ASCII" 5, 26 -- Subject: RE: [Microscopy] RE: viaWWW: SEM preparation for insects, other recommendations 5, 26 -- Date: Mon, 9 Apr 2007 09:40:10 -0700 5, 26 -- Message-ID: {865D53A6C8279540A152822700036F6D0807E661-at-VEXBE2.ex.ad3.ucdavis.edu} 5, 26 -- In-Reply-To: {200704071617.l37GHdMq015735-at-ns.microscopy.com} 5, 26 -- X-MS-Has-Attach: 5, 26 -- X-MS-TNEF-Correlator: 5, 26 -- Thread-Topic: [Microscopy] RE: viaWWW: SEM preparation for insects, other recommendations 5, 26 -- Thread-Index: Acd5MEhIy8Z9NBbQT46pnoGpCynvpwBkSzhQ 5, 26 -- References: {200704071617.l37GHdMq015735-at-ns.microscopy.com} 5, 26 -- From: "Rick A Harris" {raharris-at-ucdavis.edu} 5, 26 -- To: {Microscopy-at-MSA.Microscopy.Com} 5, 26 -- X-Scanned-By: MIMEDefang 2.57 on 128.120.32.41 5, 26 -- Content-Transfer-Encoding: 8bit 5, 26 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l39GeCwB020009 ==============================End of - Headers==============================
Have any of you had success imaging higher plant microtubules? If so than I would really appreciate the method used.
I would also appreciate a fixation protocol for microbes, yeast, or animal microtubules that is particularly successful. This would be a starting point if we cannot find one used on higher plants.
Although high pressure freezing and FS is an option (protocol?) I would like to start with a chemical fixation as it would permit faster turn-around to possibly see if the experiment is sufficiently promising to go further.
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============================== 7, 21 -- From dsherman-at-purdue.edu Tue Apr 10 21:27:09 2007 7, 21 -- Received: from 1061exfe04.adpc.purdue.edu (1061exfe04.adpc.purdue.edu [128.210.63.227]) 7, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3B2R9Bt003164 7, 21 -- for {microscopy-at-microscopy.com} ; Tue, 10 Apr 2007 21:27:09 -0500 7, 21 -- Received: from exchange.purdue.edu ([172.21.6.23]) by 1061exfe04.adpc.purdue.edu with Microsoft SMTPSVC(6.0.3790.211); 7, 21 -- Tue, 10 Apr 2007 22:27:08 -0400 7, 21 -- Received: from 74.140.106.146 ([74.140.106.146]) 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 ; 7, 21 -- Wed, 11 Apr 2007 02:27:07 +0000 7, 21 -- User-Agent: Microsoft-Entourage/11.3.3.061214 7, 21 -- Date: Tue, 10 Apr 2007 22:27:08 -0400 7, 21 -- Subject: Plant microtubules 7, 21 -- From: Debby Sherman {dsherman-at-purdue.edu} 7, 21 -- To: "message to: MSA list" {microscopy-at-microscopy.com} 7, 21 -- Message-ID: {C241BFBC.BE2D%dsherman-at-purdue.edu} 7, 21 -- Thread-Topic: Plant microtubules 7, 21 -- Thread-Index: Acd74ObkJbPA1ufUEdulbAAKlcoUxg== 7, 21 -- Mime-version: 1.0 7, 21 -- Content-type: text/plain; 7, 21 -- charset="US-ASCII" 7, 21 -- Content-transfer-encoding: 7bit 7, 21 -- X-OriginalArrivalTime: 11 Apr 2007 02:27:08.0851 (UTC) FILETIME=[E766C030:01C77BE0] ==============================End of - Headers==============================
Ok. We've got it put together, but there are a couple of things that I'm dwelling on. First of all, there is an ion pump but no ion pump controller. Do I need the ion pump if I'm not running a LaB6 cathode?
Second, there are a couple of as-yet unidentified modules in the control console. These are labeled "MSI" "STB" and "SDU." Can anyone tell me what these modules are for? I can't make it out from following the connecting wires.
Finally, we have an optical microscope and WDS spectrometers on the instrument. I really don't have the ability to use these. I would love to do some X-ray spectroscopy, however we have no controller or software to use with these detectors.
2: Any type of digital image acquisition system. We have the photographic system, but we really can't afford the film for the number of students that will want pictures of everything that they see. We're using this with middle school students as well as high school students.
3: Specimen holders. Right now the stage just has the opening that goes all the way through the rotation gear. There is no holder, and no sample insertion rod, etc... Anything along those lines would be great.
4: Ion pump controller. (This is fairly low on the priority list, and I would probably be willing to part with the ion pump in exchange towards something on our wish-list.)
5: Better vacuum pump. We have a small vacuum pump that kind-of works, but doesn't work well. I would like to get a better two-stage pump that is actually made for SEM work, as opposed to our A/C serviceman's pump.
We have some extra items that we can sell/trade for anything you might have extras of. First, the WDS detectors. (I can supply detailed pictures and such if you want them) All I would ask extra for the WDS detectors are the blanking plates- if I take them off, I'd need the plates.
I also have a Denton Desk II Carbon accessory, as well as a spare Desk II glass cylinder for the sputter coater, (But no coater to use it with).
If we don't need the ion pump, or if we don't get a controller, I'd trade that too.
Also, if anybody has broken unusable parts in the line of detectors and such (I have a broken PMT) that they would be willing to part with, I would love to have some disassembled stuff to demonstrate the different parts of the SEM. For instance, I would love to cut a diffusion pump in half to show how that works and make the connection to how much more of a vacuum is needed than say, their vacuum cleaner at home.
And finally, I would like to take a moment to thank everyone on this list for their insightful and helpful comments, as well as your support of this project over the last couple of weeks. I really could not have done any of this without the wonderful advice and well-wishes from the members of this group.
This is to inform you and to encourage you to consider participating in the topical conference on In-Situ Electron Microscopy organized within the 54th AVS international symposium (http://www2.avs.org/call/2007/main.html) October 14-19, 2007, Seattle.
The topical conference call for papers is at
http://www2.avs.org/call/2007/ie.html
AVS provides a unique opportunity of electron microscopists to interact with a wide circle of scientists who study the dynamics of materials by in-situ techniques.
The confirmed invited speakers are:
U. Dahmen, Lawrence Berkeley National Laboratory J.M. Howe, University of Virginia B. Kabius, Argonne National Laboratory I. Robertson, University of Illinois, Urbana-Champaign R. Sharma, Arizona State University E. Stach, Purdue University S. Takeda, Osaka University, Japan, "In-situ Environmental TEM of the Nucleation and Growth of One-Dimensional Nanostructures" J.M. Zuo, University of Illinois, Urbana-Champaign
If you have suggestion for names of other invited speakers please contact the topical conference organizers:
Suneel Kodambaka (kodambaka-at-ucla.edu) Ivan Petrov (petrov-at-uiuc.edu)
==============================Original Headers============================== 14, 21 -- From petrov-at-mrl.uiuc.edu Wed Apr 11 17:06:02 2007 14, 21 -- Received: from mrlnt6.mrl.uiuc.edu (mrlnt6.mrl.uiuc.edu [130.126.101.9]) 14, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3BM62GA019997 14, 21 -- for {Microscopy-at-microscopy.com} ; Wed, 11 Apr 2007 17:06:02 -0500 14, 21 -- Content-class: urn:content-classes:message 14, 21 -- MIME-Version: 1.0 14, 21 -- Content-Type: text/plain; 14, 21 -- charset="us-ascii" 14, 21 -- X-MimeOLE: Produced By Microsoft Exchange V6.0.6603.0 14, 21 -- Subject: In-Situ Electron Microscopy at AVS54 14, 21 -- Date: Wed, 11 Apr 2007 17:06:02 -0500 14, 21 -- Message-ID: {9EADC1E53F9C70479BF6559370369114428FF6-at-mrlnt6.mrl.uiuc.edu} 14, 21 -- X-MS-Has-Attach: 14, 21 -- X-MS-TNEF-Correlator: 14, 21 -- Thread-Topic: In-Situ Electron Microscopy at AVS54 14, 21 -- Thread-Index: Acd8hZfUBVVUL62rTwmIU7zXkJno9Q== 14, 21 -- From: "Ivan Petrov" {petrov-at-mrl.uiuc.edu} 14, 21 -- To: {Microscopy-at-microscopy.com} 14, 21 -- Cc: {kodambaka-at-ucla.edu} 14, 21 -- Content-Transfer-Encoding: 8bit 14, 21 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l3BM62GA019997 ==============================End of - Headers==============================
-- A postdoctoral or junior staff position, with demonstrated experience in electron cryo-microscopy, is open in the laboratory of Dr. Timothy S. Baker at the University of California - San Diego in the Chemistry and Biochemistry Department.
Applications will be accepted from individuals who possess a degree in the Natural Sciences (Ph. D. for postdoctoral position only). The salary and level of the position will be commensurate with the candidate's experience and skills.
The research focuses on structure-function studies of macromolecular assemblies (primarily viruses) with cryo-TEM and 3D image reconstruction techniques. Highly-motivated candidates with significant experience in cryo-TEM and/or cryo-tomography, along with experience in image reconstruction and molecular image analysis, are encouraged to apply. See http://cryoem.ucsd.edu/facilities.shtm for a description of facilities. Image processing is performed in-house on a three-node Linux cluster housed at the San Diego Supercomputer Center, an organized research unit of UCSD. Additional computations are performed using the NSF supported TeraGrid resources.
San Diego is a vibrant, cosmopolitan city, known for its beautiful, year-round climate. The area includes about 70 miles of beaches, and water sports such as surfing, snorkeling, kayaking and fishing are all very popular. Winter alpine skiing and the deserts are within a couple of hours drive. The city is home to both the San Diego Chargers and the San Diego Padres. Mexico is close enough for an afternoon trip.
Qualified applicants should send a CV that identifies areas of expertise and interest, employment history, a publications list, and the names and contact information of three, professional references to Dr. Timothy S. Baker, Professor of Chemistry/Biochemistry _and Molecular Biology, _University of California, San Diego_, 9500 Gilman Drive MC-0378, La Jolla, California 92093-0378. Alternatively, CVs may be sent to tsb-at-ucsd.edu. See http://cryoem.ucsd.edu for more information about the laboratory.
==============================Original Headers============================== 6, 31 -- From nholson-at-ucsd.edu Wed Apr 11 19:15:25 2007 6, 31 -- Received: from outbound2.ucsd.edu (outbound2.ucsd.edu [132.239.1.206]) 6, 31 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3C0FOxI001247 6, 31 -- for {Microscopy-at-microscopy.com} ; Wed, 11 Apr 2007 19:15:25 -0500 6, 31 -- Received: from mailbox5.ucsd.edu (mailbox5.ucsd.edu [132.239.1.57]) 6, 31 -- by outbound2.ucsd.edu (8.13.6/8.13.6) with ESMTP id l3C0FN3i077619 6, 31 -- for {Microscopy-at-microscopy.com} ; Wed, 11 Apr 2007 17:15:24 -0700 (PDT) 6, 31 -- DomainKey-Signature: a=rsa-sha1; s=2007001; d=ucsd.edu; c=simple; q=dns; 6, 31 -- b=aZfWPRTdAui1fd0lhunPP1EhLJz8P8PHFXF+tDCe++NsSP3UNI+lSff+XYAIMtKg5 6, 31 -- gn3nWD4W2R3YDXv09Otyw== 6, 31 -- Received: from spameater.ucsd.edu (spameater.ucsd.edu [132.239.1.25]) 6, 31 -- by mailbox5.ucsd.edu (Postfix) with ESMTP id 74C0FNL00E8JW 6, 31 -- for {Microscopy-at-microscopy.com} ; Wed, 11 Apr 2007 17:15:23 -0700 (PDT) 6, 31 -- X-Virus-Scanned: amavisd-new at spameater.ucsd.edu 6, 31 -- Received: from mailbox5.ucsd.edu ([132.239.1.57]) 6, 31 -- by spameater.ucsd.edu (spameater.ucsd.edu [132.239.1.25]) (amavisd-new, port 10024) 6, 31 -- with ESMTP id L7M5q-UTezbu for {Microscopy-at-microscopy.com} ; 6, 31 -- Wed, 11 Apr 2007 17:15:23 -0700 (PDT) 6, 31 -- Received: from smtp.ucsd.edu (smtp.ucsd.edu [132.239.1.49]) 6, 31 -- by mailbox5.ucsd.edu (Postfix) with ESMTP id 74C0FNo00E8JR 6, 31 -- for {Microscopy-at-microscopy.com} ; Wed, 11 Apr 2007 17:15:23 -0700 (PDT) 6, 31 -- Received: from [132.239.70.186] (patrick-70-186.ucsd.edu [132.239.70.186]) 6, 31 -- by smtp.ucsd.edu (8.13.6/8.13.4) with ESMTP id l3C0FMXp081580 6, 31 -- for {Microscopy-at-microscopy.com} ; Wed, 11 Apr 2007 17:15:22 -0700 (PDT) 6, 31 -- Mime-Version: 1.0 6, 31 -- Message-Id: {p06230910c2432a675204-at-[132.239.70.186]} 6, 31 -- Date: Wed, 11 Apr 2007 17:15:11 -0700 6, 31 -- To: Microscopy-at-microscopy.com 6, 31 -- From: Norm Olson {nholson-at-ucsd.edu} 6, 31 -- Subject: Postdoc or JR Staff position at UC San Diego 6, 31 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" ==============================End of - Headers==============================
I have a Reichert Ultracut that is badly in need of a new motor drive belt. I have changed these belts on other microtomes, but not an Ultracut. It is not immediately obvious how the belt comes off the pully located behind the flywheel/handcrank. Does anyone have instructions for changing out this belt?
Who is the current technical representative for the older Reichert machines and does Leica Microsystems currently hold the Reichert name?
Thanks, Greg
-- -- ================================================================== Greg Strout Electron Microscopist, University of Oklahoma WWW Virtual Library for Microscopy: http://www.ou.edu/research/electron/www-vl/ e-mail: gstrout-at-ou.edu Opinions expressed herein are mine and not necessarily those of the University of Oklahoma ==================================================================
==============================Original Headers============================== 6, 19 -- From gstrout-at-ou.edu Thu Apr 12 12:05:47 2007 6, 19 -- Received: from ig88.ou.edu (mail.zero.ou.edu [129.15.0.75]) 6, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3CH5lrX003923 6, 19 -- for {microscopy-at-microscopy.com} ; Thu, 12 Apr 2007 12:05:47 -0500 6, 19 -- Received: from [127.0.0.1] ([129.15.38.202]) 6, 19 -- by ig88.ou.edu (Sun Java System Messaging Server 6.2-7.05 (built Sep 5 2006)) 6, 19 -- with ESMTP id {0JGE00KSKA1XOMK0-at-ig88.ou.edu} for microscopy-at-microscopy.com; 6, 19 -- Thu, 12 Apr 2007 12:05:34 -0500 (CDT) 6, 19 -- Date: Thu, 12 Apr 2007 12:03:28 -0500 6, 19 -- From: Greg Strout {gstrout-at-ou.edu} 6, 19 -- Subject: Reichert Ultracut motor drive belt 6, 19 -- To: microscopy-at-microscopy.com 6, 19 -- Message-id: {461E6660.7080601-at-ou.edu} 6, 19 -- MIME-version: 1.0 6, 19 -- Content-type: text/plain; charset=ISO-8859-1; format=flowed 6, 19 -- Content-transfer-encoding: 7BIT 6, 19 -- X-Accept-Language: en-us, en 6, 19 -- User-Agent: Mozilla/5.0 (Windows; U; Windows NT 5.1; en-US; rv:1.7.2) 6, 19 -- Gecko/20040804 Netscape/7.2 (ax) ==============================End of - Headers==============================
3rd UVM Practical Course on Three-dimensional Cryo Electron Microscopy of Single Particles August 13-19 2007, University of Vermont, Burlington, VT
This international course will teach the principles of three-dimensional reconstruction of single particles from cryo electron micrographs. The course will include a lecture series teaching in-depth the basic theoretical principles of single particle analysis and discussions and demonstrations of experimental procedures. Six hours per day are allocated for hands-on processing of data sets from electron micrographs of single particles.
Participants will work in groups of two. Each group will have one instructor, who will provide step-by-step guidance through the complete three-dimensional reconstruction process and offer additional explanations and support. The course aims to provide participants with a strong practical and theoretical background that will enable them later to use these techniques in their home laboratory.
For details please visit:
http://physioweb.med.uvm.edu/Cryo_Practical/
______________________________________________ Michael Radermacher, Assoc. Prof. University of Vermont Dept. Molecular Physiology and Biophysics HSRF 120 / 149 Beaumont Ave Burlington, VT 05405
==============================Original Headers============================== 15, 27 -- From mraderma-at-physiology.med.uvm.edu Thu Apr 12 13:36:43 2007 15, 27 -- Received: from pony.uvm.edu (pony.uvm.edu [132.198.101.202]) 15, 27 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3CIagRZ017433 15, 27 -- for {Microscopy-at-microscopy.com} ; Thu, 12 Apr 2007 13:36:42 -0500 15, 27 -- Received: from physiology.med.uvm.edu (physiology.med.uvm.edu [132.198.52.4]) 15, 27 -- by pony.uvm.edu (8.13.7/8.13.5) with ESMTP id l3CIadxH002961 15, 27 -- for {Microscopy-at-microscopy.com} ; Thu, 12 Apr 2007 14:36:39 -0400 15, 27 -- Received: from physiology.med.uvm.edu (localhost [127.0.0.1]) 15, 27 -- by physiology.med.uvm.edu (Postfix) with ESMTP id A03E8F326F 15, 27 -- for {Microscopy-at-microscopy.com} ; Thu, 12 Apr 2007 14:36:39 -0400 (EDT) 15, 27 -- Received: (from apache-at-localhost) 15, 27 -- by physiology.med.uvm.edu (8.12.8/8.12.8/Submit) id l3CIadxw024035 15, 27 -- for Microscopy-at-microscopy.com; Thu, 12 Apr 2007 14:36:39 -0400 15, 27 -- From: Michael Radermacher {mraderma-at-physiology.med.uvm.edu} 15, 27 -- X-Authentication-Warning: physiology.med.uvm.edu: apache set sender to mraderma-at-physiology.med.uvm.edu using -f 15, 27 -- Received: from 132.198.90.90 ( [132.198.90.90]) 15, 27 -- as user mraderma-at-localhost by physiology.med.uvm.edu with HTTP; 15, 27 -- Thu, 12 Apr 2007 14:36:39 -0400 15, 27 -- Message-ID: {1176402999.461e7c3730d73-at-physiology.med.uvm.edu} 15, 27 -- Date: Thu, 12 Apr 2007 14:36:39 -0400 15, 27 -- To: Microscopy-at-microscopy.com 15, 27 -- Subject: 3rd UVM course on 3D reconstruction of single particles 15, 27 -- MIME-Version: 1.0 15, 27 -- Content-Type: text/plain; charset=ISO-8859-1 15, 27 -- Content-Transfer-Encoding: 8bit 15, 27 -- User-Agent: Internet Messaging Program (IMP) 3.1 15, 27 -- X-Originating-IP: 132.198.90.90 ==============================End of - Headers==============================
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Email: wcrgs-at-aol.com Name: Ronald Obie
Organization: WCRG
Title-Subject: [Filtered] Potential difference across a microscope slide
Question: Hello. Does anyone know of a microscope slide available which has the capability for a potential difference to be placed across it? We would like to evaluate the charge of various particles by looking at the direction to which they move relative to the charge placed at the edge of a cover slip. Has anyone ever tried this? Are there references available? Thank you for your response. Ronald Obie WCRG 336-841-0264
Can anyone recommend a good reference for electron microscopy images of biological tissues and cells? I have Bozzola's Electron Microscopy: Principles and Techniques for Biologists, but would like a reference focused on identification of tissues/cells, rather than technique. Thanks and TGIF, Jessica Cervantes Bend Research, Inc Bend, OR
==============================Original Headers============================== 3, 14 -- From cervantes-at-bendres.com Fri Apr 13 14:55:46 2007 3, 14 -- Received: from BRIEX04A.bri.local (bc161112.bendcable.com [216.228.161.112]) 3, 14 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3DJtkcx012804 3, 14 -- for {Microscopy-at-microscopy.com} ; Fri, 13 Apr 2007 14:55:46 -0500 3, 14 -- MIME-Version: 1.0 3, 14 -- Content-Type: text/plain; 3, 14 -- charset="us-ascii" 3, 14 -- Subject: Reference for EM of Biological Tissues and Cells 3, 14 -- Date: Fri, 13 Apr 2007 12:55:45 -0700 3, 14 -- Message-ID: {8943D65F9AD70E4488AD6DE09F15088768C44C-at-BRIEX04A} 3, 14 -- From: "Cervantes, Jessica" {cervantes-at-bendres.com} 3, 14 -- To: {Microscopy-at-microscopy.com} 3, 14 -- Content-Transfer-Encoding: 8bit 3, 14 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l3DJtkcx012804 ==============================End of - Headers==============================
Due to some bureaucratic mumbo-jumbo, I am not able to use Uranyl Acetate (nuclear warheads, blah, blah, blah). I would like to stain some osmium-fixed tissue thin-sections for TEM; the procedure I'm following has a UA/Sato's lead stain step for the sections. Does anyone know of a suitable alternative? I did a quick google and listserver archive search and didn't find anything, but I'm hoping someone has run into this problem before and can suggest something.
Crossing my fingers, Jessica Cervantes Bend Research, Inc Bend, OR
==============================Original Headers============================== 3, 16 -- From cervantes-at-bendres.com Fri Apr 13 15:12:47 2007 3, 16 -- Received: from BRIEX04A.bri.local (bc161112.bendcable.com [216.228.161.112]) 3, 16 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3DKClG6024466 3, 16 -- for {Microscopy-at-microscopy.com} ; Fri, 13 Apr 2007 15:12:47 -0500 3, 16 -- MIME-Version: 1.0 3, 16 -- Content-Type: text/plain; 3, 16 -- charset="us-ascii" 3, 16 -- Subject: TEM: Alternative to Uranyl Acetate Stain 3, 16 -- Date: Fri, 13 Apr 2007 13:12:46 -0700 3, 16 -- Message-ID: {8943D65F9AD70E4488AD6DE09F15088768C44D-at-BRIEX04A} 3, 16 -- In-Reply-To: {8943D65F9AD70E4488AD6DE09F15088768C44C-at-BRIEX04A} 3, 16 -- References: {8943D65F9AD70E4488AD6DE09F15088768C44C-at-BRIEX04A} 3, 16 -- From: "Cervantes, Jessica" {cervantes-at-bendres.com} 3, 16 -- To: {Microscopy-at-microscopy.com} 3, 16 -- Content-Transfer-Encoding: 8bit 3, 16 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l3DKClG6024466 ==============================End of - Headers==============================
Dear Jessica, If you can find it, you can't do much better than HISTOLOGY: A Text and Atlas by Johannes AG Rhodin. The copy I have is copyrighted 1974. It was published by Oxford University Press. Lee -- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy & Histology Core Facility Manager, Optical Microscopy Core Facility Weill-Cornell Medical College
Histology: A text and Atlas was updated 2006 (paperback) with CD-ROM and is available from Barnes & Noble. Another excellent choice is Color Atlas of Cytology, Histology and Microscopic Anatomy by Wolfgang Kuehnel, also available from B&N. The paperback edition is excellent!
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: lcgould-at-med.cornell.edu [mailto:lcgould-at-med.cornell.edu] Sent: Friday, April 13, 2007 4:24 PM To: Bobrowski, Walter
Dear Jessica, If you can find it, you can't do much better than HISTOLOGY: A Text and Atlas by Johannes AG Rhodin. The copy I have is copyrighted 1974. It was published by Oxford University Press. Lee -- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy & Histology Core Facility Manager, Optical Microscopy Core Facility Weill-Cornell Medical College
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==============================Original Headers============================== 16, 31 -- From Walter.Bobrowski-at-pfizer.com Fri Apr 13 15:33:10 2007 16, 31 -- Received: from gromsgom01.pfizer.com (gromsgo.pfizer.com [148.168.224.84]) 16, 31 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3DKXAUZ015267 16, 31 -- for {microscopy-at-microscopy.com} ; Fri, 13 Apr 2007 15:33:10 -0500 16, 31 -- Received: from groamrexc02.amer.pfizer.com (groamrexc02.amer.pfizer.com [172.30.8.169]) 16, 31 -- by gromsgom01.pfizer.com (8.13.7/8.13.7) with ESMTP id l3DKX2dc027460 16, 31 -- for {microscopy-at-microscopy.com} ; Fri, 13 Apr 2007 16:33:10 -0400 16, 31 -- Received: from mopamrexc01.amer.pfizer.com ([170.116.32.254]) by groamrexc02.amer.pfizer.com with Microsoft SMTPSVC(6.0.3790.1830); 16, 31 -- Fri, 13 Apr 2007 16:33:09 -0400 16, 31 -- Received: from anaamrexm01.amer.pfizer.com ([162.48.101.10]) by mopamrexc01.amer.pfizer.com with Microsoft SMTPSVC(6.0.3790.1830); 16, 31 -- Fri, 13 Apr 2007 16:33:08 -0400 16, 31 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 16, 31 -- Content-class: urn:content-classes:message 16, 31 -- MIME-Version: 1.0 16, 31 -- Content-Type: text/plain; 16, 31 -- charset="us-ascii" 16, 31 -- Subject: Re: Reference for EM of Biological Tissues and Cells 16, 31 -- Date: Fri, 13 Apr 2007 16:33:09 -0400 16, 31 -- Message-ID: {CF89AF6ABD93B746A55BCCD9480A4B3D08F0BC8D-at-anaamrexm01.amer.pfizer.com} 16, 31 -- In-Reply-To: {200704132024.l3DKOFSd012554-at-ns.microscopy.com} 16, 31 -- X-MS-Has-Attach: 16, 31 -- X-MS-TNEF-Correlator: 16, 31 -- Thread-Topic: Re: Reference for EM of Biological Tissues and Cells 16, 31 -- thread-index: Acd+CbWhOMfUGz44SrSmvudamkHNqgAAMaoA 16, 31 -- From: "Bobrowski, Walter" {Walter.Bobrowski-at-pfizer.com} 16, 31 -- To: {microscopy-at-microscopy.com} 16, 31 -- X-OriginalArrivalTime: 13 Apr 2007 20:33:08.0801 (UTC) FILETIME=[F2959310:01C77E0A] 16, 31 -- X-Proofpoint-Virus-Version: vendor=fsecure engine=4.65.5502:2.3.11,1.2.37,4.0.164 definitions=2007-04-13_05:2007-04-13,2007-04-13,2007-04-13 signatures=0 16, 31 -- X-Proofpoint-Spam-Reason: safe 16, 31 -- Content-Transfer-Encoding: 8bit 16, 31 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l3DKXAUZ015267 ==============================End of - Headers==============================
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Email: neerajg-at-clemson.edu Name: Neeraj V. Gohad
Organization: Clemson University
Title-Subject: [Filtered] Animations for Tomography
Question: Dear all,
I will be giving a seminar on Electron Tomography in our department as a part of our departmental seminar series. I am searching for animations illustrating the process of tomography, I do have schematics and animations showing the weighted backprojections and aligned stacks. I am looking for an animations which shows the electrons hitting the specimen in the column as the tilt series is being taken. Does any one know a source where I can find an animation and or schematic?
As always I appreciate everyoneís help ,
Regards,
Raj.
Neeraj V. Gohad Graduate Research Assistant Department of Biological Sciences Clemson University, Clemson, SC-29634
Don Fawcett's The Cell (publisher = Saunders) has superb TEM views of most organelles. don't know if it is still in print.
At 02:57 PM 04/13/07, you wrote:
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Does anyone have a service manual for Ultracuts and /or Ultracut Es ?
Thanks Sally
Dr SJ Stowe Facility Coordinator ANU Electron Microscopy Unit ANU CRICOS#00120C
} -----Original Message----- } From: gstrout-at-ou.edu [mailto:gstrout-at-ou.edu] } Sent: Friday, 13 April 2007 3:06 AM } To: sally.stowe-at-anu.edu.au } Subject: [Microscopy] Reichert Ultracut motor drive belt } } } } } } --------------------------------------------------------------- } ------------- } The Microscopy ListServer -- CoSponsor: The Microscopy } Society of America To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver
==============================Original Headers============================== 7, 26 -- From sally.stowe-at-anu.edu.au Sun Apr 15 18:25:38 2007 7, 26 -- Received: from mail.rsbs.anu.edu.au (mail.rsbs.anu.edu.au [150.203.72.70]) 7, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3FNPb3T030338 7, 26 -- for {microscopy-at-microscopy.com} ; Sun, 15 Apr 2007 18:25:37 -0500 7, 26 -- Received: from rsbs2262 (rsbs2262.rsbs.anu.edu.au [150.203.36.144]) 7, 26 -- by mail.rsbs.anu.edu.au (Postfix) with ESMTP 7, 26 -- id D3E9EF44CF6; Mon, 16 Apr 2007 09:25:34 +1000 (EST) 7, 26 -- Reply-To: {sally.stowe-at-anu.edu.au} 7, 26 -- From: "Sally Stowe" {sally.stowe-at-anu.edu.au} 7, 26 -- To: {gstrout-at-ou.edu} , {microscopy-at-microscopy.com} 7, 26 -- Subject: RE: [Microscopy] Reichert Ultracut motor drive belt 7, 26 -- Date: Mon, 16 Apr 2007 09:25:34 +1000 7, 26 -- Message-ID: {002201c77fb5$5e27e730$9024cb96-at-rsbs.anu.edu.au} 7, 26 -- MIME-Version: 1.0 7, 26 -- Content-Type: text/plain; 7, 26 -- charset="us-ascii" 7, 26 -- Content-Transfer-Encoding: 7bit 7, 26 -- X-Priority: 3 (Normal) 7, 26 -- X-MSMail-Priority: Normal 7, 26 -- X-Mailer: Microsoft Outlook, Build 10.0.6626 7, 26 -- In-Reply-To: {200704121706.l3CH60Zb004083-at-ns.microscopy.com} 7, 26 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2800.1506 7, 26 -- Importance: Normal 7, 26 -- X-RSBS-MailScanner-Information: Please contact the ISP for more information 7, 26 -- X-RSBS-MailScanner: Found to be clean 7, 26 -- X-MailScanner-From: sally.stowe-at-anu.edu.au ==============================End of - Headers==============================
Ronald Obie wrote: ============================================================== Does anyone know of a microscope slide available which has the capability for a potential difference to be placed across it? We would like to evaluate the charge of various particles by looking at the direction to which they move relative to the charge placed at the edge of a cover slip. Has anyone ever tried this? Are there references available? ======================================================= We at SPI Supplies have offered a line of ITO (indium tin oxide) coated microscope slides, see URL http://www.2spi.com/catalog/standards/ITO-coated-slides-resistivities.html
If you want microscope slide sizes substrates, then at the bottom, click on "Microscopy Slides". They are also available with busbars which makes for an easier connection for electrical contacts. The slides are available at different resistivities.
Note that we also offer ITO coated cover slips, either with or without busbars.
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============================== 9, 26 -- From cgarber-at-2spi.com Sun Apr 15 22:25:09 2007 9, 26 -- Received: from diskless5.axs2000.net (mail.axs2000.net [209.120.196.43]) 9, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3G3P8J2013042 9, 26 -- for {microscopy-at-msa.microscopy.com} ; Sun, 15 Apr 2007 22:25:08 -0500 9, 26 -- Received: from yourb27fb1c401 (c-71-230-36-118.hsd1.pa.comcast.net [71.230.36.118]) 9, 26 -- (authenticated bits=0) 9, 26 -- by diskless5.axs2000.net (8.12.11/8.12.11) with ESMTP id l3G3P7UO001573 9, 26 -- for {microscopy-at-msa.microscopy.com} ; Sun, 15 Apr 2007 23:25:08 -0400 9, 26 -- X-IDV-FirstRcvd: c-71-230-36-118.hsd1.pa.comcast.net [71.230.36.118] 9, 26 -- X-IDV-HELO: yourb27fb1c401 9, 26 -- X-IDV-Authenticated-User: cgarber 9, 26 -- Message-ID: {077701c77fd6$d9bc8c90$6501a8c0-at-yourb27fb1c401} 9, 26 -- From: "Garber, Charles A" {cgarber-at-2spi.com} 9, 26 -- To: "Microscopy Listserver" {microscopy-at-msa.microscopy.com} 9, 26 -- Subject: Special microscope slide request 9, 26 -- Date: Sun, 15 Apr 2007 23:25:15 -0400 9, 26 -- MIME-Version: 1.0 9, 26 -- Content-Type: text/plain; 9, 26 -- format=flowed; 9, 26 -- charset="iso-8859-1"; 9, 26 -- reply-type=original 9, 26 -- Content-Transfer-Encoding: 7bit 9, 26 -- X-Priority: 3 9, 26 -- X-MSMail-Priority: Normal 9, 26 -- X-Mailer: Microsoft Outlook Express 6.00.2900.3028 9, 26 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.3028 ==============================End of - Headers==============================
You can get depleted (or at least you used to be able to get this) UAc so that your safety people (or whoever) can be assured that you are not getting into making nuclear warheads, etc. Of course, I suppose that trying to reason with them by noting the very small amount you would be receiving could not be used for such purposes, plus it is not enriched U but natural isotopes.... Probably too much to hope that reason and logic has any place in such discussions.
Roger Moretz, Ph.D. Dept. of Toxicology Boehringer Ingelheim Pharmaceuticals, Inc.
Disclaimer: The comments and opinions are those of the author alone, and do not reflect any official position by his employer.
On 4/13/07, cervantes-at-bendres.com {cervantes-at-bendres.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 } ---------------------------------------------------------------------------- } } Due to some bureaucratic mumbo-jumbo, I am not able to use Uranyl } Acetate (nuclear warheads, blah, blah, blah). I would like to stain } some osmium-fixed tissue thin-sections for TEM; the procedure I'm } following has a UA/Sato's lead stain step for the sections. Does anyone } know of a suitable alternative? I did a quick google and listserver } archive search and didn't find anything, but I'm hoping someone has run } into this problem before and can suggest something. } } Crossing my fingers, } Jessica Cervantes } Bend Research, Inc } Bend, OR } } } ==============================Original Headers============================== } 3, 16 -- From cervantes-at-bendres.com Fri Apr 13 15:12:47 2007 } 3, 16 -- Received: from BRIEX04A.bri.local (bc161112.bendcable.com [216.228.161.112]) } 3, 16 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3DKClG6024466 } 3, 16 -- for {Microscopy-at-microscopy.com} ; Fri, 13 Apr 2007 15:12:47 -0500 } 3, 16 -- MIME-Version: 1.0 } 3, 16 -- Content-Type: text/plain; } 3, 16 -- charset="us-ascii" } 3, 16 -- Subject: TEM: Alternative to Uranyl Acetate Stain } 3, 16 -- Date: Fri, 13 Apr 2007 13:12:46 -0700 } 3, 16 -- Message-ID: {8943D65F9AD70E4488AD6DE09F15088768C44D-at-BRIEX04A} } 3, 16 -- In-Reply-To: {8943D65F9AD70E4488AD6DE09F15088768C44C-at-BRIEX04A} } 3, 16 -- References: {8943D65F9AD70E4488AD6DE09F15088768C44C-at-BRIEX04A} } 3, 16 -- From: "Cervantes, Jessica" {cervantes-at-bendres.com} } 3, 16 -- To: {Microscopy-at-microscopy.com} } 3, 16 -- Content-Transfer-Encoding: 8bit } 3, 16 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l3DKClG6024466 } ==============================End of - Headers============================== }
We prepare slides for micro-electrochemistry from standard 1X3 glass slides which are masked with a strip of tape over the region that will hold the sample then sputter coated with gold using 3-5X the exposure used for SEM sample prep. Be sure to wrap the tape all the way around the slide and also along the back to avoid conductive paths along the edges and to avoid conduction paths to the stage. We typically use tape cut to 3mm for our cell width and also mask the entire back and edges of the slide. After sputtering, we clean the slide and use a drop of conductive silver paint ( the usual SEM suppliers) to attach flexible lead wires. The samples are applied and covered with a square cover slip.
Our own applications are all low voltage but if you intend to run at high voltages you need to prepare very carefully for microscopist safety. If you are intending to use high voltages, especially electrophoresis supplies, you need to get competent design and audit help from electrical engineers with high voltage safety experience.
James Carnahan
Edison Analytical Laboratories, Inc. 301 Nott Street Schenectady, NY 12305
(518) 393-2112
==============================Original Headers============================== 14, 22 -- From carnahan-at-edison-labs.com Mon Apr 16 07:58:52 2007 14, 22 -- Received: from mail3.dotsterhost.com (mail3.dotsterhost.com [72.5.54.189]) 14, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id l3GCwpA6014605 14, 22 -- for {microscopy-at-microscopy.com} ; Mon, 16 Apr 2007 07:58:51 -0500 14, 22 -- Received: (qmail 5698 invoked from network); 16 Apr 2007 12:58:46 -0000 14, 22 -- Received: from unknown (HELO Edisonlab) (carnahan-at-edison-labs.com-at-[141.149.7.9]) 14, 22 -- by 72.5.54.189 with SMTP; 16 Apr 2007 12:58:46 -0000 14, 22 -- Message-ID: {000b01c78026$cda08470$2e01a8c0-at-Edisonlab} 14, 22 -- From: "Jim Carnahan" {carnahan-at-edison-labs.com} 14, 22 -- To: {microscopy-at-microscopy.com} 14, 22 -- Subject: RE: Potential difference across microscope slide. 14, 22 -- Date: Mon, 16 Apr 2007 07:57:33 -0500 14, 22 -- MIME-Version: 1.0 14, 22 -- Content-Type: text/plain; 14, 22 -- format=flowed; 14, 22 -- charset="iso-8859-1"; 14, 22 -- reply-type=original 14, 22 -- Content-Transfer-Encoding: 7bit 14, 22 -- X-Priority: 3 14, 22 -- X-MSMail-Priority: Normal 14, 22 -- X-Mailer: Microsoft Outlook Express 6.00.2900.2869 14, 22 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2869 ==============================End of - Headers==============================
Thanks to everyone who responded. I have indeed brought up the fact that the UA is from depleted uranium, and will continue to try to reason with people. I have managed to get sodium cacodylate (it contains arsenic!) and OsO4 (can blacken your eyeball!) in the door, so there is some hope . . . in the meantime at least now I have some alternatives to try.
Thanks again.
Jessica Cervantes Bend Research, Inc Bend, OR
==============================Original Headers============================== 5, 14 -- From cervantes-at-bendres.com Mon Apr 16 10:57:40 2007 5, 14 -- Received: from BRIEX04A.bri.local (bc161112.bendcable.com [216.228.161.112]) 5, 14 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3GFvc4L030205 5, 14 -- for {Microscopy-at-microscopy.com} ; Mon, 16 Apr 2007 10:57:38 -0500 5, 14 -- MIME-Version: 1.0 5, 14 -- Content-Type: text/plain; 5, 14 -- charset="us-ascii" 5, 14 -- Subject: TEM:Alternative to Uranyl Acetate Stain 5, 14 -- Date: Mon, 16 Apr 2007 08:57:36 -0700 5, 14 -- Message-ID: {8943D65F9AD70E4488AD6DE09F15088768C458-at-BRIEX04A} 5, 14 -- From: "Cervantes, Jessica" {cervantes-at-bendres.com} 5, 14 -- To: {Microscopy-at-microscopy.com} 5, 14 -- Content-Transfer-Encoding: 8bit 5, 14 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l3GFvc4L030205 ==============================End of - Headers==============================
I'm processing some monolayer cell cultures grown on Thermanox coverslips for SEM. I seem to recall that aldehyde fixatives can cause an artifact that results in small holes in the cell membrane. If I remember, Picric Acid is added to the fixative and that it helps prevent the artifact. I don't remember where I originally read this. Am I off base on this? If it is correct, does anyone out there remember the formulation of the fixative using Picric Acid? I currently use 2% glutaraldehyde, 2% paraformaldehyde and 0.5% acrolein in 0.1M Sorensen's phosphate buffer 7.2pH.
Also during processing the thin cytoplasmic extensions of the cells break. I assume this is due to shrinkage during the dehydration steps and critical point drying. Anyone know of a way to prevent this?
All help is appreciated. Thanks.
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============================== 7, 20 -- From tbargar-at-unmc.edu Mon Apr 16 10:58:42 2007 7, 20 -- Received: from zixvpm01.unmc.edu (zixvpm01.unmc.edu [192.198.54.126]) 7, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3GFwgQi032217 7, 20 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 16 Apr 2007 10:58:42 -0500 7, 20 -- Received: from zixvpm01.unmc.edu (ZixVPM [127.0.0.1]) 7, 20 -- by Outbound.unmc.edu (Proprietary) with ESMTP id 278A54C0CE 7, 20 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 16 Apr 2007 10:58:42 -0500 (CDT) 7, 20 -- Received: from unmcnotes01.unmc.edu (host-137-197-103-35.unmc.edu [137.197.103.35]) 7, 20 -- by zixvpm01.unmc.edu (Proprietary) with ESMTP id 102DA4C0CB 7, 20 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 16 Apr 2007 10:58:41 -0500 (CDT) 7, 20 -- Subject: cell cultures and aldehyde fixatives 7, 20 -- To: Microscopy-at-MSA.Microscopy.Com 7, 20 -- X-Mailer: Lotus Notes Release 7.0.1 January 17, 2006 7, 20 -- Message-ID: {OF082F625E.F717EFD3-ON862572BF.00568FD6-862572BF.0057C42F-at-unmc.edu} 7, 20 -- From: Tom W Bargar {tbargar-at-unmc.edu} 7, 20 -- Date: Mon, 16 Apr 2007 10:58:38 -0500 7, 20 -- X-MIMETrack: Serialize by Router on UNMCNOTES01.UNMC.EDU/Servers/UNEBR at 04/16/2007 10:58:41 7, 20 -- AM 7, 20 -- MIME-Version: 1.0 7, 20 -- Content-type: text/plain; charset=US-ASCII ==============================End of - Headers==============================
For the membrane holes, try 1% tannic acid in the glut. You want the monomeric form, Mallinckrodt 1674 - or 1764, I keep transposing those digits. 1% T.A. can also be used in an OsO4 post-fix. I would also suggest cutting the glut to 1 or 1.25%, and don't bother with the formalin, it's not neeed for cell monolayers. Acrolein I haven't used, so can't comment. I've just done 1-1.25% glut + 1% tannic acid. I don't know about picric acid, and would be interested to hear if it works.
Phil
} Dear listers, } } I'm processing some monolayer cell cultures grown on Thermanox coverslips } for SEM. I seem to recall that aldehyde fixatives can cause an artifact } that results in small holes in the cell membrane. If I remember, Picric } Acid is added to the fixative and that it helps prevent the artifact. I } don't remember where I originally read this. Am I off base on this? If it } is correct, does anyone out there remember the formulation of the fixative } using Picric Acid? I currently use 2% glutaraldehyde, 2% paraformaldehyde } and 0.5% acrolein in 0.1M Sorensen's phosphate buffer 7.2pH. } } Also during processing the thin cytoplasmic extensions of the cells break. } I assume this is due to shrinkage during the dehydration steps and critical } point drying. Anyone know of a way to prevent this? } } All help is appreciated. Thanks. } } 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 -- 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============================== 4, 22 -- From oshel1pe-at-cmich.edu Mon Apr 16 11:44:16 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 l3GGiGuD001107 4, 22 -- for {Microscopy-at-microscopy.com} ; Mon, 16 Apr 2007 11:44:16 -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 l3GH8e6j012202 4, 22 -- for {Microscopy-at-microscopy.com} ; Mon, 16 Apr 2007 13:08:41 -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 -- Mon, 16 Apr 2007 12:44:14 -0400 4, 22 -- Mime-Version: 1.0 4, 22 -- Message-Id: {f06230909c2495730d7d2-at-[141.209.160.249]} 4, 22 -- In-Reply-To: {200704161604.l3GG4XD5018655-at-ns.microscopy.com} 4, 22 -- References: {200704161604.l3GG4XD5018655-at-ns.microscopy.com} 4, 22 -- Date: Mon, 16 Apr 2007 12:44:11 -0400 4, 22 -- To: Microscopy-at-microscopy.com 4, 22 -- From: Philip Oshel {oshel1pe-at-cmich.edu} 4, 22 -- Subject: Re: [Microscopy] cell cultures and aldehyde fixatives 4, 22 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 4, 22 -- X-OriginalArrivalTime: 16 Apr 2007 16:44:14.0179 (UTC) FILETIME=[7759D330:01C78046] 4, 22 -- X-CanItPRO-Stream: default 4, 22 -- X-Spam-Score: -4 () L_EXCH_MF 4, 22 -- X-Scanned-By: CanIt (www . roaringpenguin . com) on 141.209.20.21 ==============================End of - Headers==============================
HI Tom- I use picric acid in my primary fix to preserve membranes. My "recipe" is 2.5% glut, 4% pfa in 0.1M Na-cacod. with 0.02% picric acid. I did the math years ago, and this worked out to be: 1 vial of 10% glut + 1 vail of 16% pfa in 20 ml of 0.2m cacod with 2 ml of saturated aqueous picric acid added. I've had the same 100g bottle of picric acid for over 15 years. I just keep it saturated with the liquid well above the level of the crystals. When I draw some off, I add more water. Our Life Safety guys here are just thrilled with me: osmium, uranium picric acid, suspected carcinogens ...all the fun stuff we EM folk play with. As long as you are careful about keeping the crystals fully under water, and not letting any accumulate around the rim or anywhere where they could dry out, you're fine.
As for your broken processes: they may be getting beaten up during your CPD run. Be sure to do your CO2 exchanges gently, never letting the fluid level drop below your sample (this will mean extra exchanges), and then, at the end, vent very slowly...100 psi/minute or less.
Good luck! Lee -- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy & Histology Core Facility Manager, Optical Microscopy Core Facility Weill-Cornell Medical College
We are trying to stabilize lysosomes for section surface immunocytochemisty and are searching for a fixation method which might best preserve the membranes without destroying antigenicity. We are working with cultured cells embedded in LR White. Our usual method of 4% paraformaldehyde with 1.0% glut (buffered with culture media) appears to allow stabilization thru approximately 70% EtOH, but the lysosomes appear to break afterwards, spilling their contents into the cytoplasm. Fixation and dehydration were done on ice. We are considering a PLT procedure, perhaps also including a little Uranyl acetate in the fixative. Are there any suggestions for preserving these delicate membranes?
Many thanks,
Doug
Douglas R. Keene Assistant Investigator Micro-Imaging Center Shriners Hospital for Children 3102 S.W. Sam Jackson Park Road Portland, Oregon 97239 503-221-3434 drk-at-shcc.org
==============================Original Headers============================== 7, 21 -- From drk-at-SHCC.org Mon Apr 16 12:05:51 2007 7, 21 -- Received: from mail.SHCC.org (mail.shcc.org [64.213.211.202]) 7, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3GH5pf5024494 7, 21 -- for {Microscopy-at-Microscopy.Com} ; Mon, 16 Apr 2007 12:05:51 -0500 7, 21 -- Received: from DRK2 ([64.213.211.62]) by mail.SHCC.org (PMDF V6.3 #31473) 7, 21 -- with ESMTPA id {0JGL0089AORQ16-at-mail.SHCC.org} for Microscopy-at-Microscopy.Com; 7, 21 -- Mon, 16 Apr 2007 10:04:38 -0700 (PDT) 7, 21 -- Date: Mon, 16 Apr 2007 10:06:24 -0700 7, 21 -- From: Doug Keene {drk-at-SHCC.org} 7, 21 -- Subject: fixation of lysosome membranes 7, 21 -- Sender: drk-at-SHCC.org 7, 21 -- To: Microscopy-at-Microscopy.Com 7, 21 -- Reply-to: drk-at-SHCC.org 7, 21 -- Message-id: {0JGL0089BORQ16-at-mail.SHCC.org} 7, 21 -- Organization: Shriners Hospitals for Children 7, 21 -- MIME-version: 1.0 7, 21 -- X-MIMEOLE: Produced By Microsoft MimeOLE V6.00.2900.3028 7, 21 -- X-Mailer: Microsoft Office Outlook, Build 11.0.5510 7, 21 -- Content-type: text/plain; charset=us-ascii 7, 21 -- Content-transfer-encoding: 7BIT 7, 21 -- Thread-Index: AceASZBVyiHQEEImTY6DUEfJOjWm8A== ==============================End of - Headers==============================
A couple of you have asked what the responses were to my query for alternatives to UA. Here's the list:
1. Bismuth Staining - from Mike Nesson
Quote: There are several references that deal with Bismuth staining. I tried some of these years ago and was quite satisfied with the results.
Locke, M and Huie, P. "Bismuth staining for light and electron microscopy" Tissue and Cell: 9; 347-371 (1977).
2. Straight lead citrate - from Ted, Managing Director, The EMscope Company Ltd., Thailand.
3. Nanovan (a negative stain)- from David Gene Morgan, University of California at Davis
Quote: There is a product called nanovan (sold by Nanoprobes) which is a methylamine vanadate stain that has similar features to uranly acetate. The manufacturers claim finer grain size and some other benefits, if I remember correctly. The only uses I am aware of have been to replace simple uranyl acetate staining of isolated biological materials, and I have no idea if it would even work as a TEM post-stain.
4. Reduced Osmium - from Rachid
Quote: There is no need for UA if your sample is treated with reduced osmium (osmium + potassium ferrocyanide). You will just have to contrast 2 min with lead citrate ... the contrast you will get will be really good. You can check on this web site that I am putting together ... all pictures in the gallery are produced as I mentioned. http://rsougrat.googlepages.com/
5. KMnO4 and/or Tannic Acid - from Mike Reedy
Quote: You will get even more contrast if you use KMnO4 followed by Sato's, as we have since 1964 (when Pb was was Pb cittrate, not Sato's). And, you can get good contrast on thinner sections. Using Tannic acid in the block stain, before OSO4, or after but followed by UrAc, adds to contrast, improves preservation. We have had excellent results (see attached).
Some reports of using TA as a section stain before Pb stain can also be found with google help. I've never tried it yet.
KMnO4 section stain won't work if NMA is in your Epon mix. End quote.
Mike included attachments in his reply, which can't be posted.
6. Tell the safety people to get a brain - from several people
Thanks again to everybody who responded. This is a great resource.
Jessica Cervantes Bend Research Inc Bend, OR
==============================Original Headers============================== 25, 14 -- From cervantes-at-bendres.com Mon Apr 16 12:57:04 2007 25, 14 -- Received: from BRIEX04A.bri.local (bc161112.bendcable.com [216.228.161.112]) 25, 14 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3GHv4AU004437 25, 14 -- for {Microscopy-at-microscopy.com} ; Mon, 16 Apr 2007 12:57:04 -0500 25, 14 -- MIME-Version: 1.0 25, 14 -- Content-Type: text/plain; 25, 14 -- charset="us-ascii" 25, 14 -- Subject: TEM:Alternatives to Uranyl Acetate Stain - Responses to the Original Query 25, 14 -- Date: Mon, 16 Apr 2007 10:57:03 -0700 25, 14 -- Message-ID: {8943D65F9AD70E4488AD6DE09F15088768C45E-at-BRIEX04A} 25, 14 -- From: "Cervantes, Jessica" {cervantes-at-bendres.com} 25, 14 -- To: {Microscopy-at-microscopy.com} 25, 14 -- Content-Transfer-Encoding: 8bit 25, 14 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l3GHv4AU004437 ==============================End of - Headers==============================
In 1964, I published a small note about the potential use of Indium Trichloride as a stain during embedding (J de Microscopie, 3: pp575- 578). As I recall, it added some contrast, specifically to virus structures. It might be worth considering.
Joel
Date sent: Mon, 16 Apr 2007 12:57:11 -0500 To: jbs-at-temple.edu X-from: cervantes-at-bendres.com Send reply to: cervantes-at-bendres.com
Hi All, Jan Leunissen pointed out that I neglected to include the original volumes of the vials of pfa and glut...sorry about that. I buy 10ml vials of both. Lee -- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy & Histology Core Facility Manager, Optical Microscopy Core Facility Weill-Cornell Medical College
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 rpowell-at-nanoprobes.com as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: rpowell-at-nanoprobes.com Name: Rick Powell
Organization: Nanoprobes, Incorporated
Title-Subject: [Microscopy] TEM:Alternatives to Uranyl Acetate Stain
Question: Response from commercial vendor (Nanoprobes):
Hello Everyone:
We do offer "NanoVan" as a commercial product. It is based on methylamine vanadate, which has a lower atomic number than uranium, and will give a ligher stain (since we make small gold particles, this is helpful).
We also offer Nano-W, an alternative based on tungsten which produces a denser stain. The two may be combined for intermiediate stain densities.
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Email: DLowry-at-asu.edu Name: David Lowry
Organization: Arizona State University
Title-Subject: [Filtered] Mycobacterium prep for immuno-label
Question: I am attempting to do thin section immuno-labeling experiments with Mycobacterium tuberculosis. I have used standard procedures: fixation with 4% paraformaldehyde/0.1% glut, then enmeshed cells in ~ 1% agarose prior to EtOH dehydration and LR White resin. Very basic stuff.
Unfortunately, in 2 separate trials I have encountered a problem with very large holes in the resin around and between the clusters of cells. It seems to be either incomplete dehydration or poor penetration of resin, although in both attempts I used 100% EtOH dehydration and extensive time in 100% LR.
Are there special considerations to take when working with this organism? I have searched on-line for information but could not locate any specific indications. I suspect there may be something unusual about the outer cell boundary that is causing this problem, and would like to know if anyone may have experience in dealing with this organism.
Thanks Vlad. I did realize that some of these alternatives would not work for my application, but wanted to list them for others (ie, Nanovan is an alternative to UA as a negative stain, not for sections, as you point out). I do plan on continuing to educate people here on UA, but sometimes these things are political, rather than scientific.
Thanks again, Jessica Cervantes
-----Original Message----- X-from: Vlad Speransky [mailto:vladislav_speransky-at-nih.gov] Sent: Monday, April 16, 2007 11:59 AM To: Cervantes, Jessica
Dear colleagues,
Usually to observe apopototic patterns in fluorescence microscopy the most straigthforward method is to label the cell nuclei with Hoechst. I very rarely noticed that DAPI was used. Why? Is there a reason why Hoechst should be preferred to DAPI?
Stephane, epifluorescer not confocaler (or is it confocaliser?)
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Anyone have a copy in good or better condition of Hayat & Miller, "Negative Staining" that they want to sell? I can only find one on the web at an outrageous price. This is for the lab out of my pocket, so price is also important. Be nice if they put out a 2nd edition (hint, hint). Thanks.
Phil -- 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
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The next meeting of the Midwest Microscopy and Microanalysis Society, an Optical Techniques Workshop, will be held on Thursday, May 17, at the College of Microscopy in Westmont, IL (The McCrone Group). Please follow the link below and click on Meetings for program details and registration information.
www.midwestmicroscopy.org
We look forward to seeing you there.
Elaine Schumacher President, Midwest Microscopy and Microanalysis Society elaine-at-midwestmicroscopy.org
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Email: rpowell-at-nanoprobes.com Name: Rick Powell
Organization: Nanoprobes, Incorporated
Title-Subject: [Filtered] TEM:Alternatives to Uranyl Acetate Stain
Question: Hello Everyone:
In my somewhat hasty response to Jessica Cervantes, I had overlooked the fact that the original post referred to poststaining (positive staining) of sections. To clarify, our NanoVan and Nano-W products are intended as negative stains for protein complexes, viruses, etc.; we are not aware of any references that describe their use on thin sections.
If anyone has tried this, or has seen any references, we would of course very much like to know.
The MSA video catalog is currently unavailable online . It had been hosted at the University of Florida, Biotech Center. They are totally revamping their web site and many things have not yet been restored to active status. I have arranged with Nestor to have the catalog hosted on the MSA server. AS soon as I clean up the file a bit, Nestor will be able to get it up and running and accessible from the MSA main page. In the meantime, if you need any info about the video collection, please feel free to email me. The "Tips & Tricks" page that was hosted by the EM Core Lab at Florida is also offline. I am told that it will eventually be restored. Stay tuned.
Greg -- Greg Erdos University of Florida, Retired Micanopy, Florida
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I have a request from a client who wants to localize a gene locus within the nucleus using FISH but then wants to determine better resolution with TEM. My first reaction would be that the FISH protocol would destroy the ultrastructure but thought I would ask the bigger brain if anyone out there has done something similar. I have read a couple of papers that have done this but I was not impressed with the ultrastructure. All thoughts are appreciated.
Garnet
-- 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============================== 6, 27 -- From gmartens-at-interchange.ubc.ca Tue Apr 17 11:22:48 2007 6, 27 -- Received: from mr2.mail-relay.ubc.ca (mr2.mail-relay.ubc.ca [137.82.45.3]) 6, 27 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3HGMm6w030129 6, 27 -- for {microscopy-at-microscopy.com} ; Tue, 17 Apr 2007 11:22:48 -0500 6, 27 -- Received: from mr2.mail-relay.ubc.ca (localhost [127.0.0.1]) 6, 27 -- by localhost (Postfix) with SMTP id 83D24C3CC 6, 27 -- for {microscopy-at-microscopy.com} ; Tue, 17 Apr 2007 09:22:47 -0700 (PDT) 6, 27 -- Received: from mta2.interchange.ubc.ca (mta2.interchange.ubc.ca [142.103.145.70]) 6, 27 -- by mr2.mail-relay.ubc.ca (Postfix) with ESMTP 6, 27 -- for {microscopy-at-microscopy.com} ; Tue, 17 Apr 2007 09:22:45 -0700 (PDT) 6, 27 -- Received: from [137.82.85.216] (0511.emlab.ubc.ca [137.82.85.216]) 6, 27 -- by smtp.interchange.ubc.ca 6, 27 -- (iPlanet Messaging Server 5.2 HotFix 1.21 (built Sep 8 2003)) 6, 27 -- with ESMTPA id {0JGN00LDPHHWO7-at-smtp.interchange.ubc.ca} for 6, 27 -- microscopy-at-microscopy.com; Tue, 17 Apr 2007 09:22:45 -0700 (PDT) 6, 27 -- Date: Tue, 17 Apr 2007 09:22:43 -0700 6, 27 -- From: Garnet Martens {gmartens-at-interchange.ubc.ca} 6, 27 -- Subject: FISH followed by EM 6, 27 -- To: microscopy-at-microscopy.com 6, 27 -- Message-id: {a06240804c24aa36eb663-at-[137.82.85.216]} 6, 27 -- MIME-version: 1.0 6, 27 -- Content-type: text/plain; format=flowed; charset=us-ascii 6, 27 -- X-UBC-Scanned: Sophos PureMessage 5.3.1.294258, Antispam-Engine: 2.5.0.283055, Antispam-Data: 2007.4.17.91234 6, 27 -- X-UBC-Relayed: Relayed through mail-relay.ubc.ca 6, 27 -- 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 6, 27 -- X-Spam-Level: 6, 27 -- X-Spam-Flag: No ==============================End of - Headers==============================
There is a intriguing new technique that combines AFM with ultramicrotomy that images from the block face. Some of the early work, done by Dr. Anton Efimov of NT-MDT (efimov-at-ntmdt.ru) shows very delicate ultrastructure (~2-6nm structures) . The advantage is that this system uses local differences in elasticity to image, rather than heavy metal staining. You can do serial sections and, while the AFM images from the block face, the slices are available for conventional imaging with any sort of microscopy.
Might be an interesting alternative. Suggest that you write directly to Dr. Efimov for further information.
Best regards, 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 12:05 PM 4/17/2007, gmartens-at-interchange.ubc.ca wrote:
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==============================Original Headers============================== 14, 17 -- From bfoster-at-mme1.com Tue Apr 17 12:11:16 2007 14, 17 -- Received: from 5starpro.com (enterprise.5starpro.com [207.44.136.95]) 14, 17 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3HHBFcA010680 14, 17 -- for {microscopy-at-microscopy.com} ; Tue, 17 Apr 2007 12:11:16 -0500 14, 17 -- Message-Id: {200704171711.l3HHBFcA010680-at-ns.microscopy.com} 14, 17 -- Received: (qmail 8374 invoked by uid 2020); 17 Apr 2007 12:45:09 -0500 14, 17 -- Received: from host57-99.rancor.birch.net (HELO barbsd505.mme1.com) (65.17.57.99) 14, 17 -- by enterprise.5starpro.com with (DHE-RSA-AES256-SHA encrypted) SMTP; 17 Apr 2007 12:45:09 -0500 14, 17 -- X-Mailer: QUALCOMM Windows Eudora Version 7.1.0.9 14, 17 -- Date: Tue, 17 Apr 2007 12:10:51 -0500 14, 17 -- To: gmartens-at-interchange.ubc.ca, microscopy-at-microscopy.com 14, 17 -- From: Barbara Foster {bfoster-at-mme1.com} 14, 17 -- Subject: Re: [Microscopy] FISH followed by EM 14, 17 -- In-Reply-To: {200704171626.l3HGQq1j001551-at-ns.microscopy.com} 14, 17 -- References: {200704171626.l3HGQq1j001551-at-ns.microscopy.com} 14, 17 -- Mime-Version: 1.0 14, 17 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
Jessica Don't know about KMnO4 staining compatibility with LR White; please let me know. The easy test is to put a blank block or chunk or flake of cured LR White in 1-2% KMnO4 and soak it for an hour or a day to see if resin surface turns light brown, dark brown, or black and charred looking. No reaction is good news for section staining.
The more stringent assay for reaction is to put the test tube of KMnO4 solution with the cured embedding resin in a beaker of water and then heat the water to boiling for an hour or so, cool it off and examine the resin surface for such changes. Araldite 506 (lowest viscosity one I know of) and other Araldites used for embedding are remarkable resistant to surface discoloration when this is done. Epon DDSA (no MNA; fiddle the mix until it is hard enough to please you, and forget the epoxy:anhydride ratio lore) turns moderately browner, as I recall, but is still acceptable for section staining, with some tendency to be more granular and maybe show some hard-to-eliminate nano-pepper stain deposit in contrast to Araldite.
LR White is an acrylic says EMS: LR White is a polar monomer polyhydroxylated acromatic acrylic resin. It can be cured by heat or by UV light. Sections of polymerized LR White resin are hydrophilic I think Lawn's 1960 JCB paper introducing KMnO4 section staining used sections of methacrylate, the mix of methyl and butyl methacrylate we all used in the late 1950s before epoxies, especially Epon, turned up and became dominant.
BTW-- tannic acid is a fixative and a mordant, and magically capable of superior structure preservation and assuring really strong uniform staining in our hands-- see the evidence of 13Å preservtion in fiber x-ray diffraction from fixed-embedded fibers in Sader et al 2007 in J Struct Biol (Articles In Press on line), and look at EMs in some reprints I sent you for evidence of the good morphology. The latter is a wondrous gift of TA I had no idea of until I learned it from David Begg et al, J. Cell Biol. 79:846-852, an all-time key paper in my book of methodological turning points.. The other key was the observation by Hirose and Wakabayashi (J. Mol. Biol. 204:797-801) that TA followed by OsO4 or UrAc give great morphological fixation without aldehydes. They used it for freeze-substitution, as we have, but we found it also worked very well as a general fix (we termed this TAURAC) for permeabilized cells in aqueous buffers so long as we exluded TA blockers like PVP, Triton X 100 etc.
Your UA police might be interested in a demo of how completely tannins, esp tannic acid, can convert a UrAc solution into a flocculent brown precipitate at the bottom of a UrAc-free supernatan (it LOOKS UrAc free! I made no measurements.). TA is cheap in non-EM grades; maybe they'd accept precipitation as a way of rendering it safely bio-inactive. From the information online at EMS ib their datasheet/22400, I've estimated that the 10,400 counts/secin depleted uranium is reduced to about -2 ct/sec in a very small bundle of muscle fibers containing 1 ug of protein and binding maybe 5 ug of UrAc. But it still requires health police and special containment if such a specimen is to be transported into or within the DOE facility at Argonne National Laboratory. The inconvenience has so far discouraged me from pursuing some experiments that would require such a specimen, but one day I will do it, legally and according to their regulations. I doubt a Geiger counter could detect any rise above background in the presence of that small an amount of UrAc.
-mike-
} Thanks for this Mike. I think KMnO4 followed by Sato's is one of the } better alternatives I've seen so far. Your results are very impressive. } } } Any idea if KMnO4 will work with LR White embedded tissues? } } Thanks, } Jessica Cervantes } Bend Research Inc } } -----Original Message----- } From: Mike Reedy [mailto:mike.reedy-at-cellbio.duke.edu] } Sent: Saturday, April 14, 2007 5:19 PM } To: Cervantes, Jessica } Subject: Re: [Microscopy] TEM: Alternative to Uranyl Acetate Stain } } You will get even more contrast if you use KMnO4 followed by Sato's, } as we have since 1964 (when Pb was was Pb cittrate, not Sato's). } And, you can get good contrast on thinner sections. Using Tannic } acid in the block stain, before OSO4, or after but followed by UrAc, } adds to contrast, improves preservation. We have had excellent } results (see attached). } } Some reports of using TA as a section stain before Pb stain can also } be found with google help. I've never tried it yet. } } KMnO4 section stain won't work if NMA is in your Epon mix. see } attached. } -mike reedy- } } ----------------------------------------------------------------------- } ----- } } 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
Jessica sorry to resend all that. I am trying to see if I can calm the HTML detector to break through into the list server, -- -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
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Question: Please join us for the New England Society of Microscopy's (NESM)24th Annual Woods Hole Meeting, where we will be celebrating NESM's 40th birthday!
Expect very exciting talks and a special presentation from most of the past presidents.
Please see the link to the newsletter for more information:
Been away so missed the original question but has any one suggested p- Phenylenediamine?
Numerous references to using p-phenylenediamine but one to start with is "The use of p-phenylenediamine in the block to enhance osmium staining for electron microscopy" Stain Technology, Vol 47, No5 pp 239 - 243.
Added in the 70% ethanol dehydration step from memory.
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
EM Centre: http://ocem.otago.ac.nz/ Confocal Centre: http://occm.otago.ac.nz/ Department: http://anatomy.otago.ac.nz/
==============================Original Headers============================== 12, 20 -- From allan.mitchell-at-stonebow.otago.ac.nz Tue Apr 17 22:11:32 2007 12, 20 -- Received: from mailhub2.otago.ac.nz (mailhub2.otago.ac.nz [139.80.64.247]) 12, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3I3BUnE007364 12, 20 -- for {microscopy-at-msa.microscopy.com} ; Tue, 17 Apr 2007 22:11:31 -0500 12, 20 -- Received: from galadriel.otago.ac.nz (galadriel.otago.ac.nz [139.80.64.213]) 12, 20 -- by mailhub2.otago.ac.nz (8.13.8/8.13.8) with ESMTP id l3I3BTuh021138 12, 20 -- for {microscopy-at-msa.microscopy.com} ; Wed, 18 Apr 2007 15:11:29 +1200 12, 20 -- Received: from allan.otago.ac.nz ([139.80.40.92]) 12, 20 -- by galadriel.otago.ac.nz with esmtp (Exim 4.50) 12, 20 -- id 1He0Zp-00036T-6z 12, 20 -- for microscopy-at-msa.microscopy.com; Wed, 18 Apr 2007 15:11:25 +1200 12, 20 -- Mime-Version: 1.0 (Apple Message framework v752.2) 12, 20 -- Content-Transfer-Encoding: 7bit 12, 20 -- Message-Id: {F4CB3CDE-DFAC-42EA-9A9E-1B924398D2C3-at-stonebow.otago.ac.nz} 12, 20 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 12, 20 -- To: microscopy-at-msa.microscopy.com 12, 20 -- From: Allan Mitchell {allan.mitchell-at-stonebow.otago.ac.nz} 12, 20 -- Subject: [Microscopy] RE: TEM: Alternative to Uranyl Acetate Stain 12, 20 -- Date: Wed, 18 Apr 2007 15:11:27 +1200 12, 20 -- X-Mailer: Apple Mail (2.752.2) ==============================End of - Headers==============================
I want to detect aluminosilicate particles in the middle of organic material. The particles are expected to be too small for light microscopy and too dilute for TEM. The solution would be to dry everything flat on a SEM stub and to find a way to differentiate organic particles for aluminosilicate particles. Our EDX doesn't want to start so I wondered if I could see something with BSE? X-from the atomic weight of the elements, Al and Si are not particularly heavy but they are of course very dense in the particles. Do you think it would be possible? Any remark?
Stephane
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if you own a decent BSE detector, I'm sure that you will be able to see a contrast between the two types of particles. Average atomic number is well apart from each other. To make the observation easier, I would recommend to use a low-Z SEM stub like graphite or a graphite plate on top of a standard holder. Then you will be able to see your silicates with a bright contrast with respect to the background and the other particles.
Best regards,
Petra --------------------------------------- Dr. Petra Wahlbring Goodyear S.A. Technical Center Analytical Test Laboratories L-7750 Colmar-Berg Luxembourg Tel +352 8199 3725 Fax +352 8199 3905 e-mail: petra.wahlbring-at-goodyear.com
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04/18/07 03:09 PM To petra.wahlbring-at-goodyear.com cc Please respond to nizets2-at-yahoo.com Subject [Microscopy] aluminosilicate and BSE
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Dear Listers,
I want to detect aluminosilicate particles in the middle of organic material. The particles are expected to be too small for light microscopy and too dilute for TEM. The solution would be to dry everything flat on a SEM stub and to find a way to differentiate organic particles for aluminosilicate particles. Our EDX doesn't want to start so I wondered if I could see something with BSE? X-from the atomic weight of the elements, Al and Si are not particularly heavy but they are of course very dense in the particles. Do you think it would be possible? Any remark?
Stephane
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Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (tauria-at-hotmail.com) from http://www.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Wednesday, April 11, 2007 at 12:33:00 ---------------------------------------------------------------------------
Email: tauria-at-hotmail.com Name: DR. FRANCIS J. PRONESTI
Organization: WORLD ENERGY SERVICES, LTD.
Education: Graduate College
Location: castellana grotte, bari, italy
Question: HELLO EVERYONE, I NEED URGENTLY AN OPERATING MANUAL FOR A PERKIN ELMER INFRARED MICROSCOPE,MODEL FT-IR, S/N 144235 MANUFACTURED IN 1991. PART NO. N187-3065. THANKS AND KIND REGARDS TO ALL. DR. FRANCIS J. PRONESTI
This Question was submitted to Ask-A-Microscopist by (akoorts-at-medic.up.ac.za) from http://www.msa.microscopy.org/Ask-A-Microscopist/Ask-A-Microscopist.html on Wednesday, April 18, 2007 at 06:36:15 Remember to consider the Grade/Age of the student when considering the Question --------------------------------------------------------------------------- Please reply to both akoorts-at-medic.up.ac.za as well as to the Microscopy Listserver ---------------------------------------------------------------------------
Allan Micthell suggested para-phenylene diamine as an osmium 'enhancer' and I agree. I have used p-pd in 70% ethanol during dehydration for years, it chemically reduces osmium bound to the tissue and increases contrast. The only drawback is that tissue so treated is difficult to stain with the usual toluidine blue + borax solution.
Geoff
allan.mitchell-at-stonebow.otago.ac.nz 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, 36 -- From mcauliff-at-umdnj.edu Wed Apr 18 08:58:19 2007 8, 36 -- Received: from zix01.umdnj.edu (zix01.UMDNJ.EDU [130.219.34.124]) 8, 36 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3IDwII0027754 8, 36 -- for {microscopy-at-msa.microscopy.com} ; Wed, 18 Apr 2007 08:58:19 -0500 8, 36 -- Received: from zix01.umdnj.edu (ZixVPM [127.0.0.1]) 8, 36 -- by Outbound.umdnj.edu (Proprietary) with ESMTP id E5592A7B9C 8, 36 -- for {microscopy-at-msa.microscopy.com} ; Wed, 18 Apr 2007 09:58:17 -0400 (EDT) 8, 36 -- Received: from umdnj.edu (imail2.UMDNJ.EDU [130.219.34.139]) 8, 36 -- by zix01.umdnj.edu (Proprietary) with ESMTP id BDCC4A7B80 8, 36 -- for {microscopy-at-msa.microscopy.com} ; Wed, 18 Apr 2007 09:58:16 -0400 (EDT) 8, 36 -- Received: from ([130.219.34.133]) 8, 36 -- by imail2.umdnj.edu with ESMTP id KP-BRCRF.95415871; 8, 36 -- Wed, 18 Apr 2007 09:57:44 -0400 8, 36 -- Received: from conversion-daemon.Polaris.umdnj.edu by Polaris.umdnj.edu 8, 36 -- (iPlanet Messaging Server 5.2 HotFix 2.02 (built Oct 21 2004)) 8, 36 -- id {0JGP004015FKRL-at-Polaris.umdnj.edu} (original mail from mcauliff-at-umdnj.edu) 8, 36 -- for microscopy-at-msa.microscopy.com; Wed, 18 Apr 2007 09:57:44 -0400 (EDT) 8, 36 -- Received: from [127.0.0.1] ([10.138.2.123]) 8, 36 -- by Polaris.umdnj.edu (iPlanet Messaging Server 5.2 HotFix 2.02 (built Oct 21 8, 36 -- 2004)) with ESMTP id {0JGP009NP5FJOR-at-Polaris.umdnj.edu} ; Wed, 8, 36 -- 18 Apr 2007 09:57:19 -0400 (EDT) 8, 36 -- Date: Wed, 18 Apr 2007 09:58:55 -0400 8, 36 -- From: Geoff McAuliffe {mcauliff-at-umdnj.edu} 8, 36 -- Subject: Re: [Microscopy] Alternative to Uranyl Acetate Stain/p-phenylene 8, 36 -- diamine 8, 36 -- In-reply-to: {200704180312.l3I3CsmO009606-at-ns.microscopy.com} 8, 36 -- To: allan.mitchell-at-stonebow.otago.ac.nz, 8, 36 -- MicroscopyListserver {microscopy-at-msa.microscopy.com} 8, 36 -- Message-id: {4626241F.3040902-at-umdnj.edu} 8, 36 -- MIME-version: 1.0 8, 36 -- Content-type: text/plain; format=flowed; charset=ISO-8859-1 8, 36 -- Content-transfer-encoding: 7BIT 8, 36 -- X-Accept-Language: en-us, en 8, 36 -- User-Agent: Mozilla/5.0 (Windows; U; Windows NT 5.1; en-US; rv:1.7.2) 8, 36 -- Gecko/20040804 Netscape/7.2 (ax) 8, 36 -- References: {200704180312.l3I3CsmO009606-at-ns.microscopy.com} ==============================End of - Headers==============================
Jessica Don't know about KMnO4 staining compatibility with LR White; please let me know. The easy test is to put a blank block or chunk or flake of cured LR White in 1-2% KMnO4 and soak it for an hour or a day to see if resin surface turns light brown, dark brown, or black and charred looking. No reaction is good news for section staining.
The more stringent assay for reaction is to put the test tube of KMnO4 solution with the cured embedding resin in a beaker of water and then heat the water to boiling for an hour or so, cool it off and examine the resin surface for such changes. Araldite 506 (lowest viscosity one I know of) and other Araldites used for embedding are remarkable resistant to surface discoloration when this is done. Epon DDSA (no MNA; fiddle the mix until it is hard enough to please you, and forget the epoxy:anhydride ratio lore) turns moderately browner, as I recall, but is still acceptable for section staining, with some tendency to be more granular and maybe show some hard-to-eliminate nano-pepper stain deposit in contrast to Araldite.
LR White is an acrylic says EMS: LR White is a polar monomer polyhydroxylated acromatic acrylic resin. It can be cured by heat or by UV light. Sections of polymerized LR White resin are hydrophilic I think Lawn's 1960 JCB paper introducing KMnO4 section staining used sections of methacrylate, the mix of methyl and butyl methacrylate we all used in the late 1950s before epoxies, especially Epon, turned up and became dominant.
BTW-- tannic acid is a fixative and a mordant, and magically capable of superior structure preservation and assuring really strong uniform staining in our hands-- see the evidence of 13Å preservtion in fiber x-ray diffraction from fixed-embedded fibers in Sader et al 2007 in J Struct Biol (Articles In Press on line), and look at EMs in some reprints I sent you for evidence of the good morphology. The latter is a wondrous gift of TA I had no idea of until I learned it from David Begg et al, J. Cell Biol. 79:846-852, an all-time key paper in my book of methodological turning points.. The other key was the observation by Hirose and Wakabayashi (J. Mol. Biol. 204:797-801) that TA followed by OsO4 or UrAc give great morphological fixation without aldehydes. They used it for freeze-substitution, as we have, but we found it also worked very well as a general fix (we termed this TAURAC) for permeabilized cells in aqueous buffers so long as we exluded TA blockers like PVP, Triton X 100 etc.
Your UA police might be interested in a demo of how completely tannins, esp tannic acid, can convert a UrAc solution into a flocculent brown precipitate at the bottom of a UrAc-free supernatan (it LOOKS UrAc free! I made no measurements.). TA is cheap in non-EM grades; maybe they'd accept precipitation as a way of rendering it safely bio-inactive. From the information online at EMS ib their datasheet/22400, I've estimated that the 10,400 counts/secin depleted uranium is reduced to about -2 ct/sec in a very small bundle of muscle fibers containing 1 ug of protein and binding maybe 5 ug of UrAc. But it still requires health police and special containment if such a specimen is to be transported into or within the DOE facility at Argonne National Laboratory. The inconvenience has so far discouraged me from pursuing some experiments that would require such a specimen, but one day I will do it, legally and according to their regulations. I doubt a Geiger counter could detect any rise above background in the presence of that small an amount of UrAc.
-mike-
} Thanks for this Mike. I think KMnO4 followed by Sato's is one of the } better alternatives I've seen so far. Your results are very impressive. } } } Any idea if KMnO4 will work with LR White embedded tissues? } } Thanks, } Jessica Cervantes } Bend Research Inc } } -----Original Message----- } From: Mike Reedy [mailto:mike.reedy-at-cellbio.duke.edu] } Sent: Saturday, April 14, 2007 5:19 PM } To: Cervantes, Jessica } Subject: Re: [Microscopy] TEM: Alternative to Uranyl Acetate Stain } } You will get even more contrast if you use KMnO4 followed by Sato's, } as we have since 1964 (when Pb was was Pb cittrate, not Sato's). } And, you can get good contrast on thinner sections. Using Tannic } acid in the block stain, before OSO4, or after but followed by UrAc, } adds to contrast, improves preservation. We have had excellent } results (see attached). } } Some reports of using TA as a section stain before Pb stain can also } be found with google help. I've never tried it yet. } } KMnO4 section stain won't work if NMA is in your Epon mix. see } attached. } -mike reedy- } } ----------------------------------------------------------------------- } ----- } } 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
That would be a good place to start. I have posted some quite images of inclusions in an organic goo on our web site (ftp://www.marl.iastate.edu/Interesting/Residue/) . The mineral inclusions show up nicely.
Another poster mentioned doing this on a carbon substrate. In fact, I prepared this sample twice - once on a carbon stub and once on an aluminum stub. I wanted to see how much signal was coming from below. That was mostly for EDS and did appreciably affect the images. If you had a mixture of particles only, the situation might be a little different and I would recommend the dark background of a carbon substrate.
You may run up against resolution limits for BSE depending on particle size. The images may not be particularly sharp due to the sizeable interaction volume and the high currents typically required for BSE. These images were collected at 25mm WD with a sizeable current for simultaneous EDS. You could improve resolution by cutting the working distance and reducing current as much as possible while still maintaining signal strength.
Warren Straszheim Materials Analysis and Research Lab Iowa State University
-----Original Message----- X-from: nizets2-at-yahoo.com [mailto:nizets2-at-yahoo.com] Sent: Wednesday, April 18, 2007 8:05 AM To: wesaia-at-iastate.edu
Fellow Microscopists,
A few days ago I posted a query for suggestions on how to stabilize lysosomal membranes in cultured cells prior to immunocytochemistry. We have tried several of the tips but without success. Thanks to those who replied! We have been following the condition of the lysosomes using a GFP tag that we know to be specifically compartmentalized. We note that following fixation and a rinse in TRIS-Glycine the GFP is still compartmentalized. However, perhaps in 90% EtOH and definitely in 1:1 LR White:90% EtOH we see the GFP moves from the lysosomes to the cytoplasm and nucleus. It seems that ethanol and certainly LR White contribute to the deterioration of membranes. Can anyone suggest an embedding media that might be a little gentler on membranes and still possibly work for surface label immunocytochemistry? We are thinking of trying Nanoplast, a water soluble media but any suggestions are welcome.
Thanks! Doug
Douglas R. Keene Assistant Investigator Micro-Imaging Center Shriners Hospital for Children 3102 S.W. Sam Jackson Park Road Portland, Oregon 97239 503-221-3434 drk-at-shcc.org
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We get nice contrast with 1% potassium permanganate (aq) followed by lead citrate. It is good for membranes. In the past we have made it up in 0.1M phosphate buffer at pH below 6.5 to avoid precipitates.
Dave
-----Original Message----- X-from: cervantes-at-bendres.com [mailto:cervantes-at-bendres.com] Sent: 13 April 2007 21:16 To: David Patton
Due to some bureaucratic mumbo-jumbo, I am not able to use Uranyl Acetate (nuclear warheads, blah, blah, blah). I would like to stain some osmium-fixed tissue thin-sections for TEM; the procedure I'm following has a UA/Sato's lead stain step for the sections. Does anyone know of a suitable alternative? I did a quick google and listserver archive search and didn't find anything, but I'm hoping someone has run into this problem before and can suggest something.
Crossing my fingers, Jessica Cervantes Bend Research, Inc Bend, OR
==============================Original Headers============================== 3, 16 -- From cervantes-at-bendres.com Fri Apr 13 15:12:47 2007 3, 16 -- Received: from BRIEX04A.bri.local (bc161112.bendcable.com [216.228.161.112]) 3, 16 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3DKClG6024466 3, 16 -- for {Microscopy-at-microscopy.com} ; Fri, 13 Apr 2007 15:12:47 -0500 3, 16 -- MIME-Version: 1.0 3, 16 -- Content-Type: text/plain; 3, 16 -- charset="us-ascii" 3, 16 -- Subject: TEM: Alternative to Uranyl Acetate Stain 3, 16 -- Date: Fri, 13 Apr 2007 13:12:46 -0700 3, 16 -- Message-ID: {8943D65F9AD70E4488AD6DE09F15088768C44D-at-BRIEX04A} 3, 16 -- In-Reply-To: {8943D65F9AD70E4488AD6DE09F15088768C44C-at-BRIEX04A} 3, 16 -- References: {8943D65F9AD70E4488AD6DE09F15088768C44C-at-BRIEX04A} 3, 16 -- From: "Cervantes, Jessica" {cervantes-at-bendres.com} 3, 16 -- To: {Microscopy-at-microscopy.com} 3, 16 -- Content-Transfer-Encoding: 8bit 3, 16 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l3DKClG6024466 ==============================End of - Headers==============================
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==============================Original Headers============================== 15, 34 -- From David.Patton-at-uwe.ac.uk Thu Apr 19 05:29:36 2007 15, 34 -- Received: from mailapp04.uwe.ac.uk (mailapp04.uwe.ac.uk [164.11.132.66]) 15, 34 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id l3JATZZM014188 15, 34 -- for {Microscopy-at-microscopy.com} ; Thu, 19 Apr 2007 05:29:36 -0500 15, 34 -- Received: from (mta01.uwe.ac.uk [164.11.132.60]) by mailapp04.uwe.ac.uk with smtp 15, 34 -- id 77fc_85ccbd46_ee60_11db_9c4e_00142223915c; 15, 34 -- Thu, 19 Apr 2007 11:27:19 +0100 15, 34 -- Received: from egen-uwe01.campus.ads.uwe.ac.uk 15, 34 -- (egen-uwe01.uwe.ac.uk [164.11.249.121]) 15, 34 -- by mta01.uwe.ac.uk (iPlanet Messaging Server 5.2 HotFix 2.07 (built Jun 24 15, 34 -- 2005)) with ESMTP id {0JGQ00G8RQGVVP-at-mta01.uwe.ac.uk} for 15, 34 -- Microscopy-at-microscopy.com; Thu, 19 Apr 2007 11:29:20 +0100 (BST) 15, 34 -- Date: Thu, 19 Apr 2007 11:25:30 +0100 15, 34 -- From: David Patton {David.Patton-at-uwe.ac.uk} 15, 34 -- Subject: RE: [Microscopy] TEM: Alternative to Uranyl Acetate Stain 15, 34 -- In-reply-to: {200704132016.l3DKG8er031358-at-ns.microscopy.com} 15, 34 -- To: cervantes-at-bendres.com 15, 34 -- Cc: Microscopy-at-microscopy.com 15, 34 -- Message-id: {F247F674896BE243AD8263C5280E2BDB02D910E2-at-egen-uwe01} 15, 34 -- MIME-version: 1.0 15, 34 -- X-MIMEOLE: Produced By Microsoft Exchange V6.5 15, 34 -- Content-type: text/plain; charset=us-ascii 15, 34 -- Content-class: urn:content-classes:message 15, 34 -- Thread-topic: [Microscopy] TEM: Alternative to Uranyl Acetate Stain 15, 34 -- Thread-index: Acd+CJpt4UbiUWZfTSeZjlib8dd1eAEY27Gw 15, 34 -- X-MS-Has-Attach: 15, 34 -- X-MS-TNEF-Correlator: 15, 34 -- X-NAIMIME-Disclaimer: 1 15, 34 -- X-NAIMIME-Modified: 1 15, 34 -- X-NAI-Spam-Score: -1.2 15, 34 -- X-NAI-Spam-Rules: 1 Rules triggered 15, 34 -- BAYES_01=-1.2 15, 34 -- Content-Transfer-Encoding: 8bit 15, 34 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l3JATZZM014188 ==============================End of - Headers==============================
Sorry - just back from leave! Recently we have used it on 20yr old epoxy blocks of unknown provenance and TAAB embedding resin. No experience on LR White.
Will try Pal's bleach as we seem to have some non-specific staining.
It was in use here in 1989, when I started here, due to a safety scare (yes even back in the good old days!). Hayat (1989) noted precipitates above pH 6.8. We have never investigated the maintainance of pH. We stain for 5-10min with KMnO4.
Dave
-----Original Message----- X-from: Mike Reedy [mailto:mike.reedy-at-cellbio.duke.edu] Sent: 19 April 2007 15:20 To: David Patton Cc: cervantes-at-bendres.com
-- -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
Here's the list of references. I haven't checked if all are available or still in print.
I'm replying to the whole list in case anyone else is interested (can't tell if you sent your message just to me).
Jessica Cervantes Bend Research Inc Bend, OR
The Cell, by Don W. Fawcett, MD. W. B. Saunders Company (out of print).
Biomedical Electron Microscopy - Illustrated Methods and Interpretations, by Arvid B. Maunsbach and Bjorn A. Afzelius. Academic Press; 1st edition (January 15, 1999).
Cell and tissue ultrastructure: a functional perspective, by P. C. Cross, and K. L. Mercer. W. H. Freeman & Co, 1993. 420 pp.
Histology: A text and Atlas, by Michael H. Ross, and Wojciech Pawlina. Lippincott Williams & Wilkins; 1st edition (December 1, 2005). Was updated 2006 (paperback) with CD-ROM and is available from Barnes & Noble.
Color Atlas of Cytology, Histology and Microscopic Anatomy, by Wolfgang Kuehnel. Thieme Medical Publishers; 4th edition (July 2003).
An Atlas of Ultrastructure, by Johannes Rhodin. W.B. Saunders Co.
Bloom and Fawcett: Concise Histology, by Don W. Fawcett, Ronald P. Jensh. A Hodder Arnold Publication; 2nd edition (June 15, 2002).
For plant cells: Introduction to the Fine Structure of Plant Cells, Myron Ledbetter and Keith Porter. Springer-Verlag.
-----Original Message----- X-from: Prof. dr. Wim Van den Broeck [mailto:wim.vandenbroeck-at-UGent.be] Sent: Thursday, April 19, 2007 12:29 AM To: Cervantes, Jessica
It seems that I've just been gifted with an LKB Nova ultratome, complete with hydrolic table. However, there are no operator's manuals with the microtome. Set it up and it is working fine, but would really like to have an operator's manual so that I can check and make sure we know all the little ins and outs of the machine.
Can any one help?????
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
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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 scott.streiker-at-udri.udayton.edu as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: scott.streiker-at-udri.udayton.edu Name: Scott Streiker
Organization: University of Dayton Research Institute
Title-Subject: [Filtered] Uptake of nanoparticles in cells using TEM
Question: Hello! I am trying to visualize the uptake of nanomaterials in cells using TEM. I have a cell pellet that I would like to embed in resin and then section using the ultramicrotome. One kit that I have the option to use is the Epofix Cold-Setting Resin from EMS. Has anyone used this kit before on biological samples? Or is there a better resin to try?
For plant cells there are also books by Brian Gunning
Brian E.S Gunning and Martin W. Steer 'Plant Cell Biology; Structure and Function' Jones and Bartlett Publishers (1996)
Brian E.S Gunning and Martin W. Steer 'Ultrastructure and the Biology of Plant Cells' Edward Arnold (older version from mid 1970's
There is also a new DVD due out mid 2007 (I have seen an early version which looks very good). You can find information at:www.plantcellbiologyondvd.com/default.cfm
Ian
Ian Hallett Sensory and Consumer Science - Microscopy HortResearch, Mt Albert Research Centre Private Bag 92 169, Auckland Mail Centre Auckland 1142, New Zealand +64-9-815 4200 ext 7002
-----Original Message----- X-from: cervantes-at-bendres.com [mailto:cervantes-at-bendres.com] Sent: Friday, 20 April 2007 3:35 a.m. To: Ian Hallett
Wim -
Here's the list of references. I haven't checked if all are available or still in print.
I'm replying to the whole list in case anyone else is interested (can't tell if you sent your message just to me).
Jessica Cervantes Bend Research Inc Bend, OR
The Cell, by Don W. Fawcett, MD. W. B. Saunders Company (out of print).
Biomedical Electron Microscopy - Illustrated Methods and Interpretations, by Arvid B. Maunsbach and Bjorn A. Afzelius. Academic Press; 1st edition (January 15, 1999).
Cell and tissue ultrastructure: a functional perspective, by P. C. Cross, and K. L. Mercer. W. H. Freeman & Co, 1993. 420 pp.
Histology: A text and Atlas, by Michael H. Ross, and Wojciech Pawlina. Lippincott Williams & Wilkins; 1st edition (December 1, 2005). Was updated 2006 (paperback) with CD-ROM and is available from Barnes & Noble.
Color Atlas of Cytology, Histology and Microscopic Anatomy, by Wolfgang Kuehnel. Thieme Medical Publishers; 4th edition (July 2003).
An Atlas of Ultrastructure, by Johannes Rhodin. W.B. Saunders Co.
Bloom and Fawcett: Concise Histology, by Don W. Fawcett, Ronald P. Jensh. A Hodder Arnold Publication; 2nd edition (June 15, 2002).
For plant cells: Introduction to the Fine Structure of Plant Cells, Myron Ledbetter and Keith Porter. Springer-Verlag.
-----Original Message----- X-from: Prof. dr. Wim Van den Broeck [mailto:wim.vandenbroeck-at-UGent.be] Sent: Thursday, April 19, 2007 12:29 AM To: Cervantes, Jessica
Hi, Scott
This would definitely be another of those interesting applications to try with the new Tomo AFM/Ultramicrotome from NT-MDT. The AFM is superb at imaging nanoparticles and TOMO uses the Leica ultramicrotome for sectioning.
While I don't have any pictures of nanoparticles, I do have a really neat movie I can share with you showing the serial sectioning of nanotubes in epoxy, followed by the dynamic 3D reconstruction from Dr. DeWith at the Dutch Polymer Institute at the TU/Eindhoven. Contact me off-line if you are interested. I can send it via YouSendIt so that you can download it easily. Also, if you are interested in seeing if this is a good solution for your application, I encourage you to correspond directly with Dr. Efimov at NTMDT (see CC above). He may be able to run a sample for you.
Hope this is helpful.
Best regards, Barbara Foster
Microscopy/Microscopy Education 313 S Jupiter Rd, Suite 100 Allen, TX 75002 P: 972-954-8011 W: www.MicroscopyEducation.com
Caveat: MME is working with NTMDT in support of the TOMO.
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 04:16 PM 4/19/2007, scott.streiker-at-udri.udayton.edu wrote:
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==============================Original Headers============================== 15, 20 -- From bfoster-at-mme1.com Thu Apr 19 16:44:54 2007 15, 20 -- Received: from smtp109.sbc.mail.mud.yahoo.com (smtp109.sbc.mail.mud.yahoo.com [68.142.198.208]) 15, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id l3JLisjA031270 15, 20 -- for {microscopy-at-microscopy.com} ; Thu, 19 Apr 2007 16:44:54 -0500 15, 20 -- Message-Id: {200704192144.l3JLisjA031270-at-ns.microscopy.com} 15, 20 -- Received: (qmail 6361 invoked from network); 19 Apr 2007 21:44:54 -0000 15, 20 -- Received: from unknown (HELO barbsd505.mme1.com) (bfostermme-at-sbcglobal.net-at-68.94.41.65 with login) 15, 20 -- by smtp109.sbc.mail.mud.yahoo.com with SMTP; 19 Apr 2007 21:44:53 -0000 15, 20 -- X-YMail-OSG: HdTn93gVM1mo7bTZPN6PMkttZnZtL7CmrzG8WxRvU5Q_V9AFKgFSPXo0HIGqJqI5KDJ8Fwsji4u_AlXS6x1.CRr7rBLeB.YuNG8eKLFLZ31L0hctbs6G_QufjSeuQkpzbLtZXECQ4pdnw21XGxVpEnnemRo- 15, 20 -- X-Mailer: QUALCOMM Windows Eudora Version 7.1.0.9 15, 20 -- Date: Thu, 19 Apr 2007 16:44:26 -0500 15, 20 -- To: scott.streiker-at-udri.udayton.edu, microscopy-at-microscopy.com 15, 20 -- From: Barbara Foster {bfoster-at-mme1.com} 15, 20 -- Subject: Re: [Microscopy] viaWWW: Uptake of nanoparticles in cells 15, 20 -- using TEM 15, 20 -- Cc: Anton Efimov {efimov-at-ntmdt.ru} 15, 20 -- In-Reply-To: {200704191944.l3JJi9ij010448-at-ns.microscopy.com} 15, 20 -- References: {200704191944.l3JJi9ij010448-at-ns.microscopy.com} 15, 20 -- Mime-Version: 1.0 15, 20 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
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Email: eknoppel-at-cc.usu.edu Name: Edward L. Knoppel
Organization: USDA-ARS-PPRL
Title-Subject: [Filtered] Lung tissue
Question: Without using a microwave, freezing or perfusion of the tissue; is there a "really" good procedure for fixing pieces of animal lung tissue that someone would be gracious enough to share? Thanks in advance, Ed
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Bruker AXS, Inc., a leading manufacturer of Analytical Instrumentation, has a job opening for a Senior Salesperson, working out of the Bruker Canada office in Milton, Ontario.
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Did I miss the replies to this query somehow? Is there a technical problem, or were the replies private? Would it be possible to post them?
Let's not be shy (if that's it) about posting to the whole list. I've gotten so much out of this resource (especially lately). I've had to re-post replies to my query for others who have had the same question and hadn't gotten the messages.
Thanks for keeping the information flowing; it's invaluable. Jessica Cervantes Bend Research Inc Bend, OR
-----Original Message----- X-from: drk-at-SHCC.org [mailto:drk-at-SHCC.org] Sent: Wednesday, April 18, 2007 10:49 AM To: Cervantes, Jessica
Fellow Microscopists,
A few days ago I posted a query for suggestions on how to stabilize lysosomal membranes in cultured cells prior to immunocytochemistry. We have tried several of the tips but without success. Thanks to those who replied! We have been following the condition of the lysosomes using a GFP tag that we know to be specifically compartmentalized. We note that following fixation and a rinse in TRIS-Glycine the GFP is still compartmentalized. However, perhaps in 90% EtOH and definitely in 1:1 LR White:90% EtOH we see the GFP moves from the lysosomes to the cytoplasm and nucleus. It seems that ethanol and certainly LR White contribute to the deterioration of membranes. Can anyone suggest an embedding media that might be a little gentler on membranes and still possibly work for surface label immunocytochemistry? We are thinking of trying Nanoplast, a water soluble media but any suggestions are welcome.
Thanks! Doug
Douglas R. Keene Assistant Investigator Micro-Imaging Center Shriners Hospital for Children 3102 S.W. Sam Jackson Park Road Portland, Oregon 97239 503-221-3434 drk-at-shcc.org
==============================Original Headers============================== 7, 22 -- From drk-at-SHCC.org Wed Apr 18 12:43:18 2007 7, 22 -- Received: from mail.SHCC.org (mail.shcc.org [64.213.211.202]) 7, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3IHhITP002098 7, 22 -- for {Microscopy-at-Microscopy.Com} ; Wed, 18 Apr 2007 12:43:18 -0500 7, 22 -- Received: from DRK2 ([64.213.211.62]) by mail.SHCC.org (PMDF V6.3 #31473) 7, 22 -- with ESMTPA id {0JGP0029FFTXS8-at-mail.SHCC.org} for Microscopy-at-Microscopy.Com; 7, 22 -- Wed, 18 Apr 2007 10:41:57 -0700 (PDT) 7, 22 -- Date: Wed, 18 Apr 2007 10:43:58 -0700 7, 22 -- From: Doug Keene {drk-at-SHCC.org} 7, 22 -- Subject: stabilization ofmembranes 7, 22 -- Sender: drk-at-SHCC.org 7, 22 -- To: Microscopy-at-Microscopy.Com 7, 22 -- Cc: drk-at-SHCC.org 7, 22 -- Reply-to: drk-at-SHCC.org 7, 22 -- Message-id: {0JGP0029GFTXS8-at-mail.SHCC.org} 7, 22 -- Organization: Shriners Hospitals for Children 7, 22 -- MIME-version: 1.0 7, 22 -- X-MIMEOLE: Produced By Microsoft MimeOLE V6.00.2900.3028 7, 22 -- X-Mailer: Microsoft Office Outlook, Build 11.0.5510 7, 22 -- Content-type: text/plain; charset=us-ascii 7, 22 -- Content-transfer-encoding: 7BIT 7, 22 -- Thread-Index: AceB4SRWHFc2k9QuQCCG4KbQ3H8v4A== ==============================End of - Headers==============================
==============================Original Headers============================== 13, 16 -- From cervantes-at-bendres.com Thu Apr 19 17:57:57 2007 13, 16 -- Received: from BRIEX04A.bri.local (bc161112.bendcable.com [216.228.161.112]) 13, 16 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3JMvuoK002012 13, 16 -- for {Microscopy-at-microscopy.com} ; Thu, 19 Apr 2007 17:57:56 -0500 13, 16 -- MIME-Version: 1.0 13, 16 -- Content-Type: text/plain; 13, 16 -- charset="us-ascii" 13, 16 -- Subject: RE: [Microscopy] stabilization of membranes 13, 16 -- Date: Thu, 19 Apr 2007 15:57:55 -0700 13, 16 -- Message-ID: {8943D65F9AD70E4488AD6DE09F15088768C47F-at-BRIEX04A} 13, 16 -- In-Reply-To: {200704181748.l3IHmsrE012175-at-ns.microscopy.com} 13, 16 -- References: {200704181748.l3IHmsrE012175-at-ns.microscopy.com} 13, 16 -- From: "Cervantes, Jessica" {cervantes-at-bendres.com} 13, 16 -- To: {Microscopy-at-microscopy.com} 13, 16 -- Content-Transfer-Encoding: 8bit 13, 16 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l3JMvuoK002012 ==============================End of - Headers==============================
I would like to 2nd the idea that replying to the list is a good thing. This is my only resource for such information so I would rather delete than miss an opportunity to learn something.
Thanks!
Tom Kaye
Not affiliated with anything.
-----Original Message----- X-from: cervantes-at-bendres.com [mailto:cervantes-at-bendres.com] Sent: Thursday, April 19, 2007 6:02 PM To: tom-at-tomkaye.com
Did I miss the replies to this query somehow? Is there a technical problem, or were the replies private? Would it be possible to post them?
Let's not be shy (if that's it) about posting to the whole list. I've gotten so much out of this resource (especially lately). I've had to re-post replies to my query for others who have had the same question and hadn't gotten the messages.
Thanks for keeping the information flowing; it's invaluable. Jessica Cervantes Bend Research Inc Bend, OR
-----Original Message----- X-from: drk-at-SHCC.org [mailto:drk-at-SHCC.org] Sent: Wednesday, April 18, 2007 10:49 AM To: Cervantes, Jessica
Fellow Microscopists,
A few days ago I posted a query for suggestions on how to stabilize lysosomal membranes in cultured cells prior to immunocytochemistry. We have tried several of the tips but without success. Thanks to those who replied! We have been following the condition of the lysosomes using a GFP tag that we know to be specifically compartmentalized. We note that following fixation and a rinse in TRIS-Glycine the GFP is still compartmentalized. However, perhaps in 90% EtOH and definitely in 1:1 LR White:90% EtOH we see the GFP moves from the lysosomes to the cytoplasm and nucleus. It seems that ethanol and certainly LR White contribute to the deterioration of membranes. Can anyone suggest an embedding media that might be a little gentler on membranes and still possibly work for surface label immunocytochemistry? We are thinking of trying Nanoplast, a water soluble media but any suggestions are welcome.
Thanks! Doug
Douglas R. Keene Assistant Investigator Micro-Imaging Center Shriners Hospital for Children 3102 S.W. Sam Jackson Park Road Portland, Oregon 97239 503-221-3434 drk-at-shcc.org
==============================Original Headers============================== 7, 22 -- From drk-at-SHCC.org Wed Apr 18 12:43:18 2007 7, 22 -- Received: from mail.SHCC.org (mail.shcc.org [64.213.211.202]) 7, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3IHhITP002098 7, 22 -- for {Microscopy-at-Microscopy.Com} ; Wed, 18 Apr 2007 12:43:18 -0500 7, 22 -- Received: from DRK2 ([64.213.211.62]) by mail.SHCC.org (PMDF V6.3 #31473) 7, 22 -- with ESMTPA id {0JGP0029FFTXS8-at-mail.SHCC.org} for Microscopy-at-Microscopy.Com; 7, 22 -- Wed, 18 Apr 2007 10:41:57 -0700 (PDT) 7, 22 -- Date: Wed, 18 Apr 2007 10:43:58 -0700 7, 22 -- From: Doug Keene {drk-at-SHCC.org} 7, 22 -- Subject: stabilization ofmembranes 7, 22 -- Sender: drk-at-SHCC.org 7, 22 -- To: Microscopy-at-Microscopy.Com 7, 22 -- Cc: drk-at-SHCC.org 7, 22 -- Reply-to: drk-at-SHCC.org 7, 22 -- Message-id: {0JGP0029GFTXS8-at-mail.SHCC.org} 7, 22 -- Organization: Shriners Hospitals for Children 7, 22 -- MIME-version: 1.0 7, 22 -- X-MIMEOLE: Produced By Microsoft MimeOLE V6.00.2900.3028 7, 22 -- X-Mailer: Microsoft Office Outlook, Build 11.0.5510 7, 22 -- Content-type: text/plain; charset=us-ascii 7, 22 -- Content-transfer-encoding: 7BIT 7, 22 -- Thread-Index: AceB4SRWHFc2k9QuQCCG4KbQ3H8v4A== ==============================End of - Headers==============================
==============================Original Headers============================== 13, 16 -- From cervantes-at-bendres.com Thu Apr 19 17:57:57 2007 13, 16 -- Received: from BRIEX04A.bri.local (bc161112.bendcable.com [216.228.161.112]) 13, 16 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3JMvuoK002012 13, 16 -- for {Microscopy-at-microscopy.com} ; Thu, 19 Apr 2007 17:57:56 -0500 13, 16 -- MIME-Version: 1.0 13, 16 -- Content-Type: text/plain; 13, 16 -- charset="us-ascii" 13, 16 -- Subject: RE: [Microscopy] stabilization of membranes 13, 16 -- Date: Thu, 19 Apr 2007 15:57:55 -0700 13, 16 -- Message-ID: {8943D65F9AD70E4488AD6DE09F15088768C47F-at-BRIEX04A} 13, 16 -- In-Reply-To: {200704181748.l3IHmsrE012175-at-ns.microscopy.com} 13, 16 -- References: {200704181748.l3IHmsrE012175-at-ns.microscopy.com} 13, 16 -- From: "Cervantes, Jessica" {cervantes-at-bendres.com} 13, 16 -- To: {Microscopy-at-microscopy.com} 13, 16 -- Content-Transfer-Encoding: 8bit 13, 16 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l3JMvuoK002012 ==============================End of - Headers==============================
==============================Original Headers============================== 25, 20 -- From tom-at-tomkaye.com Thu Apr 19 18:47:51 2007 25, 20 -- Received: from tomkaye.com (mxgateway.techpro.com [64.4.207.8]) 25, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3JNlodW014261 25, 20 -- for {microscopy-at-microscopy.com} ; Thu, 19 Apr 2007 18:47:51 -0500 25, 20 -- Received: from tk7c797a275194 [24.15.58.103] by tomkaye.com with ESMTP 25, 20 -- (SMTPD32-8.14) id AFAD3312012A; Thu, 19 Apr 2007 18:47:57 -0500 25, 20 -- From: "Tom Kaye" {tom-at-tomkaye.com} 25, 20 -- To: {microscopy-at-microscopy.com} 25, 20 -- Subject: 2nd for posting to the list 25, 20 -- Date: Thu, 19 Apr 2007 18:48:03 -0500 25, 20 -- Message-ID: {GOEAIJOJDGCJBPIFHOBGKEPBOOAA.tom-at-tomkaye.com} 25, 20 -- MIME-Version: 1.0 25, 20 -- Content-Type: text/plain; 25, 20 -- charset="iso-8859-1" 25, 20 -- Content-Transfer-Encoding: 7bit 25, 20 -- X-Priority: 3 (Normal) 25, 20 -- X-MSMail-Priority: Normal 25, 20 -- X-Mailer: Microsoft Outlook IMO, Build 9.0.2416 (9.0.2911.0) 25, 20 -- x-mimeole: Produced By Microsoft MimeOLE V6.00.2900.3028 25, 20 -- Importance: Normal ==============================End of - Headers==============================
Do any of you know what this thing is? It looks like a "bug" but it's attached to a probable piece of fungus. Any ideas welcome, this is a head scratcher for everyone that has seen it.
Thanks!
Tom Kaye
Tomkaye.com/images/fungus_lrg.jpg Long shot, the bug thing is lower right
Tomkaye.com/images/fungus2_lrg.jpg Close up.
==============================Original Headers============================== 8, 20 -- From tom-at-tomkaye.com Thu Apr 19 19:07:05 2007 8, 20 -- Received: from tomkaye.com (mxgateway.techpro.com [64.4.207.8]) 8, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3K075vP025985 8, 20 -- for {microscopy-at-microscopy.com} ; Thu, 19 Apr 2007 19:07:05 -0500 8, 20 -- Received: from tk7c797a275194 [24.15.58.103] by tomkaye.com with ESMTP 8, 20 -- (SMTPD32-8.14) id A4303503012A; Thu, 19 Apr 2007 19:07:12 -0500 8, 20 -- From: "Tom Kaye" {tom-at-tomkaye.com} 8, 20 -- To: {microscopy-at-microscopy.com} 8, 20 -- Subject: Need help with ID 8, 20 -- Date: Thu, 19 Apr 2007 19:07:18 -0500 8, 20 -- Message-ID: {GOEAIJOJDGCJBPIFHOBGCEPEOOAA.tom-at-tomkaye.com} 8, 20 -- MIME-Version: 1.0 8, 20 -- Content-Type: text/plain; 8, 20 -- charset="iso-8859-1" 8, 20 -- Content-Transfer-Encoding: 7bit 8, 20 -- X-Priority: 3 (Normal) 8, 20 -- X-MSMail-Priority: Normal 8, 20 -- X-Mailer: Microsoft Outlook IMO, Build 9.0.2416 (9.0.2911.0) 8, 20 -- x-mimeole: Produced By Microsoft MimeOLE V6.00.2900.3028 8, 20 -- Importance: Normal ==============================End of - Headers==============================
Hello Ed, There is a method which works really well in fetal mouse lung. (Cole TJ, Solomon NM, van Driel R, Monk JA, Bird D, Richardson SJ, Dilley RJ, Hooper SB. Am J Respir Cell Mol Biol 2004 May; 30(5):613-9 "Altered epithelial proportions in the fetal lung of glucocorticoid receptor null mice")
We used the method as published by Williams, M. C. 1977. "Conversion of lamellar body membranes into tubular myelin in alveoli of fetal rat lungs". J. Cell Biol. 72:260-277.
It involves use of veronal (barbitone) and maleate buffers, but is really simple to do.
However, I do not know how well it works with inflated lungs.
Briefly: Dissect lungs from embryo, place in drop of fixative and slice gently into mm cubes. Fix in 4% Paraformaldehyde + 2% Glutaraldehyde + 4% Sucrose in HEPES buffered saline pH 7.4 for 3 to 4 hours at room temp. Rinse 2 min in cold veronal acetate pH 7.4 Postfix in 1.5% OsO4 in Veronal Acetate -at- 4degrees C overnight Rinse 3 x 10 min -at-4 deg C in Tris Maleate pH 5.2 En bloc stain with 1.5% UrAc in Tris Maleate pH5.2, 90 min on ice, in dark Cold dehydration, 5 minute changes in graded acetones on ice (10, 20, 30, 40, 50, 60, 70, 80, 90, 95%)Then Absolute dry acetone 4 x 10 minutes, the last 2 changes at room temp. Absolute acetone:Epon Mix, 50:50, 30 minutes, rotating Epon, 3 x 60 minute changes, can leave one change overnight, rotating Embed in fresh Epon, polymerize overnight at 60 to 65 degrees C.
If you would like the buffer recipes, let me know.
Good luck!
Rosey
Rosemary van Driel Electron Microscopy New and Emerging Zoonotic Diseases Australian Animal Health Laboratory CSIRO Livestock Industries Private Bag 24 Geelong Vic 3220 Australia
International Phone: +61 3 5227 5209 International Fax: +61 3 5227 5555 email: Rosey.VanDriel-at-csiro.au
-----Original Message----- X-from: eknoppel-at-cc.usu.edu [mailto:eknoppel-at-cc.usu.edu] Sent: Friday, 20 April 2007 08:00 To: Van Driel, Rosey (LI, Geelong)
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 eknoppel-at-cc.usu.edu as well as the MIcroscopy Listserver ------------------------------------------------------------------------ ---
Email: eknoppel-at-cc.usu.edu Name: Edward L. Knoppel
Organization: USDA-ARS-PPRL
Title-Subject: [Filtered] Lung tissue
Question: Without using a microwave, freezing or perfusion of the tissue; is there a "really" good procedure for fixing pieces of animal lung tissue that someone would be gracious enough to share? Thanks in advance, Ed
Just a guess - looks like the moulted exoskeleton of a caterpillar-like insect larva. The nice wavy tubes look like the air circulation tubes that connect to spiracles.
Dave
-----Original Message----- X-from: tom-at-tomkaye.com [mailto:tom-at-tomkaye.com] Sent: 20 April 2007 01:11 To: David Patton
Hello All,
Do any of you know what this thing is? It looks like a "bug" but it's attached to a probable piece of fungus. Any ideas welcome, this is a head scratcher for everyone that has seen it.
Thanks!
Tom Kaye
Tomkaye.com/images/fungus_lrg.jpg Long shot, the bug thing is lower right
Tomkaye.com/images/fungus2_lrg.jpg Close up.
==============================Original Headers============================== 8, 20 -- From tom-at-tomkaye.com Thu Apr 19 19:07:05 2007 8, 20 -- Received: from tomkaye.com (mxgateway.techpro.com [64.4.207.8]) 8, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3K075vP025985 8, 20 -- for {microscopy-at-microscopy.com} ; Thu, 19 Apr 2007 19:07:05 -0500 8, 20 -- Received: from tk7c797a275194 [24.15.58.103] by tomkaye.com with ESMTP 8, 20 -- (SMTPD32-8.14) id A4303503012A; Thu, 19 Apr 2007 19:07:12 -0500 8, 20 -- From: "Tom Kaye" {tom-at-tomkaye.com} 8, 20 -- To: {microscopy-at-microscopy.com} 8, 20 -- Subject: Need help with ID 8, 20 -- Date: Thu, 19 Apr 2007 19:07:18 -0500 8, 20 -- Message-ID: {GOEAIJOJDGCJBPIFHOBGCEPEOOAA.tom-at-tomkaye.com} 8, 20 -- MIME-Version: 1.0 8, 20 -- Content-Type: text/plain; 8, 20 -- charset="iso-8859-1" 8, 20 -- Content-Transfer-Encoding: 7bit 8, 20 -- X-Priority: 3 (Normal) 8, 20 -- X-MSMail-Priority: Normal 8, 20 -- X-Mailer: Microsoft Outlook IMO, Build 9.0.2416 (9.0.2911.0) 8, 20 -- x-mimeole: Produced By Microsoft MimeOLE V6.00.2900.3028 8, 20 -- Importance: Normal ==============================End of - Headers==============================
This incoming email to UWE has been independently scanned for viruses by McAfee anti-virus software and none were detected
This email was independently scanned for viruses by McAfee anti-virus software and none were found
==============================Original Headers============================== 20, 34 -- From David.Patton-at-uwe.ac.uk Fri Apr 20 03:49:14 2007 20, 34 -- Received: from mailapp04.uwe.ac.uk (mailapp04.uwe.ac.uk [164.11.132.66]) 20, 34 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id l3K8nBlm025828 20, 34 -- for {microscopy-at-microscopy.com} ; Fri, 20 Apr 2007 03:49:13 -0500 20, 34 -- Received: from (mta01.uwe.ac.uk [164.11.132.60]) by mailapp04.uwe.ac.uk with smtp 20, 34 -- id 49da_aa568f1e_ef1b_11db_8139_00142223915c; 20, 34 -- Fri, 20 Apr 2007 09:46:53 +0100 20, 34 -- Received: from egen-uwe01.campus.ads.uwe.ac.uk 20, 34 -- (egen-uwe01.uwe.ac.uk [164.11.249.121]) 20, 34 -- by mta01.uwe.ac.uk (iPlanet Messaging Server 5.2 HotFix 2.07 (built Jun 24 20, 34 -- 2005)) with ESMTP id {0JGS00DSQGHNCO-at-mta01.uwe.ac.uk} for 20, 34 -- microscopy-at-microscopy.com; Fri, 20 Apr 2007 09:48:59 +0100 (BST) 20, 34 -- Date: Fri, 20 Apr 2007 09:47:51 +0100 20, 34 -- From: David Patton {David.Patton-at-uwe.ac.uk} 20, 34 -- Subject: RE: [Microscopy] Need help with ID 20, 34 -- In-reply-to: {200704200010.l3K0AdQj000362-at-ns.microscopy.com} 20, 34 -- To: tom-at-tomkaye.com 20, 34 -- Cc: microscopy-at-microscopy.com 20, 34 -- Message-id: {F247F674896BE243AD8263C5280E2BDB02D91120-at-egen-uwe01} 20, 34 -- MIME-version: 1.0 20, 34 -- X-MIMEOLE: Produced By Microsoft Exchange V6.5 20, 34 -- Content-type: text/plain; charset=us-ascii 20, 34 -- Content-class: urn:content-classes:message 20, 34 -- Thread-topic: [Microscopy] Need help with ID 20, 34 -- Thread-index: AceC4FstefBqksnjQ3mf/z7xkxVM3QAR6WhA 20, 34 -- X-MS-Has-Attach: 20, 34 -- X-MS-TNEF-Correlator: 20, 34 -- X-NAIMIME-Disclaimer: 1 20, 34 -- X-NAIMIME-Modified: 1 20, 34 -- X-NAI-Spam-Score: 0 20, 34 -- X-NAI-Spam-Rules: 1 Rules triggered 20, 34 -- BAYES_44=-0 20, 34 -- Content-Transfer-Encoding: 8bit 20, 34 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l3K8nBlm025828 ==============================End of - Headers==============================
I'm doing basic cross section preparation for light microscopy work. My samples are stainless steel plated with 0-50 micrometers thick layers of copper and chromium.
I would like to make the interfaces clearer than they are after grinding/polishing. I have thought about etching/corroding the surface with some acid or other chemical.
I would like to hear from the experts what kind of chemicals they might recommend. Or if etching my samples chemically would do me any good at all.
Thanks in advance
Niko Hellstén Product Engineer Stratum Oy mail: niko.hellsten-at-stratum.fi GSM: +358-(0)440-955301
==============================Original Headers============================== 7, 22 -- From niko.hellsten-at-stratum.fi Fri Apr 20 04:50:21 2007 7, 22 -- Received: from emh03.mail.saunalahti.fi (emh03.mail.saunalahti.fi [62.142.5.109]) 7, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3K9oK1j006613 7, 22 -- for {Microscopy-at-Microscopy.com} ; Fri, 20 Apr 2007 04:50:20 -0500 7, 22 -- Received: from Evkan (unknown [195.255.124.215]) 7, 22 -- by emh03.mail.saunalahti.fi (Postfix) with SMTP id 341A5158C6C 7, 22 -- for {Microscopy-at-Microscopy.com} ; Fri, 20 Apr 2007 12:50:18 +0300 (EEST) 7, 22 -- Message-ID: {002f01c78331$57183970$d77cffc3-at-stratum.fi} 7, 22 -- From: "Niko Hellsten" {niko.hellsten-at-stratum.fi} 7, 22 -- To: "Microscopy" {Microscopy-at-Microscopy.com} 7, 22 -- Subject: cross section sample preparation 7, 22 -- Date: Fri, 20 Apr 2007 12:50:26 +0300 7, 22 -- MIME-Version: 1.0 7, 22 -- Content-Type: text/plain; 7, 22 -- format=flowed; 7, 22 -- charset="iso-8859-1"; 7, 22 -- reply-type=original 7, 22 -- Content-Transfer-Encoding: 8bit 7, 22 -- X-Priority: 3 7, 22 -- X-MSMail-Priority: Normal 7, 22 -- X-Mailer: Microsoft Outlook Express 6.00.2900.3028 7, 22 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.3028 ==============================End of - Headers==============================
I do not know if it would be appropriate with your material but I always suggest people fracture samples for cross section. The usual method is to use liquid nitrogen to embrittle the specimen.
This method is mainly used for the SEM but many clients use LM as well. The SEM is very good at detecting poor cross sections through cutting or polishing but the fracture route has never let me down.
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: {niko.hellsten-at-stratum.fi} To: {protrain-at-emcourses.com} Sent: Friday, April 20, 2007 10:51 AM
Hello Niko,
If you are just trying to delineate the interfaces, I would etch the copper layer. This is assuming that your configuration is the SS base metal / a Cu under layer / a Cr outer layer. Of more importance is the polishing of the sample before etching. You must ensure that the polishing method does not introduce smearing of the polished surface, especially at the copper plating.
A flat etch to remove copper and reveal the copper grain boundaries consists of;
20 ml ammonium hydroxide 20 ml of water 5 ml of 3% hydrogen peroxide
The hydrogen peroxide is added just prior to etching. Etching is performed by swabbing the polished surface for approximately 10-30 sec.
For more on polishing methods see;
"ASM Handbook, Vol. 9 - Metallography and Microstructures", ASM International, 2004, Materials Park, OH 44073
For etchants and precautions for safely using them see;
Joseph M. Oparowski Center for Materials Science Bose Corporation Joseph_Oparowski-at-bose.com The Mountain, M/S 415 Phone: (508) 766-1371 Framingham, MA 01701-9168 Fax: (508) 766-1313
-----Original Message----- X-from: niko.hellsten-at-stratum.fi [mailto:niko.hellsten-at-stratum.fi] Sent: Friday, April 20, 2007 5:55 AM To: Oparowski, Joseph
Hello everyone
I'm doing basic cross section preparation for light microscopy work. My samples are stainless steel plated with 0-50 micrometers thick layers of copper and chromium.
I would like to make the interfaces clearer than they are after grinding/polishing. I have thought about etching/corroding the surface with some acid or other chemical.
I would like to hear from the experts what kind of chemicals they might recommend. Or if etching my samples chemically would do me any good at all.
Thanks in advance
Niko Hellstén Product Engineer Stratum Oy mail: niko.hellsten-at-stratum.fi GSM: +358-(0)440-955301
==============================Original Headers============================== 7, 22 -- From niko.hellsten-at-stratum.fi Fri Apr 20 04:50:21 2007 7, 22 -- Received: from emh03.mail.saunalahti.fi (emh03.mail.saunalahti.fi [62.142.5.109]) 7, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3K9oK1j006613 7, 22 -- for {Microscopy-at-Microscopy.com} ; Fri, 20 Apr 2007 04:50:20 -0500 7, 22 -- Received: from Evkan (unknown [195.255.124.215]) 7, 22 -- by emh03.mail.saunalahti.fi (Postfix) with SMTP id 341A5158C6C 7, 22 -- for {Microscopy-at-Microscopy.com} ; Fri, 20 Apr 2007 12:50:18 +0300 (EEST) 7, 22 -- Message-ID: {002f01c78331$57183970$d77cffc3-at-stratum.fi} 7, 22 -- From: "Niko Hellsten" {niko.hellsten-at-stratum.fi} 7, 22 -- To: "Microscopy" {Microscopy-at-Microscopy.com} 7, 22 -- Subject: cross section sample preparation 7, 22 -- Date: Fri, 20 Apr 2007 12:50:26 +0300 7, 22 -- MIME-Version: 1.0 7, 22 -- Content-Type: text/plain; 7, 22 -- format=flowed; 7, 22 -- charset="iso-8859-1"; 7, 22 -- reply-type=original 7, 22 -- Content-Transfer-Encoding: 8bit 7, 22 -- X-Priority: 3 7, 22 -- X-MSMail-Priority: Normal 7, 22 -- X-Mailer: Microsoft Outlook Express 6.00.2900.3028 7, 22 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.3028 ==============================End of - Headers==============================
==============================Original Headers============================== 28, 24 -- From Joseph_Oparowski-at-bose.com Fri Apr 20 06:31:21 2007 28, 24 -- Received: from BOSEMX02.bose.com (bosemx02.bose.com [139.68.146.21]) 28, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id l3KBVLtH030547 28, 24 -- for {Microscopy-at-Microscopy.Com} ; Fri, 20 Apr 2007 06:31:21 -0500 28, 24 -- Received: from USMAFREXMB02.bose.com ([139.68.132.238]) by USMAFREXCN02.bose.com with Microsoft SMTPSVC(6.0.3790.1830); 28, 24 -- Fri, 20 Apr 2007 07:31:20 -0400 28, 24 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 28, 24 -- Content-class: urn:content-classes:message 28, 24 -- MIME-Version: 1.0 28, 24 -- Content-Type: text/plain; 28, 24 -- charset="iso-8859-1" 28, 24 -- Subject: RE: [Microscopy] LM-cross section sample preparation 28, 24 -- Date: Fri, 20 Apr 2007 07:31:20 -0400 28, 24 -- Message-ID: {210C73AB4F6E0B4FBCB77B969C0BD84A05FB9B26-at-USMAFREXMB02.bose.com} 28, 24 -- In-Reply-To: {200704200955.l3K9tAXI016454-at-ns.microscopy.com} 28, 24 -- X-MS-Has-Attach: 28, 24 -- X-MS-TNEF-Correlator: 28, 24 -- Thread-Topic: [Microscopy] LM-cross section sample preparation 28, 24 -- Thread-Index: AceDMggrilIqG/IDTj60rZrj7f5kLwACogaA 28, 24 -- From: "Oparowski, Joseph" {Joseph_Oparowski-at-bose.com} 28, 24 -- To: {niko.hellsten-at-stratum.fi} , {Microscopy-at-Microscopy.Com} 28, 24 -- X-OriginalArrivalTime: 20 Apr 2007 11:31:20.0785 (UTC) FILETIME=[6B303C10:01C7833F] 28, 24 -- Content-Transfer-Encoding: 8bit 28, 24 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l3KBVLtH030547 ==============================End of - Headers==============================
our institution has acquired money to buy an ultramicrotome for TEM sample preparation of material samples, such as layered structures (clay minerals or thin layers on Si support).
To my knowledge, there are currently two ultramicrotomes on the market - EM UC6 from Leica and Power-Tome XL from RMC. I have talked to sales people from both companies, and of course they both claim that their cutting technique is the best for cutting hard materials because it does not introduce internal vibrations. RMC is motor driven while Leica boasts with "the Gravity Stroke". I can imagine both techniques having some troubles vibrationwise so I am torn and confused. So here comes the question. Has anybody ever tried to section hard materials on these machines and compared the results?
Jessica, have you attempted ICC following routine aldehyde fixation and osmium post-fixation followed by standard epoxy embedding? The osmium will preserve the membranes and you *may* still get successful ICC. Give a holler and I'll send you a (work in progress) protocol that appears to yield successful IHC on LM thick sections using RT reagents (no microwaves, no heated steam baths).
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
"The ultimate human freedom is the ability to choose one's attitude in a given set of circumstances." -Viktor Frankl
-----Original Message----- X-from: cervantes-at-bendres.com [mailto:cervantes-at-bendres.com] Sent: Thursday, April 19, 2007 7:03 PM To: Bobrowski, Walter
Did I miss the replies to this query somehow? Is there a technical problem, or were the replies private? Would it be possible to post them?
Let's not be shy (if that's it) about posting to the whole list. I've gotten so much out of this resource (especially lately). I've had to re-post replies to my query for others who have had the same question and hadn't gotten the messages.
Thanks for keeping the information flowing; it's invaluable. Jessica Cervantes Bend Research Inc Bend, OR
-----Original Message----- X-from: drk-at-SHCC.org [mailto:drk-at-SHCC.org] Sent: Wednesday, April 18, 2007 10:49 AM To: Cervantes, Jessica
Fellow Microscopists,
A few days ago I posted a query for suggestions on how to stabilize lysosomal membranes in cultured cells prior to immunocytochemistry. We have tried several of the tips but without success. Thanks to those who replied! We have been following the condition of the lysosomes using a GFP tag that we know to be specifically compartmentalized. We note that following fixation and a rinse in TRIS-Glycine the GFP is still compartmentalized. However, perhaps in 90% EtOH and definitely in 1:1 LR White:90% EtOH we see the GFP moves from the lysosomes to the cytoplasm and nucleus. It seems that ethanol and certainly LR White contribute to the deterioration of membranes. Can anyone suggest an embedding media that might be a little gentler on membranes and still possibly work for surface label immunocytochemistry? We are thinking of trying Nanoplast, a water soluble media but any suggestions are welcome.
Thanks! Doug
Douglas R. Keene Assistant Investigator Micro-Imaging Center Shriners Hospital for Children 3102 S.W. Sam Jackson Park Road Portland, Oregon 97239 503-221-3434 drk-at-shcc.org
==============================Original Headers============================== 7, 22 -- From drk-at-SHCC.org Wed Apr 18 12:43:18 2007 7, 22 -- Received: from mail.SHCC.org (mail.shcc.org [64.213.211.202]) 7, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3IHhITP002098 7, 22 -- for {Microscopy-at-Microscopy.Com} ; Wed, 18 Apr 2007 12:43:18 -0500 7, 22 -- Received: from DRK2 ([64.213.211.62]) by mail.SHCC.org (PMDF V6.3 #31473) 7, 22 -- with ESMTPA id {0JGP0029FFTXS8-at-mail.SHCC.org} for Microscopy-at-Microscopy.Com; 7, 22 -- Wed, 18 Apr 2007 10:41:57 -0700 (PDT) 7, 22 -- Date: Wed, 18 Apr 2007 10:43:58 -0700 7, 22 -- From: Doug Keene {drk-at-SHCC.org} 7, 22 -- Subject: stabilization ofmembranes 7, 22 -- Sender: drk-at-SHCC.org 7, 22 -- To: Microscopy-at-Microscopy.Com 7, 22 -- Cc: drk-at-SHCC.org 7, 22 -- Reply-to: drk-at-SHCC.org 7, 22 -- Message-id: {0JGP0029GFTXS8-at-mail.SHCC.org} 7, 22 -- Organization: Shriners Hospitals for Children 7, 22 -- MIME-version: 1.0 7, 22 -- X-MIMEOLE: Produced By Microsoft MimeOLE V6.00.2900.3028 7, 22 -- X-Mailer: Microsoft Office Outlook, Build 11.0.5510 7, 22 -- Content-type: text/plain; charset=us-ascii 7, 22 -- Content-transfer-encoding: 7BIT 7, 22 -- Thread-Index: AceB4SRWHFc2k9QuQCCG4KbQ3H8v4A== ==============================End of - Headers==============================
==============================Original Headers============================== 13, 16 -- From cervantes-at-bendres.com Thu Apr 19 17:57:57 2007 13, 16 -- Received: from BRIEX04A.bri.local (bc161112.bendcable.com [216.228.161.112]) 13, 16 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3JMvuoK002012 13, 16 -- for {Microscopy-at-microscopy.com} ; Thu, 19 Apr 2007 17:57:56 -0500 13, 16 -- MIME-Version: 1.0 13, 16 -- Content-Type: text/plain; 13, 16 -- charset="us-ascii" 13, 16 -- Subject: RE: [Microscopy] stabilization of membranes 13, 16 -- Date: Thu, 19 Apr 2007 15:57:55 -0700 13, 16 -- Message-ID: {8943D65F9AD70E4488AD6DE09F15088768C47F-at-BRIEX04A} 13, 16 -- In-Reply-To: {200704181748.l3IHmsrE012175-at-ns.microscopy.com} 13, 16 -- References: {200704181748.l3IHmsrE012175-at-ns.microscopy.com} 13, 16 -- From: "Cervantes, Jessica" {cervantes-at-bendres.com} 13, 16 -- To: {Microscopy-at-microscopy.com} 13, 16 -- Content-Transfer-Encoding: 8bit 13, 16 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l3JMvuoK002012 ==============================End of - Headers==============================
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==============================Original Headers============================== 27, 31 -- From Walter.Bobrowski-at-pfizer.com Fri Apr 20 07:16:23 2007 27, 31 -- Received: from gromsgom01.pfizer.com (gromsgo.pfizer.com [148.168.224.84]) 27, 31 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3KCGM21022819 27, 31 -- for {microscopy-at-microscopy.com} ; Fri, 20 Apr 2007 07:16:22 -0500 27, 31 -- Received: from mopamrexc01.amer.pfizer.com (mopamrexc01.pfizer.com [170.116.32.254]) 27, 31 -- by gromsgom01.pfizer.com (8.13.7/8.13.7) with ESMTP id l3KCGI65016247; 27, 31 -- Fri, 20 Apr 2007 08:16:20 -0400 27, 31 -- Received: from mopamrexc03.amer.pfizer.com ([170.116.30.69]) by mopamrexc01.amer.pfizer.com with Microsoft SMTPSVC(6.0.3790.1830); 27, 31 -- Fri, 20 Apr 2007 08:16:19 -0400 27, 31 -- Received: from anaamrexm01.amer.pfizer.com ([162.48.101.10]) by mopamrexc03.amer.pfizer.com with Microsoft SMTPSVC(6.0.3790.1830); 27, 31 -- Fri, 20 Apr 2007 08:16:19 -0400 27, 31 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 27, 31 -- Content-class: urn:content-classes:message 27, 31 -- MIME-Version: 1.0 27, 31 -- Content-Type: text/plain; 27, 31 -- charset="us-ascii" 27, 31 -- Subject: RE: [Microscopy] RE: stabilization of membranes 27, 31 -- Date: Fri, 20 Apr 2007 08:16:18 -0400 27, 31 -- Message-ID: {CF89AF6ABD93B746A55BCCD9480A4B3D08F62185-at-anaamrexm01.amer.pfizer.com} 27, 31 -- In-Reply-To: {200704192302.l3JN2u1n010853-at-ns.microscopy.com} 27, 31 -- X-MS-Has-Attach: 27, 31 -- X-MS-TNEF-Correlator: 27, 31 -- Thread-Topic: [Microscopy] RE: stabilization of membranes 27, 31 -- thread-index: AceC1t8j+2KQVeouTMe/hn6vqLmCnAAbdA8g 27, 31 -- From: "Bobrowski, Walter" {Walter.Bobrowski-at-pfizer.com} 27, 31 -- To: {cervantes-at-bendres.com} , {microscopy-at-microscopy.com} 27, 31 -- X-OriginalArrivalTime: 20 Apr 2007 12:16:19.0005 (UTC) FILETIME=[B373EED0:01C78345] 27, 31 -- X-Proofpoint-Virus-Version: vendor=fsecure engine=4.65.5502:2.3.11,1.2.37,4.0.164 definitions=2007-04-20_02:2007-04-19,2007-04-20,2007-04-20 signatures=0 27, 31 -- X-Proofpoint-Spam-Reason: safe 27, 31 -- Content-Transfer-Encoding: 8bit 27, 31 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l3KCGM21022819 ==============================End of - Headers==============================
To avoid smearing, especially in the final stages of polishing, try using Buehler's MasterMet and MasterPrep on a spongey pad such as their Chemocloth. This is good for flatness and preserving delicate features even with layers of very different materials.
Alan Stone ASTON
Note: Buehler is not the only supplier of these products. Other metallographic suppliers may have similar products, but I am not familiar with them.
At 04:51 AM 4/20/2007, you wrote:
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==============================Original Headers============================== 15, 24 -- From as-at-astonmet.com Fri Apr 20 07:42:48 2007 15, 24 -- Received: from outbound3.mail.tds.net (outbound3.mail.tds.net [216.170.230.93]) 15, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3KCgldd002379 15, 24 -- for {microscopy-at-microscopy.com} ; Fri, 20 Apr 2007 07:42:47 -0500 15, 24 -- Received: from outaamta02.mail.tds.net (outaamta02.mail.tds.net [216.170.230.32]) 15, 24 -- by outbound3.mail.tds.net (8.13.6/8.13.4) with ESMTP id l3KCgktl001817; 15, 24 -- Fri, 20 Apr 2007 07:42:47 -0500 15, 24 -- Received: from Alan.astonmet.com ([69.11.219.4]) by outaamta02.mail.tds.net 15, 24 -- with ESMTP 15, 24 -- id {20070420124246.KPFE21812.outaamta02.mail.tds.net-at-Alan.astonmet.com} ; 15, 24 -- Fri, 20 Apr 2007 07:42:46 -0500 15, 24 -- X-Mailer: QUALCOMM Windows Eudora Version 7.1.0.9 15, 24 -- Date: Fri, 20 Apr 2007 07:42:53 -0500 15, 24 -- To: microscopy-at-microscopy.com 15, 24 -- From: Alan Stone {as-at-astonmet.com} 15, 24 -- Subject: Re: [Microscopy] cross section sample preparation 15, 24 -- Cc: niko.hellsten-at-stratum.fi 15, 24 -- In-Reply-To: {200704200951.l3K9pZRO008693-at-ns.microscopy.com} 15, 24 -- References: {200704200951.l3K9pZRO008693-at-ns.microscopy.com} 15, 24 -- Mime-Version: 1.0 15, 24 -- Content-Type: text/plain; charset="iso-8859-1"; format=flowed 15, 24 -- Message-Id: {20070420124246.KPFE21812.outaamta02.mail.tds.net-at-Alan.astonmet.com} 15, 24 -- Content-Transfer-Encoding: 8bit 15, 24 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l3KCgldd002379 ==============================End of - Headers==============================
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Title-Subject: [Filtered] Stereology : grids and methods
Question: To anyone who could be give me a tip...
We are investigating grid counting for particles counting in feed, but also taking into account their relative size : a particle being 5 times bigger than all others should be counted as 5 instead of 1). Basically we intend to use eyepiece square grids reticles. Does anyone has other proposals (eg type of grid) ? We are also looking for a good reference method description including a discussion on biases (such as that of counting only two adjacent lengths of the square...), is there an ultimate good and recent book of reference available ?
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Email: fab-at-tariffenet.it Name: fabio
Organization: University of Catania
Title-Subject: [Filtered] need help fo digital camera
Question: Dear All, I would like to mount a digital camera (such as a Canon Powershot A-540 or A-560)on my Reichert Microstar IV light microscope (equipped with a trinocular viewing body). I know I need an adaper for the camera (LADC52F) and an adapter for the microscope trinocular body. Which kind of adapter I need? Any suggestions?
Thank You
Fabio University of Catania, Italy. fab-at-tariffenet.it
Looks like 2 badly collapsed mite larvae. Given the scale bar on the lower-mag shot, these are probably recently hatched. The long, fuzzy things sticking out of the circles are sensory setae. These are dorsal views, and the legs are the long, straight fuzzy things that some out from underneath them. The confusing bunch of things sticking out of the end of the one larva (whose posterior end is under the 2nd larva) are the pedipalps and chelicerae (soft parts at this stage). The 2 larger structures sticking "north" out of the mess are the forelegs, and joints are just visible. No clue about the taxon, but judging by size, I'd guess Astigmata, or whatever that group is these days.
Mind, this could all be empty arm-waving, but I'd be willing to bet a pitcher of good beer on it.
Phil
} Hello All, } } Do any of you know what this thing is? It looks like a "bug" but it's } attached to a probable piece of fungus. Any ideas welcome, this is a head } scratcher for everyone that has seen it. } } Thanks! } } Tom Kaye } } Tomkaye.com/images/fungus_lrg.jpg Long shot, the bug thing is lower right } } } Tomkaye.com/images/fungus2_lrg.jpg Close up. -- 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 Apr 20 08:02:39 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 l3KD2dZs004819 5, 22 -- for {Microscopy-at-microscopy.com} ; Fri, 20 Apr 2007 08:02:39 -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 l3KDQq6l023538 5, 22 -- for {Microscopy-at-microscopy.com} ; Fri, 20 Apr 2007 09:26:57 -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, 20 Apr 2007 09:02:38 -0400 5, 22 -- Mime-Version: 1.0 5, 22 -- Message-Id: {f06230903c24e6872a0f3-at-[141.209.160.249]} 5, 22 -- In-Reply-To: {200704200010.l3K0Akaf000519-at-ns.microscopy.com} 5, 22 -- References: {200704200010.l3K0Akaf000519-at-ns.microscopy.com} 5, 22 -- Date: Fri, 20 Apr 2007 09:02:36 -0400 5, 22 -- To: Microscopy-at-microscopy.com 5, 22 -- From: Philip Oshel {oshel1pe-at-cmich.edu} 5, 22 -- Subject: Re: [Microscopy] Need help with ID 5, 22 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 5, 22 -- X-OriginalArrivalTime: 20 Apr 2007 13:02:38.0161 (UTC) FILETIME=[2BF57C10:01C7834C] 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==============================
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Email: j.janssen-at-nki.nl Name: J.W.R.M. Janssen
Organization: Dutch Cancer Institute
Title-Subject: [Filtered] grids and methods
Question: Dear Pascal,
Here is a good reference: Recent developments for quantifying immunogold label on transmission EM thin sections Terry M Mayhew Centre for Integrated Systems Biology & Medicine, School of Biomedical Sciences, University of Nottingham, UK Succes, Hans.
I am a relative newcomer to the microscopy/histology field and need some advice on sample preparation. I am trying to section a multilayer packaging film (e.g. polyester, aluminum foil, polyethylene lamination) and am having trouble keeping the sample flat, so I can get a good reading on individual layer thicknesses. I am using a cryostat microtome for sample preparation. Is there a simple technique I can employ, or a type of embedding compound that works better for industrial applications? Thanks to any who can give me a hand, I appreciate it.
R. Jefferson Babbitt, Ph.D. Analytical Services Manager Fres-co System USA 3005 State Rd. Telford, PA 18969-1021 voice - 215-721-4600 x2149 cell - 267-236-4027 fax - 215-799-8017 email - jbabbitt-at-fresco.com
==============================Original Headers============================== 2, 24 -- From JBABBITT-at-fresco.com Fri Apr 20 10:44:43 2007 2, 24 -- Received: from citrix.fresco.local (mail.fresco.com [12.104.46.34]) 2, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id l3KFihRr031948 2, 24 -- for {Microscopy-at-Microscopy.Com} ; Fri, 20 Apr 2007 10:44:43 -0500 2, 24 -- Received: from Exchange.fresco.local ([137.1.3.24]) 2, 24 -- by citrix.fresco.local (SMSSMTP 4.1.14.46) with SMTP id M2007042011475624848 2, 24 -- for {Microscopy-at-Microscopy.Com} ; Fri, 20 Apr 2007 11:47:56 -0400 2, 24 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 2, 24 -- Content-class: urn:content-classes:message 2, 24 -- MIME-Version: 1.0 2, 24 -- Content-Type: text/plain; 2, 24 -- charset="iso-8859-1" 2, 24 -- Subject: RE: Cross-sectioning packaging film 2, 24 -- Date: Fri, 20 Apr 2007 11:44:42 -0400 2, 24 -- Message-ID: {2BA2AD943995314D9D98D68F33A0AC7B035B7776-at-exchange.fresco.local} 2, 24 -- In-Reply-To: {2BA2AD943995314D9D98D68F33A0AC7B035B7774-at-exchange.fresco.local} 2, 24 -- X-MS-Has-Attach: 2, 24 -- X-MS-TNEF-Correlator: 2, 24 -- Thread-Topic: Cross-sectioning packaging film 2, 24 -- Thread-Index: AceDUI+XLo+FTSxXT9atWsIzk0rybQAEiOBA 2, 24 -- From: "Babbitt, Jeff" {JBABBITT-at-fresco.com} 2, 24 -- To: {Microscopy-at-Microscopy.Com} 2, 24 -- Content-Transfer-Encoding: 8bit 2, 24 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l3KFihRr031948 ==============================End of - Headers==============================
Thanks Walter. I'm definitely interested in your protocol.
Jessica Cervantes Bend Research Inc Bend, OR
-----Original Message----- X-from: Walter.Bobrowski-at-pfizer.com [mailto:Walter.Bobrowski-at-pfizer.com] Sent: Friday, April 20, 2007 5:22 AM To: Cervantes, Jessica
Jessica, have you attempted ICC following routine aldehyde fixation and osmium post-fixation followed by standard epoxy embedding? The osmium will preserve the membranes and you *may* still get successful ICC. Give a holler and I'll send you a (work in progress) protocol that appears to yield successful IHC on LM thick sections using RT reagents (no microwaves, no heated steam baths).
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
"The ultimate human freedom is the ability to choose one's attitude in a given set of circumstances." -Viktor Frankl
-----Original Message----- X-from: cervantes-at-bendres.com [mailto:cervantes-at-bendres.com] Sent: Thursday, April 19, 2007 7:03 PM To: Bobrowski, Walter
Did I miss the replies to this query somehow? Is there a technical problem, or were the replies private? Would it be possible to post them?
Let's not be shy (if that's it) about posting to the whole list. I've gotten so much out of this resource (especially lately). I've had to re-post replies to my query for others who have had the same question and hadn't gotten the messages.
Thanks for keeping the information flowing; it's invaluable. Jessica Cervantes Bend Research Inc Bend, OR
-----Original Message----- X-from: drk-at-SHCC.org [mailto:drk-at-SHCC.org] Sent: Wednesday, April 18, 2007 10:49 AM To: Cervantes, Jessica
Fellow Microscopists,
A few days ago I posted a query for suggestions on how to stabilize lysosomal membranes in cultured cells prior to immunocytochemistry. We have tried several of the tips but without success. Thanks to those who replied! We have been following the condition of the lysosomes using a GFP tag that we know to be specifically compartmentalized. We note that following fixation and a rinse in TRIS-Glycine the GFP is still compartmentalized. However, perhaps in 90% EtOH and definitely in 1:1 LR White:90% EtOH we see the GFP moves from the lysosomes to the cytoplasm and nucleus. It seems that ethanol and certainly LR White contribute to the deterioration of membranes. Can anyone suggest an embedding media that might be a little gentler on membranes and still possibly work for surface label immunocytochemistry? We are thinking of trying Nanoplast, a water soluble media but any suggestions are welcome.
Thanks! Doug
Douglas R. Keene Assistant Investigator Micro-Imaging Center Shriners Hospital for Children 3102 S.W. Sam Jackson Park Road Portland, Oregon 97239 503-221-3434 drk-at-shcc.org
==============================Original Headers============================== 7, 22 -- From drk-at-SHCC.org Wed Apr 18 12:43:18 2007 7, 22 -- Received: from mail.SHCC.org (mail.shcc.org [64.213.211.202]) 7, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3IHhITP002098 7, 22 -- for {Microscopy-at-Microscopy.Com} ; Wed, 18 Apr 2007 12:43:18 -0500 7, 22 -- Received: from DRK2 ([64.213.211.62]) by mail.SHCC.org (PMDF V6.3 #31473) 7, 22 -- with ESMTPA id {0JGP0029FFTXS8-at-mail.SHCC.org} for Microscopy-at-Microscopy.Com; 7, 22 -- Wed, 18 Apr 2007 10:41:57 -0700 (PDT) 7, 22 -- Date: Wed, 18 Apr 2007 10:43:58 -0700 7, 22 -- From: Doug Keene {drk-at-SHCC.org} 7, 22 -- Subject: stabilization ofmembranes 7, 22 -- Sender: drk-at-SHCC.org 7, 22 -- To: Microscopy-at-Microscopy.Com 7, 22 -- Cc: drk-at-SHCC.org 7, 22 -- Reply-to: drk-at-SHCC.org 7, 22 -- Message-id: {0JGP0029GFTXS8-at-mail.SHCC.org} 7, 22 -- Organization: Shriners Hospitals for Children 7, 22 -- MIME-version: 1.0 7, 22 -- X-MIMEOLE: Produced By Microsoft MimeOLE V6.00.2900.3028 7, 22 -- X-Mailer: Microsoft Office Outlook, Build 11.0.5510 7, 22 -- Content-type: text/plain; charset=us-ascii 7, 22 -- Content-transfer-encoding: 7BIT 7, 22 -- Thread-Index: AceB4SRWHFc2k9QuQCCG4KbQ3H8v4A== ==============================End of - Headers==============================
==============================Original Headers============================== 13, 16 -- From cervantes-at-bendres.com Thu Apr 19 17:57:57 2007 13, 16 -- Received: from BRIEX04A.bri.local (bc161112.bendcable.com [216.228.161.112]) 13, 16 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3JMvuoK002012 13, 16 -- for {Microscopy-at-microscopy.com} ; Thu, 19 Apr 2007 17:57:56 -0500 13, 16 -- MIME-Version: 1.0 13, 16 -- Content-Type: text/plain; 13, 16 -- charset="us-ascii" 13, 16 -- Subject: RE: [Microscopy] stabilization of membranes 13, 16 -- Date: Thu, 19 Apr 2007 15:57:55 -0700 13, 16 -- Message-ID: {8943D65F9AD70E4488AD6DE09F15088768C47F-at-BRIEX04A} 13, 16 -- In-Reply-To: {200704181748.l3IHmsrE012175-at-ns.microscopy.com} 13, 16 -- References: {200704181748.l3IHmsrE012175-at-ns.microscopy.com} 13, 16 -- From: "Cervantes, Jessica" {cervantes-at-bendres.com} 13, 16 -- To: {Microscopy-at-microscopy.com} 13, 16 -- Content-Transfer-Encoding: 8bit 13, 16 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l3JMvuoK002012 ==============================End of - Headers==============================
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==============================Original Headers============================== 27, 31 -- From Walter.Bobrowski-at-pfizer.com Fri Apr 20 07:16:23 2007 27, 31 -- Received: from gromsgom01.pfizer.com (gromsgo.pfizer.com [148.168.224.84]) 27, 31 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3KCGM21022819 27, 31 -- for {microscopy-at-microscopy.com} ; Fri, 20 Apr 2007 07:16:22 -0500 27, 31 -- Received: from mopamrexc01.amer.pfizer.com (mopamrexc01.pfizer.com [170.116.32.254]) 27, 31 -- by gromsgom01.pfizer.com (8.13.7/8.13.7) with ESMTP id l3KCGI65016247; 27, 31 -- Fri, 20 Apr 2007 08:16:20 -0400 27, 31 -- Received: from mopamrexc03.amer.pfizer.com ([170.116.30.69]) by mopamrexc01.amer.pfizer.com with Microsoft SMTPSVC(6.0.3790.1830); 27, 31 -- Fri, 20 Apr 2007 08:16:19 -0400 27, 31 -- Received: from anaamrexm01.amer.pfizer.com ([162.48.101.10]) by mopamrexc03.amer.pfizer.com with Microsoft SMTPSVC(6.0.3790.1830); 27, 31 -- Fri, 20 Apr 2007 08:16:19 -0400 27, 31 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 27, 31 -- Content-class: urn:content-classes:message 27, 31 -- MIME-Version: 1.0 27, 31 -- Content-Type: text/plain; 27, 31 -- charset="us-ascii" 27, 31 -- Subject: RE: [Microscopy] RE: stabilization of membranes 27, 31 -- Date: Fri, 20 Apr 2007 08:16:18 -0400 27, 31 -- Message-ID: {CF89AF6ABD93B746A55BCCD9480A4B3D08F62185-at-anaamrexm01.amer.pfizer.com} 27, 31 -- In-Reply-To: {200704192302.l3JN2u1n010853-at-ns.microscopy.com} 27, 31 -- X-MS-Has-Attach: 27, 31 -- X-MS-TNEF-Correlator: 27, 31 -- Thread-Topic: [Microscopy] RE: stabilization of membranes 27, 31 -- thread-index: AceC1t8j+2KQVeouTMe/hn6vqLmCnAAbdA8g 27, 31 -- From: "Bobrowski, Walter" {Walter.Bobrowski-at-pfizer.com} 27, 31 -- To: {cervantes-at-bendres.com} , {microscopy-at-microscopy.com} 27, 31 -- X-OriginalArrivalTime: 20 Apr 2007 12:16:19.0005 (UTC) FILETIME=[B373EED0:01C78345] 27, 31 -- X-Proofpoint-Virus-Version: vendor=fsecure engine=4.65.5502:2.3.11,1.2.37,4.0.164 definitions=2007-04-20_02:2007-04-19,2007-04-20,2007-04-20 signatures=0 27, 31 -- X-Proofpoint-Spam-Reason: safe 27, 31 -- Content-Transfer-Encoding: 8bit 27, 31 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l3KCGM21022819 ==============================End of - Headers==============================
==============================Original Headers============================== 32, 17 -- From cervantes-at-bendres.com Fri Apr 20 10:47:39 2007 32, 17 -- Received: from BRIEX04A.bri.local (bc161112.bendcable.com [216.228.161.112]) 32, 17 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3KFlban003778 32, 17 -- for {Microscopy-at-microscopy.com} ; Fri, 20 Apr 2007 10:47:39 -0500 32, 17 -- MIME-Version: 1.0 32, 17 -- Content-Type: text/plain; 32, 17 -- charset="us-ascii" 32, 17 -- Subject: RE: [Microscopy] stabilization of membranes 32, 17 -- Date: Fri, 20 Apr 2007 08:47:36 -0700 32, 17 -- Message-ID: {8943D65F9AD70E4488AD6DE09F15088768C481-at-BRIEX04A} 32, 17 -- In-Reply-To: {200704201222.l3KCMO5u032747-at-ns.microscopy.com} 32, 17 -- References: {200704201222.l3KCMO5u032747-at-ns.microscopy.com} 32, 17 -- From: "Cervantes, Jessica" {cervantes-at-bendres.com} 32, 17 -- To: {Walter.Bobrowski-at-pfizer.com} 32, 17 -- Cc: {Microscopy-at-microscopy.com} 32, 17 -- Content-Transfer-Encoding: 8bit 32, 17 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l3KFlban003778 ==============================End of - Headers==============================
I can see your problem. You have a soft material that smears with relatively hard materials and they have much different chemistries. I would like to suggest an alternative to bringing out your structure. After polishing, instead of using a chemical etch, you can use ion etching. What I would do is to ion polish at a low angle for about 10 minutes and then follow that with a high angle etch at about 35 degrees for a short time. With copper samples for light microscopy, we found that 2.5 min gave a good grain appearance at 500X and could see the interfaces with elelctroless copper layers. Since the ion polishing and etching is a physical process, it is less sensitive to the chemical nature of the materials in your cross section. We make and sell the IBS/e that can be used for this purpose and have gotten very nice results with electroplated copper samples with light microscopy. Gatan has a nicely prepared booklet that shows light microscopy results as well.
Please contact me offline and we could arrange to run some samples for you. I would only ask that we could put the results on our web site as an application note.
Disclaimer: South Bay Techonology, Inc. makes and sells the IBS/e ion beams sputter and etch system as well as metallurgical polishing equipment.
-Scott
Scott D. Walck, Ph.D. Technical Director South Bay Technology, Inc. 1120 Via Callejon San Clemente, CA 92673
-------- Original Message -------- } From: niko.hellsten-at-stratum.fi } Sent: Friday, April 20, 2007 5:54 AM } To: Walck-at-SouthBayTech.com } Subject: [Microscopy] 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 } ---------------------------------------------------------------------------- } } Hello everyone } } I'm doing basic cross section preparation for light microscopy work. My } samples are stainless steel plated with 0-50 micrometers thick layers of } copper and chromium. } } I would like to make the interfaces clearer than they are after } grinding/polishing. I have thought about etching/corroding the surface with } some acid or other chemical. } } I would like to hear from the experts what kind of chemicals they might } recommend. Or if etching my samples chemically would do me any good at all. } } Thanks in advance } } Niko Hellstén } Product Engineer } Stratum Oy } mail: niko.hellsten-at-stratum.fi } GSM: +358-(0)440-955301 } } } ==============================Original Headers============================== } 7, 22 -- From niko.hellsten-at-stratum.fi Fri Apr 20 04:50:21 2007 } 7, 22 -- Received: from emh03.mail.saunalahti.fi (emh03.mail.saunalahti.fi [62.142.5.109]) } 7, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3K9oK1j006613 } 7, 22 -- for {Microscopy-at-Microscopy.com} ; Fri, 20 Apr 2007 04:50:20 -0500 } 7, 22 -- Received: from Evkan (unknown [195.255.124.215]) } 7, 22 -- by emh03.mail.saunalahti.fi (Postfix) with SMTP id 341A5158C6C } 7, 22 -- for {Microscopy-at-Microscopy.com} ; Fri, 20 Apr 2007 12:50:18 +0300 (EEST) } 7, 22 -- Message-ID: {002f01c78331$57183970$d77cffc3-at-stratum.fi} } 7, 22 -- From: "Niko Hellsten" {niko.hellsten-at-stratum.fi} } 7, 22 -- To: "Microscopy" {Microscopy-at-Microscopy.com} } 7, 22 -- Subject: cross section sample preparation } 7, 22 -- Date: Fri, 20 Apr 2007 12:50:26 +0300 } 7, 22 -- MIME-Version: 1.0 } 7, 22 -- Content-Type: text/plain; } 7, 22 -- format=flowed; } 7, 22 -- charset="iso-8859-1"; } 7, 22 -- reply-type=original } 7, 22 -- Content-Transfer-Encoding: 8bit } 7, 22 -- X-Priority: 3 } 7, 22 -- X-MSMail-Priority: Normal } 7, 22 -- X-Mailer: Microsoft Outlook Express 6.00.2900.3028 } 7, 22 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.3028 } ==============================End of - Headers==============================
==============================Original Headers============================== 5, 20 -- From walck-at-southbaytech.com Fri Apr 20 11:18:38 2007 5, 20 -- Received: from smtp15.dc2.safesecureweb.com (smtp15.dc2.safesecureweb.com [65.36.255.249]) 5, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3KGIcXU023227 5, 20 -- for {microscopy-at-microscopy.com} ; Fri, 20 Apr 2007 11:18:38 -0500 5, 20 -- Received: from mail43.safesecureweb.com (barrage.ad.safesecureweb.com [10.10.1.193]) 5, 20 -- by smtp15.dc2.safesecureweb.com (Spam Firewall) with ESMTP 5, 20 -- id D543E1778CB7; Fri, 20 Apr 2007 12:18:37 -0400 (EDT) 5, 20 -- MIME-Version: 1.0 5, 20 -- Date: Fri, 20 Apr 2007 12:17:56 -0400 5, 20 -- Received: from [72.197.40.68] by mail43.safesecureweb.com via HTTP; Fri, 20 Apr 2007 12:17:56 -0400 5, 20 -- Content-Type: text/plain; 5, 20 -- charset=iso-8859-1 5, 20 -- Subject: re: [Microscopy] cross section sample preparation 5, 20 -- From: "Scott Walck" {walck-at-southbaytech.com} 5, 20 -- Reply-To: Walck-at-southbaytech.com 5, 20 -- To: {niko.hellsten-at-stratum.fi} 5, 20 -- CC: {Microscopy-at-microscopy.com} 5, 20 -- Message-ID: {739462ad9e61403e9be222e48e7a08cd-at-southbaytech.com} 5, 20 -- Content-Transfer-Encoding: 8bit 5, 20 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l3KGIcXU023227 ==============================End of - Headers==============================
I would try freezing the sample and using a single bend fracture procedure as set out below. We have used this technique on polythene freezer bags and many other materials for SEM but the technique is equally applicable to LM.
"Over the many years that clients and I have been investigating the cross sections of materials by for the best method is to fracture the material. The SEM is very clever in that it sees a cut surface and tells us "this is a cross section cut with a sharp scalpel blade" or "this is a cross section cut with a blunt scalpel blade" etc etc. Preparation method A
1. Cut down the material to 1cm by 3cms place it into liquid nitrogen until it stops bubbling.
2. Remove the material and crack it using either heavy duty tweezers or fine pliers. If you are unable to crack the material and are forced to flex it in order for it to crack this is not good enough! In the latter case reduce or neck the material as shown, even a 0.5mm long crack could provide a great deal of detail in the SEM?
3. When the pieces have dried out (condensation) they may both be observed by LM and SEM
Fibres that will not fracture by the above method could be fractured by one of two other methods.
Method B
1. Insert the material in a small diameter tube (thin drinking straws are ideal). Cut the straw down to about 3cms tall. Block one end with wax, modelling clay or similar material.
2. Using a syringe force water into the straw and block the end as above.
3. Drop the straw into liquid nitrogen then follow method A part 2 above.
4. When the pieces have dried out (condensation) they may be observed by both LM and SEM
Method C
1. Drill 2mm to 3mm holes in a pair of stubs as shown in diagram 2.
2. Infiltrate the holes with a water soluble carbon solution and push a bundle of fibres through the carbon solution.
3. When dry follow method A part 2 except use a blade to initiate the crack
4. When the pieces have dried out (condensation) they may be observed by both LM and SEM
Method C has been used with materials like freezer bags that were failing. In this case the material was spun into a small spiral and then infiltrated with the carbon solution."
The above data was taken from our "hints and tips" web page.
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: {JBABBITT-at-fresco.com} To: {protrain-at-emcourses.com} Sent: Friday, April 20, 2007 4:45 PM
R. Jefferson Babbitt, Ph.D wrote the following: ================================================================ I am a relative newcomer to the microscopy/histology field and need some advice on sample preparation. I am trying to section a multilayer packaging film (e.g. polyester, aluminum foil, polyethylene lamination) and am having trouble keeping the sample flat, so I can get a good reading on individual layer thicknesses. I am using a cryostat microtome for sample preparation. Is there a simple technique I can employ, or a type of embedding compound that works better for industrial applications? Thanks to any who can give me a hand, I appreciate it. ======================================================================== We have done this same kind of system in our own laboratory. The quick-and-dirty way to get a cross-section is liquid nitrogen fracturing but as you point out, getting a good edge-on view is not necessarily easy. Another danger: If you don't know in advance exactly how many layers there are, one or more outer layers can split off further down from the fracture surface and what you think is the fracture surface indeed might not be the facture surface.
Hence, for SEM work, we always gold coat the two sides so that so long as we can see in the fracture surface the two gold layers, we know we have the entire cross-section, and in order to keep it "straight", we mount is using an angled SEM mount (available from SPI Supplies as well as all of our major competitors), which is then tilted 45 deg. for the head on view.
But we have pretty consistently found that a TEM view, while the sample work up is more tedious, is far more rewarding. Most of the time one wants to look at such films, is because there is an adhesion failure between one or more of the layers and the TEM view is able to resolve features (such as contaminants) between the layers or the dispersion of inorganic fillers in one or more of the layers. We still gold coat such films (with passivation layers) before embedding to ensure that there is not interaction of the embedding resin with the polymer film layers. Our preferred embedding resin is our own SPI-Pon 812 epoxy- based embedding resin (but we believe equivalent results will be obtained with the equivalent sold by others).
One other thing: The cross-sectioning, irrespective of what method you are using, must be done cryo, with an ultramicrotome using a diamond knife. A glass knife will not last very long against the aluminum foil layer.
Since you are a "neighbor" to SPI Supplies, we would be happy to have you visit and we would show you how we do these kinds of films.
Disclaimers: Mentioned in this posting are some of our own products and we have the vested interest in promoting their use.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
==============================Original Headers============================== 12, 35 -- From cgarber-at-2spi.com Fri Apr 20 12:20:50 2007 12, 35 -- Received: from relay3.scalera.ch (relay3.scalera.ch [195.129.94.189]) 12, 35 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id l3KHKnDb014966 12, 35 -- for {microscopy-at-microscopy.com} ; Fri, 20 Apr 2007 12:20:49 -0500 12, 35 -- Received: (qmail 10022 invoked by uid 89); 20 Apr 2007 17:20:47 -0000 12, 35 -- Received: from unknown (HELO blade1-4.iptech.localdomain) (195.129.94.130) 12, 35 -- by relay3.scalera.ch with SMTP; 20 Apr 2007 17:20:47 -0000 12, 35 -- Received: (qmail 22537 invoked by uid 104); 20 Apr 2007 17:20:47 -0000 12, 35 -- Received: from 193.247.86.162 by blade1-4 (envelope-from {cgarber-at-2spi.com} , uid 408) with qmail-scanner-1.25 12, 35 -- (clamdscan: 0.88.5. spamassassin: 3.1.5 12, 35 -- Clear:RC:0(193.247.86.162):SA:0(-1.4/5.0):. 12, 35 -- Processed in 0.598572 secs); 20 Apr 2007 17:20:47 -0000 12, 35 -- X-Spam-Checker-Version: SpamAssassin 3.1.8 (2007-02-13) on 12, 35 -- blade1-4.iptech.localdomain 12, 35 -- X-Spam-Level: 12, 35 -- X-Spam-Status: No, score=-1.4 required=5.0 tests=ALL_TRUSTED 12, 35 -- autolearn=disabled version=3.1.8 12, 35 -- X-Envelope-From: cgarber-at-2spi.com 12, 35 -- Received: from unknown (HELO ibm9x18xhqdz1o) (smtp-ithospitality-at-cablepower.ch-at-193.247.86.162) 12, 35 -- by blade1-4.iptech.localdomain with SMTP; 20 Apr 2007 17:20:46 -0000 12, 35 -- Message-ID: {001901c7836f$d69b02c0$3700000a-at-ibm9x18xhqdz1o} 12, 35 -- From: "Garber, Charles A." {cgarber-at-2spi.com} 12, 35 -- To: {microscopy-at-microscopy.com} 12, 35 -- Subject: Preparation of multilayer packaging films 12, 35 -- Date: Fri, 20 Apr 2007 13:17:54 -0400 12, 35 -- MIME-Version: 1.0 12, 35 -- Content-Type: text/plain; 12, 35 -- format=flowed; 12, 35 -- charset="iso-8859-1"; 12, 35 -- reply-type=original 12, 35 -- Content-Transfer-Encoding: 7bit 12, 35 -- X-Priority: 3 12, 35 -- X-MSMail-Priority: Normal 12, 35 -- X-Mailer: Microsoft Outlook Express 6.00.2900.3028 12, 35 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.3028 ==============================End of - Headers==============================
Many thanks to those who responded to my request for suggestions for stabilizing membranes (lysosomes in particular). My specific aim in this experiment is to stabilize cultured cells containing GFP tagged lysosomes for surface-label immunocytochemistry with the hope of co-localizing GFP expressing protein (using confocal images overlaid onto TEM images cut from the next serial section) and another potentially interactive protein localized with immunogold on the TEM section. We are able to follow GFP emission through the protocol using our confocal microscope. In cultures fixed in media buffered 4% paraformaldehyde/1% glutaraldehyde, we see that GFP remains compartmentalized up to 90% ethanol but after the introduction of 1:1 90% EtOH:LRWhite, GFP is no longer compartmentalized and instead is distributed throughout the cytoplasm and nucleus. I am now in the process of trying some of the ideas. A preliminary result suggests that the lysosomes are stabilized somewhat more by the inclusion of 4% picric acid; also by progressively lowering temperature during graded ethanol dehydration to minus 20C, embedding in Lowicryl HM20 at -20C, and polymerization at -20C. We have yet to section the HM20, but we hope that confocal microscopy on 0.5um sectioned HM20 will reveal compartmentalized GFP. Then we'll see what mess we've made of the additional antibody-binding epitopes. We love a challenge!
To address the request that the responses be shared, here is a summary. Since some of these responses were made privately, I have not included the author's names.
Response #1:
what may help: high-pressure immobilization (cryo-fixation; expensive but really good), followed by freeze-substitution at -90 (8 to 48 hrs)/-60 (8hrs)/-30 (6 to 8 hrs) as described in several papers.
It is worth trying: Aceton + 0.1/0.2% OsO4, +0.5% Uac +/-GA (0.1 to 1%) +/-FA (0.25 to 2%) (MANY variants possible, I know) or EtOH, no OsO4, but add some or all other chemicals/fixatives MeOH, no OsO4, but add some or all ... Aceton + 2%GA (ready available as it is) Aceton + 0.2%GA + UAc 0.5% ... and so on.
The advantage of cryo-substitution at very low temperatures: The chemicals are not reactive at low temperature initially (only slowly at -60, and then at higher temperatures). The chemicals become evenly distributed in the cell/tissue, and then react at the same time upon warming, everywhere, similarly. Result: tissue, cells and membranes are often better preserved. in addition, you may add some water (1 - 5%?) to the freeze-substitution medium, (YES!!), as suggested by Paul Walther a few years ago. We have done this, others as well, with success.
Response #2:
Have you tried tannic acid! It is supposed to stabilize plasma membranes, don't know about Lysosomal membranes,
Response #3"
While it would involve doing freeze-substitution, Lowicryl HM 20 is great for membranes. You would need to do low temperature dehydration after room temperature or 4 degree C aldehyde fixation. I used to use a small amount of glutaraldehyde with the methanol free buffered formalin. I also used uranyl acetate to help preserve membranes and dehydrated cold with methanol. You can look at my paper for details:
The Journal of Histochemistry and Cytochemistry 40(10):1491-1500, 1992 "Immunocytochemical Localization of Lysozyme and Surfactant Protein A in Rat Type II Cells and Extracellular Surfactant"
Response #4:
I have a couple of thoughts on the problem. I don't think you are getting any buffering capacity from the culture medium. And if there is any serum in the medium, you have titrated all the aldehydes with the serum and have no functional fixative left when you are trying to fix the cells.
Summary of additional responses:
These included longer fixation times, fixation at ambient temperature not 4C, and the importance of including Calcium in the fixation buffer.
Again, thank you to those who responded and to all interested microscopists!
Doug Keene Douglas R. Keene Assistant Investigator Micro-Imaging Center Shriners Hospital for Children 3102 S.W. Sam Jackson Park Road Portland, Oregon 97239 503-221-3434 drk-at-shcc.org
==============================Original Headers============================== 19, 22 -- From drk-at-SHCC.org Fri Apr 20 12:36:52 2007 19, 22 -- Received: from mail.SHCC.org (mail.shcc.org [64.213.211.202]) 19, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3KHap1k026613 19, 22 -- for {Microscopy-at-Microscopy.Com} ; Fri, 20 Apr 2007 12:36:52 -0500 19, 22 -- Received: from DRK2 ([64.213.211.62]) by mail.SHCC.org (PMDF V6.3 #31473) 19, 22 -- with ESMTPA id {0JGT008G14V30T-at-mail.SHCC.org} for Microscopy-at-Microscopy.Com; 19, 22 -- Fri, 20 Apr 2007 10:35:27 -0700 (PDT) 19, 22 -- Date: Fri, 20 Apr 2007 10:37:27 -0700 19, 22 -- From: Doug Keene {drk-at-SHCC.org} 19, 22 -- Subject: responses to membrane stabilization querry (long) 19, 22 -- Sender: drk-at-SHCC.org 19, 22 -- To: Microscopy-at-Microscopy.Com 19, 22 -- Cc: drk-at-SHCC.org 19, 22 -- Reply-to: drk-at-SHCC.org 19, 22 -- Message-id: {0JGT008G24V30T-at-mail.SHCC.org} 19, 22 -- Organization: Shriners Hospitals for Children 19, 22 -- MIME-version: 1.0 19, 22 -- X-MIMEOLE: Produced By Microsoft MimeOLE V6.00.2900.3028 19, 22 -- X-Mailer: Microsoft Office Outlook, Build 11.0.5510 19, 22 -- Content-type: text/plain; charset=us-ascii 19, 22 -- Content-transfer-encoding: 7BIT 19, 22 -- Thread-Index: AceDcpAds34GRAJXQ/ugUZSIbT2WoQ== ==============================End of - Headers==============================
Our group wants to buy a new SEM to replace an old AMRAY machine for failure analysis and quality control investigations. We also need EDS capabilities, and do not anticipate the need for any other analysis tools. It seems like we will need to go to 10 000X only or thereabouts, and we would like to have low vacuum capabilities for imaging nonmetallic components / painted parts. We want a large chamber, and a large range of stage motion (100 mm X 80 mm X 80 mm min.) We need the new machine to last a really long time, be easily maintained, and spare parts should be available for the indefinite future, as I do not anticipate funding for anything like this again for over a decade. I think that analog imaging is preferred to digital acquisition because of alleged "software fudging" in the latter (or something.)
We are looking at a JEOL JSM-6490LV and a Hitachi S-3400N specifically, (my manager's choices) and I have seen a CamScan and a few FEI models. My initial impressions are that the CamScan unit is a bit specialized and that the service/support won't be as effective (we are in southern Ontario) and that the FEI machines will be WAY too expensive and more tailored to bio applications / really high-end research. As to the JEOL and Hitachi, I was told the JEOL is slightly better and the company is less likely to disappear (ie, better support and spare parts in the long-term) while the service and support for the Hitachi people is superior, both initially and long-term.
I realize I am asking a great deal, and would appreciate any input into this matter. Has anyone bought or used either of the models listed above? Does anyone have any comments on what we do or do not want? Is Peltier cooling instead of LN2 really a good idea? Any input as to EDS machines and software to buy or to avoid? Thanks for your help.
Gerard Cox
==============================Original Headers============================== 5, 34 -- From gerard.cox-at-goodrich.com Fri Apr 20 12:48:07 2007 5, 34 -- Received: from nhc0sc01.goodrich.com (phx-mxgateway.goodrich.com [63.241.174.69]) 5, 34 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3KHm743005829 5, 34 -- for {Microscopy-at-microscopy.com} ; Fri, 20 Apr 2007 12:48:07 -0500 5, 34 -- Received: from nhc0sc01.goodrich.com (unknown [127.0.0.1]) 5, 34 -- by nhc0sc01.goodrich.com (Symantec Mail Security) with ESMTP id CF2A42842A 5, 34 -- for {Microscopy-at-microscopy.com} ; Fri, 20 Apr 2007 13:48:06 -0400 (EDT) 5, 34 -- X-AuditID: ac150e22-9f039bb00000083a-b8-4628fcd6a15d 5, 34 -- Received: from GR-GWI-WEST-A.goodrich.com (gr-gwi-west-a.goodrich.com [170.126.245.4]) 5, 34 -- by nhc0sc01.goodrich.com (goodrich.com) with ESMTP id AA1D2282AD 5, 34 -- for {Microscopy-at-microscopy.com} ; Fri, 20 Apr 2007 13:48:06 -0400 (EDT) 5, 34 -- Received: from nhc1ex01.goodrich.root.local (localhost [127.0.0.1]) 5, 34 -- by GR-GWI-WEST-A.goodrich.com (8.13.5/8.13.5) with ESMTP id l3KHkf9G024647 5, 34 -- for {Microscopy-at-microscopy.com} ; Fri, 20 Apr 2007 13:48:06 -0400 (EDT) 5, 34 -- Received: from yyz0ex01.goodrich.root.local ([170.126.225.8]) by nhc1ex01.goodrich.root.local with Microsoft SMTPSVC(6.0.3790.1830); 5, 34 -- Fri, 20 Apr 2007 13:47:42 -0400 5, 34 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 5, 34 -- Content-class: urn:content-classes:message 5, 34 -- MIME-Version: 1.0 5, 34 -- Content-Type: text/plain; 5, 34 -- charset="utf-8" 5, 34 -- Subject: New SEM Purchase 5, 34 -- Date: Fri, 20 Apr 2007 13:47:41 -0400 5, 34 -- Message-ID: {FA97C9DCF52E274A94664B4F9FAA9E3DC6546F-at-yyz0ex01.goodrich.root.local} 5, 34 -- X-MS-Has-Attach: 5, 34 -- X-MS-TNEF-Correlator: 5, 34 -- Thread-Topic: New SEM Purchase 5, 34 -- Thread-Index: AceDc/5ZEugnPFylRgaAS8DU5z1Ygg== 5, 34 -- From: "Cox, Gerard" {gerard.cox-at-goodrich.com} 5, 34 -- To: {Microscopy-at-microscopy.com} 5, 34 -- X-OriginalArrivalTime: 20 Apr 2007 17:47:42.0642 (UTC) FILETIME=[FF062920:01C78373] 5, 34 -- X-Brightmail-Tracker: AAAAAA== 5, 34 -- Content-Transfer-Encoding: 8bit 5, 34 -- X-MIME-Autoconverted: from base64 to 8bit by ns.microscopy.com id l3KHm743005829 ==============================End of - Headers==============================
Just a brief question to the list: I've been working on the cooling system to my JSM-840 when an idea struck me. I am having problems because the water pump that I have to circulate the chilled water is not powerful enough to produce the pressure needed to circulate water through the SEMs diffusion pumps. The pump is actually rated for a coolant, though (Antifreeze/DI Water mixture) and I was wondering what you guys think about instead of using water to chill the diffusion pumps and amplifier electronics, using some form of coolant with a lower viscosity than water, which would alleviate my need for such high pressures and let me use the pump that fits the chiller I have.
Can anybody give me an optimal % relative humidity that is recommended for EM rooms? We recently had some failures with formvar grids (we think) due to excessive room humidity when they were being made. I can bring in a de-humidifier, but what would be considered to be optimal?
Teri Johnson, HT(ASCP)QIHC Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, MO 64110
==============================Original Headers============================== 4, 20 -- From tjj-at-stowers-institute.org Fri Apr 20 16:04:09 2007 4, 20 -- Received: from stowers-institute.org (mail.stowers-institute.org [204.56.6.8]) 4, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3KL48qX000489 4, 20 -- for {microscopy-at-microscopy.com} ; Fri, 20 Apr 2007 16:04:08 -0500 4, 20 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 4, 20 -- Content-class: urn:content-classes:message 4, 20 -- MIME-Version: 1.0 4, 20 -- Content-Type: text/plain; 4, 20 -- charset="us-ascii" 4, 20 -- Subject: Recommended humidity level 4, 20 -- Date: Fri, 20 Apr 2007 16:03:58 -0500 4, 20 -- Message-ID: {C28BAF593DC3314E9C0F3A50191C2E7807A3DF2D-at-EXCHKC03.stowers-institute.org} 4, 20 -- X-MS-Has-Attach: 4, 20 -- X-MS-TNEF-Correlator: 4, 20 -- Thread-Topic: Recommended humidity level 4, 20 -- Thread-Index: AceDjyLmsE1bPtgoQnSyl90Uh5X67QAADZUQ 4, 20 -- From: "Johnson, Teri" {TJJ-at-stowers-institute.org} 4, 20 -- To: {microscopy-at-microscopy.com} 4, 20 -- Content-Transfer-Encoding: 8bit 4, 20 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l3KL48qX000489 ==============================End of - Headers==============================
You did not say what brand chiller you have. What flow rate and pressure do you need? For just a diffusion pump (no electronics), I don't think a huge rate is needed. I think you need to set the temperature and have the correct flow rate rather than concentrate on pressure. If you can't get the rate, then that is what you should focus on, IMO.
Haskris and probably others specifically say to NOT use anti-freeze. Some folks survive very well just using distilled water. That has not been my experience. I find that 10% mix of 100% Ethylene Glycol and DI works great. Most chillers use really cheap brass fittings rather than Imperial brass. Consequently, a bad mix of coolant will cause corrosion and also lead to algae growth. The other gotcha are the hoses. Use opaque ones rather than transparent ones. Keeping the light out inhibits algae growth too.
In my Haskris R50 chiller with 5 gallon reservoir, water or the mix results in the same pressure at same flow rate. What will cause pressure to change is a dirty tank filter, or especially the external toilet paper filter (50u fiber mesh).
gary g.
At 12:12 PM 4/20/2007, you wrote:
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If it is for formvar that you wish to keep humidity low, you might consider casting your formvar films within a glove bag flushed with dry nitrogen gas. We use such a system here in our Oregon lab with good success. The bags we use are from I2R and the web address is: http://www.i-2-r.com/glove_bag/l_glove_bags/p_x_r/x_r.htm. I have no financial interest.
Doug
Douglas R. Keene Assistant Investigator Micro-Imaging Center Shriners Hospital for Children 3102 S.W. Sam Jackson Park Road Portland, Oregon 97239 503-221-3434 drk-at-shcc.org
-----Original Message----- X-from: TJJ-at-stowers-institute.org [mailto:TJJ-at-stowers-institute.org] Sent: Friday, April 20, 2007 2:08 PM To: drk-at-SHCC.org
Can anybody give me an optimal % relative humidity that is recommended for EM rooms? We recently had some failures with formvar grids (we think) due to excessive room humidity when they were being made. I can bring in a de-humidifier, but what would be considered to be optimal?
Teri Johnson, HT(ASCP)QIHC Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, MO 64110
==============================Original Headers============================== 4, 20 -- From tjj-at-stowers-institute.org Fri Apr 20 16:04:09 2007 4, 20 -- Received: from stowers-institute.org (mail.stowers-institute.org [204.56.6.8]) 4, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3KL48qX000489 4, 20 -- for {microscopy-at-microscopy.com} ; Fri, 20 Apr 2007 16:04:08 -0500 4, 20 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 4, 20 -- Content-class: urn:content-classes:message 4, 20 -- MIME-Version: 1.0 4, 20 -- Content-Type: text/plain; 4, 20 -- charset="us-ascii" 4, 20 -- Subject: Recommended humidity level 4, 20 -- Date: Fri, 20 Apr 2007 16:03:58 -0500 4, 20 -- Message-ID: {C28BAF593DC3314E9C0F3A50191C2E7807A3DF2D-at-EXCHKC03.stowers-institute.org} 4, 20 -- X-MS-Has-Attach: 4, 20 -- X-MS-TNEF-Correlator: 4, 20 -- Thread-Topic: Recommended humidity level 4, 20 -- Thread-Index: AceDjyLmsE1bPtgoQnSyl90Uh5X67QAADZUQ 4, 20 -- From: "Johnson, Teri" {TJJ-at-stowers-institute.org} 4, 20 -- To: {microscopy-at-microscopy.com} 4, 20 -- Content-Transfer-Encoding: 8bit 4, 20 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l3KL48qX000489 ==============================End of - Headers==============================
==============================Original Headers============================== 14, 22 -- From drk-at-SHCC.org Fri Apr 20 16:36:44 2007 14, 22 -- Received: from mail.SHCC.org (mail.shcc.org [64.213.211.202]) 14, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3KLaiEI013326 14, 22 -- for {Microscopy-at-Microscopy.Com} ; Fri, 20 Apr 2007 16:36:44 -0500 14, 22 -- Received: from DRK2 ([64.213.211.62]) by mail.SHCC.org (PMDF V6.3 #31473) 14, 22 -- with ESMTPA id {0JGT008GWFYUAJ-at-mail.SHCC.org} for Microscopy-at-Microscopy.Com; 14, 22 -- Fri, 20 Apr 2007 14:35:18 -0700 (PDT) 14, 22 -- Date: Fri, 20 Apr 2007 14:37:27 -0700 14, 22 -- From: Doug Keene {drk-at-SHCC.org} 14, 22 -- Subject: RE: [Microscopy] Recommended humidity level 14, 22 -- In-reply-to: {200704202107.l3KL7XLD007804-at-ns.microscopy.com} 14, 22 -- Sender: drk-at-SHCC.org 14, 22 -- To: Microscopy-at-Microscopy.Com 14, 22 -- Reply-to: drk-at-SHCC.org 14, 22 -- Message-id: {0JGT008GXFYUAJ-at-mail.SHCC.org} 14, 22 -- Organization: Shriners Hospitals for Children 14, 22 -- MIME-version: 1.0 14, 22 -- X-MIMEOLE: Produced By Microsoft MimeOLE V6.00.2900.3028 14, 22 -- X-Mailer: Microsoft Office Outlook, Build 11.0.5510 14, 22 -- Content-type: text/plain; charset=us-ascii 14, 22 -- Content-transfer-encoding: 7BIT 14, 22 -- thread-index: AceDj8Hmj6FwMw8kREWYusgUcEBeAAAA23cg ==============================End of - Headers==============================
Hi Justin, Does JEOL list a spec, for the flow rate? Too much may give you excess vibration.
Could the lines/pump have some blockage?
Restricting the outlet, a little, will increase the backpressure, it might be enough to trip the interlock, if that's shutting things off.
I've added up to 5% ETHYLENE GLYCOL to recirculating cooling systems (EBEAM and SEM and TEM) with no long-term deleterious effects. Not sure how much that will change the viscosity.
I've always been advised against running DI water alone as it is ion hungry and may leach metal pipes. I've always used grocery store distilled water.
Best of Luck.
Ed Basgall, PhD Irvine, CA 92612
email: ejb1176-at-gmail.com
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On Apr 20, 2007, at 2:04 PM, TJJ-at-stowers-institute.org wrote:
} Can anybody give me an optimal % relative humidity that is recommended } for EM rooms? We recently had some failures with formvar grids (we } think) due to excessive room humidity when they were being made. I can } bring in a de-humidifier, but what would be considered to be optimal? } Dear Teri, You need not only to be concerned about the formvar, but you also do not want condensation on the column or electronics. Too low a humidity will also allow the generation of static electricity. About 40% RH is a very good value, and the maximum permissible depends on the difference in temperature between the room and the scope and is usually about 60%. 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, 24 -- From tivol-at-caltech.edu Fri Apr 20 17:19:29 2007 4, 24 -- Received: from outgoing-mail.its.caltech.edu (outgoing-mail.its.caltech.edu [131.215.239.19]) 4, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3KMJTYb015114 4, 24 -- for {microscopy-at-msa.microscopy.com} ; Fri, 20 Apr 2007 17:19:29 -0500 4, 24 -- Received: from water-dog.its.caltech.edu (water-dog [192.168.1.26]) 4, 24 -- by wood-ox-postvirus (Postfix) with ESMTP id 4E1AF2F120 4, 24 -- for {microscopy-at-msa.microscopy.com} ; Fri, 20 Apr 2007 15:19:18 -0700 (PDT) 4, 24 -- Received: from [192.168.159.233] (jpix-01.caltech.edu [131.215.2.133]) 4, 24 -- (using TLSv1 with cipher RC4-SHA (128/128 bits)) 4, 24 -- (No client certificate requested) 4, 24 -- by water-ox.its.caltech.edu (Postfix) with ESMTP id 5D6002EF43 4, 24 -- for {microscopy-at-msa.microscopy.com} ; Fri, 20 Apr 2007 15:19:16 -0700 (PDT) 4, 24 -- Mime-Version: 1.0 (Apple Message framework v624) 4, 24 -- In-Reply-To: {200704202104.l3KL4HOb000636-at-ns.microscopy.com} 4, 24 -- References: {200704202104.l3KL4HOb000636-at-ns.microscopy.com} 4, 24 -- Content-Type: text/plain; charset=US-ASCII; format=flowed 4, 24 -- Message-Id: {b41796fa0e13270b6f2fd4c8b3342e1d-at-caltech.edu} 4, 24 -- Content-Transfer-Encoding: 7bit 4, 24 -- From: Bill Tivol {tivol-at-caltech.edu} 4, 24 -- Subject: Re: [Microscopy] Recommended humidity level 4, 24 -- Date: Fri, 20 Apr 2007 15:30:10 -0700 4, 24 -- To: microscopy-at-msa.microscopy.com 4, 24 -- X-Mailer: Apple Mail (2.624) 4, 24 -- X-Spam-Scanned: at Caltech-ITS on water-dog by amavisd-2.3.3 ==============================End of - Headers==============================
Sorry, I wasn't very clear about the chiller before. It's not a name brand chiller- it's actually a part from a larger system, but I don't know what that larger system is- I bought it locally as industrial surplus. Here's what I know about the recirculator system:
Pump: Iwaki Magnetic drive pump. 12 L/min. Max. head 2.1m.
There is a heat exchanger unit and all of the condenser piping necessary to chill the water, and it can divide the output a maximum of three ways, so I can have the electronics set on a different water circuit than the diffusion pumps. When I say that the pressure is not enough to push the water through, I mean that the pump pumps, but very little water is pushed through, and it is shutting down because of thermal overload. I know that the JEOL manual specifies 5 L/min. at 12-36 PSI, but I'm not sure how that translates from "Max head" to PSI. The pump's flow rate is more than enough to handle it, but it isn't making it through. That's why I was thinking about lower viscosity fluids, which should flush through at a higher flow rate and a lower PSI.
--Justin.
On 4/20/07, 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 } ---------------------------------------------------------------------------- } } Just a brief question to the list: I've been working on the cooling } system to my JSM-840 when an idea struck me. I am having problems } because the water pump that I have to circulate the chilled water is } not powerful enough to produce the pressure needed to circulate water } through the SEMs diffusion pumps. The pump is actually rated for a } coolant, though (Antifreeze/DI Water mixture) and I was wondering } what you guys think about instead of using water to chill the } diffusion pumps and amplifier electronics, using some form of coolant } with a lower viscosity than water, which would alleviate my need for } such high pressures and let me use the pump that fits the chiller I } have. } } --Justin A. Kraft } } ==============================Original Headers============================== } 2, 26 -- From kraftpiano-at-gmail.com Fri Apr 20 15:10:08 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 l3KKA6rl020523 } 2, 26 -- for {microscopy-at-microscopy.com} ; Fri, 20 Apr 2007 15:10:07 -0500 } 2, 26 -- Received: by wr-out-0506.google.com with SMTP id i31so1048787wra } 2, 26 -- for {microscopy-at-microscopy.com} ; Fri, 20 Apr 2007 13:10:06 -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=V6b5XTIKIpdd6yYdQ6xnjSvMcCaYrgSlyHfpsumqQ6ZTo4rgHH4TLCz9a7EPstJ0IBPup7o+/fc/quOtZWUR+gHHAn/qtAC3EShd3fp2pEfrggcJMZcTuTgwLcRyyH/ok8PIJjj954430XfVnymNnYrnD3TIaOFlz1OJy9v8j/U= } 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=JYwtFEuBzBh8ycTaM6mQ+VYn9+ea0ojpzxZdvjQDsAhXcyralyl62mWg2ZwSPoQeJC9JsObe0K6/DIijuQsW0ek4s5lkoZGp48IwujXLCPnYfJcx2VJJLgBnSzdg7zImpfBwndCEogJKWtsHDhnzfv7Zgvz6mM2m78jw+cph2hQ= } 2, 26 -- Received: by 10.114.57.1 with SMTP id f1mr1412835waa.1177099805596; } 2, 26 -- Fri, 20 Apr 2007 13:10:05 -0700 (PDT) } 2, 26 -- Received: by 10.114.79.12 with HTTP; Fri, 20 Apr 2007 13:10:05 -0700 (PDT) } 2, 26 -- Message-ID: {25e2b0d20704201310u2cfc1b75q670356e82dea57f9-at-mail.gmail.com} } 2, 26 -- Date: Fri, 20 Apr 2007 16:10:05 -0400 } 2, 26 -- From: "Justin Kraft" {kraftpiano-at-gmail.com} } 2, 26 -- To: microscopy-at-microscopy.com } 2, 26 -- Subject: SEM: Replacing diffusion pump water cycle with coolant. } 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============================== }
ditto that- 30 psi is enough, unless cooling system is clogged. If it is clogged, then concentrate your efforts on unclogging the cooling system, rather than increasing pressure. The highest water pressure that I can imagine being required for any SEM/TEM is in mid-40 psi. I typically set much lesser pressure, between 25 and 35 psi. Compensating poor flow in a clogged system with higher pressure is a bad habit. Plus, high pressure erodes rubber and plastic components of the cooling system, and that alone accelerates formation of clogs. Especially with old instrument.
I suggest to have at least 1 flowmeter with flow control (it is cheap). And pressure reducer with bypass line for the water pump - every decent chiller has that anyway. Pump itself shall be capable of at least 100 psi. Not for everyday operation, but for troubleshooting, especially if you have old instrument. You can easily detect presence of a clog (if not location...), by adjusting and monitoring pressure and flow rate. More than one flowmeter is good to have if cooling system has parallel branches.
Likely places for clogs in JEOL-840 are: quick connections, lower (hottest) water line coils on both ODPs, and of course filters if any are present. Less likely places are electronics and obj. lens. But this is never certain.
To remove clog, reverse the direction of water flow, or use small amount water with compressed air in reversed direction. But first disconnect various parts of cooling circuit, and determine where the clog is located. If you reverse the whole thing at once, clog might get worse and harder to remove. You will need a variety of hoses and fittings to do this work efficiently.
Flow rates at room T~20C and water T~17C: 1gpm is great, 0.5gpm is OK, 0.2 gpm is critically low, below that expect thermal protection to trip at any moment.
Vitaly Feingold SIA 2773 Heath Lane Duluth GA 30096 Ph. 770-232-7785 Fax 770-232-1791 www.sia-cam.com ----- Original Message ----- X-from: {gary-at-gaugler.com} To: {vitalylazar-at-att.net} Sent: Friday, April 20, 2007 5:37 PM
==============================Original Headers============================== 9, 24 -- From vitalylazar-at-att.net Fri Apr 20 18:05:04 2007 9, 24 -- Received: from mtiwmhc12.worldnet.att.net (mtiwmhc12.worldnet.att.net [204.127.131.116]) 9, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3KN54LK006441 9, 24 -- for {Microscopy-at-microscopy.com} ; Fri, 20 Apr 2007 18:05:04 -0500 9, 24 -- Received: from siap6segs20pa8 (h-68-165-14-60.atlngahp.dynamic.covad.net[68.165.14.60]) 9, 24 -- by worldnet.att.net (mtiwmhc12) with SMTP 9, 24 -- id {20070420230503112004gh36e} ; Fri, 20 Apr 2007 23:05:03 +0000 9, 24 -- Message-ID: {004a01c783a0$54872ee0$6601a8c0-at-siap6segs20pa8} 9, 24 -- From: "Vitaly Feingold" {vitalylazar-at-att.net} 9, 24 -- To: "Microscopy Listserver" {Microscopy-at-microscopy.com} , 9, 24 -- "Gary Gaugler" {gary-at-gaugler.com} 9, 24 -- References: {200704202137.l3KLbj9U017071-at-ns.microscopy.com} 9, 24 -- Subject: Re: [Microscopy] Re: SEM: Replacing diffusion pump water cycle 9, 24 -- Date: Fri, 20 Apr 2007 19:04:09 -0400 9, 24 -- MIME-Version: 1.0 9, 24 -- Content-Type: text/plain; 9, 24 -- format=flowed; 9, 24 -- charset="iso-8859-1"; 9, 24 -- reply-type=original 9, 24 -- Content-Transfer-Encoding: 7bit 9, 24 -- X-Priority: 3 9, 24 -- X-MSMail-Priority: Normal 9, 24 -- X-Mailer: Microsoft Outlook Express 6.00.2900.3028 9, 24 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.3028 ==============================End of - Headers==============================
Fracture method will not work on this sample. Stainless steel and copper are very ductile and will bend and neck down before breaking. These metals are not made brittle by cold temperatures.
Sample preparation, i.e. good polishing methods is the key to this evaluation. This combination can be polished mechanically, but a good procedure and technique will be required.
-- Larry D. Hanke, P.E. Materials Evaluation and Engineering, Inc. Practical Solutions Through Technology and Innovation http://www.mee-inc.com (763) 449-8870
==============================Original Headers============================== 4, 25 -- From hanke-at-mee-inc.com Fri Apr 20 18:42:58 2007 4, 25 -- Received: from mail.namisolutions.com (mail.namisolutions.com [12.40.181.38]) 4, 25 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3KNgwSX018376 4, 25 -- for {microscopy-at-microscopy.com} ; Fri, 20 Apr 2007 18:42:58 -0500 4, 25 -- Received: (qmail 29139 invoked by uid 508); 20 Apr 2007 18:42:58 -0500 4, 25 -- Received: from 216.14.180.82 by mail.namisolutions.com (envelope-from {hanke-at-mee-inc.com} , uid 507) with qmail-scanner-1.24-st-qms 4, 25 -- (clamdscan: 0.80/650. spamassassin: 3.0.2. perlscan: 1.24-st-qms. 4, 25 -- Clear:RC:1(216.14.180.82):. 4, 25 -- Processed in 0.105867 secs); 20 Apr 2007 23:42:58 -0000 4, 25 -- X-Antivirus-NAMISOLUTIONS-Mail-From: hanke-at-mee-inc.com via mail.namisolutions.com 4, 25 -- X-Antivirus-NAMISOLUTIONS: 1.24-st-qms (Clear:RC:1(216.14.180.82):. Processed in 0.105867 secs Process 29132) 4, 25 -- Received: from unknown (HELO ?192.168.1.4?) (216.14.180.82) 4, 25 -- by mail.namisolutions.com with SMTP; 20 Apr 2007 18:42:57 -0500 4, 25 -- Message-ID: {46294FE9.1010103-at-mee-inc.com} 4, 25 -- Date: Fri, 20 Apr 2007 18:42:33 -0500 4, 25 -- From: Larry Hanke {hanke-at-mee-inc.com} 4, 25 -- User-Agent: Mozilla Thunderbird 1.0.6 (Windows/20050716) 4, 25 -- X-Accept-Language: en-us, en 4, 25 -- MIME-Version: 1.0 4, 25 -- To: microscopy-at-microscopy.com 4, 25 -- Subject: Re: cross section sample preparation 4, 25 -- References: {200704201123.l3KBNUkd021202-at-ns.microscopy.com} 4, 25 -- In-Reply-To: {200704201123.l3KBNUkd021202-at-ns.microscopy.com} 4, 25 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 4, 25 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
The reason NOT to post to the list is that you get volumes of "Out Of Office" messages. This is the same as when you make a new posting. Most of the defunct messages are trapped by Eudora but quite a few still get though....sigh.
Some do not have this subject but say that they are gone from xx to yy.
gary g.
At 03:50 PM 4/19/2007, you wrote: } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
==============================Original Headers============================== 7, 20 -- From gary-at-gaugler.com Fri Apr 20 20:49:45 2007 7, 20 -- Received: from qsmtp4.mc.surewest.net (qsmtp.mc.surewest.net [66.60.130.145]) 7, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id l3L1niqX031833 7, 20 -- for {microscopy-at-microscopy.com} ; Fri, 20 Apr 2007 20:49:45 -0500 7, 20 -- Message-Id: {200704210149.l3L1niqX031833-at-ns.microscopy.com} 7, 20 -- Received: (qmail 25911 invoked from network); 20 Apr 2007 18:49:44 -0700 7, 20 -- Received: by simscan 1.1.0 ppid: 25908, pid: 25909, t: 0.1368s 7, 20 -- scanners: regex: 1.1.0 attach: 1.1.0 7, 20 -- Received: from unknown (HELO ivan.gaugler.com) (66.60.171.211) 7, 20 -- by qsmtp4 with SMTP; 20 Apr 2007 18:49:44 -0700 7, 20 -- X-Mailer: QUALCOMM Windows Eudora Version 7.1.0.9 7, 20 -- Date: Fri, 20 Apr 2007 18:49:48 -0800 7, 20 -- To: tom-at-tomkaye.com 7, 20 -- From: Gary Gaugler {gary-at-gaugler.com} 7, 20 -- Subject: Re: [Microscopy] 2nd for posting to the list 7, 20 -- Cc: MSA listserver {microscopy-at-microscopy.com} 7, 20 -- In-Reply-To: {200704192350.l3JNo7lp017141-at-ns.microscopy.com} 7, 20 -- References: {200704192350.l3JNo7lp017141-at-ns.microscopy.com} 7, 20 -- Mime-Version: 1.0 7, 20 -- Content-Type: text/plain; charset=us-ascii; format=flowed; x-avg-checked=avg-ok-2818210D ==============================End of - Headers==============================
Dear Alvaro, interesting question.... Normally (e.g. as I use it sometimes for negative staining of virus particles) you will have at least two negative staining solutions: i) hydrous PTA (1, 2 or other percentage) ii) hydrous uranylacetate 1-4%....... for (small) lipoproteins (esp. for HDL-like) I am not quite sure wether simple saying so or giving advice in that way is helpful.
As I understand negative staing, also (a) positive result(s)will depend on pH (and therefore [with what ingredients] buffering the solution eventually to which "special" pH), and (absorption/adhesion) technique of specimen preparation. I don't think your carbon-coated formvar-filmed copper grids are a problem....but,. if I remember correctly, there are several (also older) papers/articles on negative staining techniques for e.g. liposomes (for an imagination on such try search phrase } } "negative staining","liporotein" { {) reporting some very specific caudeles for adhesion of lipoprotein material....(for instance: see*) below) Some prefer also prefixation of mixed particles with hydrous Osmium tetroxide before negative staining with the heavy metal solution, sometimes only hydrous, somtetimes with an additive (like sucrose)..... so it may depend also on the type of material you'd like to have demonstrated and wether you are able to get the (macro-)molecule adhering in a stabilized form to the formvar-carbon-surface of the grid or not (cf. *)
Once determined [by trial&error (perhaps, unluckily)] how to proceed with the grids' properties = hydrophilization and immediate use or storage for months...see *), adhesion time/technique, eventually prestabilization of the specimen by additional fixation, the right negative staining solution (also in terms of i) element [PTA, UO2, AmmMolybdat,Uranylformate, etc.], ii)concentration, iii) pH of the solution, iv) specific additives necessary?) the technique itself yields reliable and rapid rsults, even within 5-10 minutes (as I have read in some papers) ==} that's what I would like to wish you in overcoming your problem.
Best regards Wolfgang Muss, Salzburg, AUSTRIA
cf: *) Original paper in: J Lipid Res. 1980 Nov;21(8):981-92. Unilamellar liposomes made with the French pressure cell: a simple preparative and semiquantitative technique. Hamilton RL Jr, Goerke J, Guo LS, Williams MC,Havel RJ. Ex Material & Methods-section: Negative staining of liposomes often causes artifactual images resulting from disruption of small liposomes that subsequently form larger multilamellar structures. Although the mechanism of this process is not understood, it may be caused in part by the intense hydrophobicity of freshly evaporated carbon on grid surfaces. Our attempts to modify the surface film of freshly prepared carbon-coated grids (200-300 mesh copper grids, Fullum, Schenectady,NY) by glow discharge, polylysine, or addition of albumin to sample or grid did not prove satisfactory. However, we have learned empirically that parlodioncovered grids with carbon films made with a Siemens evaporator (VI3 G 500) permit even spreading of liposomes without apparent structural changes, provided that the grids have been aged 6- 12 months on the shelf. Carbon rods for filming were obtained from Ringsdorff-Herke (Spektral Kohlen, Bonn-Bad Godesberg, West Germany). With such “matured” carbon-coated grids, liposomes were prepared for electron microscopy with 2% potassium phosphotungstate at pH 6.45-6.5 by the drop procedure described previously (1 1). We found that the lipid concentration in the sample was also important for preparing uniform spreads. Optimal results were more consistently obtained with samples containing 1- 10 mg/ml PC. Improved results can be obtained for more dilute samples by increasing contact time of the sample on the grid to 2-3 min, and by using a tighter 400 mesh grid.......
Zitat von alvarobq-at-fcien.edu.uy:
} } } } ---------------------------------------------------------------------------- } 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 (alvarobq-at-fcien.edu.uy) from } http://www.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on } Friday, September 15, 2006 at 08:40:57 } --------------------------------------------------------------------------- } } Email: alvarobq-at-fcien.edu.uy } Name: Alvaro D. Olivera } } Organization: Science University } } Education: Undergraduate College } } Location: Montevideo - Uruguay } } Question: } } I'm TEM technician and advanced undergraduate in Biochemistry. } I'm trying negative staining whith a small lipo protein HDL like. I'm } using CARBON-FORMVAR COATED Cu GRIDS and PTA 2% and 3%. I can't see it. } What do you suggest? } } Many thanks, Alvaro. } } --------------------------------------------------------------------------- } } ==============================Original Headers============================== } 9, 12 -- From zaluzec-at-ultra5.microscopy.com Fri Sep 15 08:47:52 2006 } 9, 12 -- Received: from [206.69.208.22] (mac22.zaluzec.com [206.69.208.22]) } 9, 12 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id } k8FDlpKs009958 } 9, 12 -- for {microscopy-at-microscopy.com} ; Fri, 15 Sep 2006 08:47:51 -0500 } 9, 12 -- Mime-Version: 1.0 } 9, 12 -- X-Sender: zaluzec-at-ultra5.microscopy.com (Unverified) } 9, 12 -- Message-Id: {p06110406c1305f547915-at-[206.69.208.22]} } 9, 12 -- Date: Fri, 15 Sep 2006 08:47:50 -0500 } 9, 12 -- To: microscopy-at-microscopy.com } 9, 12 -- From: alvarobq-at-fcien.edu.uy (by way of Ask-A-Microscopist) } 9, 12 -- Subject: AskAMicroscopist: negative staining whith a small lipo } protein } 9, 12 -- Content-Type: text/plain; charset="us-ascii" } ==============================End of - Headers============================== }
==============================Original Headers============================== 13, 37 -- From W.Muss-at-salk.at Sat Apr 21 06:14:20 2007 13, 37 -- Received: from hermes.salk.at (hermes.salk.at [193.170.167.9]) 13, 37 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3LBEJVe019352 13, 37 -- for {microscopy-at-microscopy.com} ; Sat, 21 Apr 2007 06:14:19 -0500 13, 37 -- Received: from localhost (localhost [127.0.0.1]) 13, 37 -- by hermes.salk.at (Postfix) with ESMTP id B683EC386F; 13, 37 -- Sat, 21 Apr 2007 13:14:17 +0200 (CEST) 13, 37 -- X-Virii-Scanned: Kaspersky Antivirus at salk.at 13, 37 -- Received: from hermes.salk.at ([127.0.0.1]) 13, 37 -- by localhost (n1ex218.lks.local [127.0.0.1]) (amavisd-new, port 10024) 13, 37 -- with ESMTP id zNBlE5G61yau; Sat, 21 Apr 2007 13:14:17 +0200 (CEST) 13, 37 -- Received: from hermes.lks.at (hermes.salk.at [192.168.13.210]) 13, 37 -- by hermes.salk.at (Postfix) with ESMTP id 4FAEDC3844; 13, 37 -- Sat, 21 Apr 2007 13:14:17 +0200 (CEST) 13, 37 -- Received: from localhost (localhost [127.0.0.1]) 13, 37 -- by hermes.lks.at (Postfix) with ESMTP id 262F35A9044; 13, 37 -- Sat, 21 Apr 2007 13:14:17 +0200 (CEST) 13, 37 -- Received: from hermes.lks.at ([127.0.0.1]) 13, 37 -- by localhost (hermes.lks.local [127.0.0.1]) (amavisd-new, port 10024) 13, 37 -- with ESMTP id 62675-03; Sat, 21 Apr 2007 13:14:16 +0200 (CEST) 13, 37 -- Received: from salk.at (webmail.salk.at [192.168.13.209]) 13, 37 -- by hermes.lks.at (Postfix) with ESMTP id A02E55A9041; 13, 37 -- Sat, 21 Apr 2007 13:14:16 +0200 (CEST) 13, 37 -- Received: from m069p011.adsl.highway.telekom.at (m069p011.adsl.highway.telekom.at [62.47.176.139]) 13, 37 -- by webmail.salk.at (IMP) with HTTP 13, 37 -- for {pawma-at-192.168.13.210} ; Sat, 21 Apr 2007 13:14:16 +0200 13, 37 -- Message-ID: {1177154056.4629f20873ac5-at-webmail.salk.at} 13, 37 -- Date: Sat, 21 Apr 2007 13:14:16 +0200 13, 37 -- From: Wolfgang Muss {W.Muss-at-salk.at} 13, 37 -- To: alvarobq-at-fcien.edu.uy, microscopy-at-microscopy.com 13, 37 -- Subject: [Microscopy]Re:AskAMicroscopist: negative staining whith a small lipo protein 13, 37 -- MIME-Version: 1.0 13, 37 -- Content-Type: text/plain; charset=ISO-8859-1 13, 37 -- Content-Transfer-Encoding: 8bit 13, 37 -- User-Agent: Internet Messaging Program (IMP) 3.2.8 13, 37 -- X-Virii-Scanned: by Sophos/ClamAV at salk.at 13, 37 -- X-Scanned-By: SALK-Content-Filter ==============================End of - Headers==============================
gary-at-gaugler.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 } ---------------------------------------------------------------------------- } } The reason NOT to post to the list is that you get } volumes of "Out Of Office" messages. This is the } same as when you make a new posting. Most of the } defunct messages are trapped by Eudora but quite } a few still get though....sigh. } } Some do not have this subject but say that they } are gone from xx to yy. } } gary g. } } } } At 03:50 PM 4/19/2007, 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 } } ---------------------------------------------------------------------------- } } } } } } I would like to 2nd the idea that replying to the list is a good thing. This } } is my only resource for such information so I would rather delete than miss } } an opportunity to learn something. } } } } Thanks! } } } } Tom Kaye } } } } Not affiliated with anything. } } } } -----Original Message----- } } X-from: cervantes-at-bendres.com [mailto:cervantes-at-bendres.com] } } Sent: Thursday, April 19, 2007 6:02 PM } } To: tom-at-tomkaye.com } } Subject: [Microscopy] RE: stabilization of membranes } } } } } } } } ---------------------------------------------------------------------------- } } 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 } } ---------------------------------------------------------------------------- } } } } Did I miss the replies to this query somehow? Is there a technical } } problem, or were the replies private? Would it be possible to post } } them? } } } } Let's not be shy (if that's it) about posting to the whole list. I've } } gotten so much out of this resource (especially lately). I've had to } } re-post replies to my query for others who have had the same question } } and hadn't gotten the messages. } } } } Thanks for keeping the information flowing; it's invaluable. } } Jessica Cervantes } } Bend Research Inc } } Bend, OR } } } } -----Original Message----- } } X-from: drk-at-SHCC.org [mailto:drk-at-SHCC.org] } } Sent: Wednesday, April 18, 2007 10:49 AM } } To: Cervantes, Jessica } } Subject: [Microscopy] stabilization ofmembranes } } } } ------------------------------------------------------------------------ } } ---- } } 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 } } ------------------------------------------------------------------------ } } ---- } } } } Fellow Microscopists, } } } } } } A few days ago I posted a query for suggestions on how to stabilize } } lysosomal membranes in cultured cells prior to immunocytochemistry. We } } have } } tried several of the tips but without success. Thanks to those who } } replied! } } We have been following the condition of the lysosomes using a GFP tag } } that } } we know to be specifically compartmentalized. We note that following } } fixation and a rinse in TRIS-Glycine the GFP is still compartmentalized. } } However, perhaps in 90% EtOH and definitely in 1:1 LR White:90% EtOH we } } see } } the GFP moves from the lysosomes to the cytoplasm and nucleus. It seems } } that } } ethanol and certainly LR White contribute to the deterioration of } } membranes. } } Can anyone suggest an embedding media that might be a little gentler on } } membranes and still possibly work for surface label immunocytochemistry? } } We } } are thinking of trying Nanoplast, a water soluble media but any } } suggestions } } are welcome. } } } } } } Thanks! Doug } } } } Douglas R. Keene } } Assistant Investigator } } Micro-Imaging Center } } Shriners Hospital for Children } } 3102 S.W. Sam Jackson Park Road } } Portland, Oregon 97239 } } 503-221-3434 } } drk-at-shcc.org } } } } } } ==============================Original } } Headers============================== } } 7, 22 -- From drk-at-SHCC.org Wed Apr 18 12:43:18 2007 } } 7, 22 -- Received: from mail.SHCC.org (mail.shcc.org [64.213.211.202]) } } 7, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with } } ESMTP id l3IHhITP002098 } } 7, 22 -- for {Microscopy-at-Microscopy.Com} ; Wed, 18 Apr 2007 } } 12:43:18 -0500 } } 7, 22 -- Received: from DRK2 ([64.213.211.62]) by mail.SHCC.org (PMDF } } V6.3 #31473) } } 7, 22 -- with ESMTPA id {0JGP0029FFTXS8-at-mail.SHCC.org} for } } Microscopy-at-Microscopy.Com; } } 7, 22 -- Wed, 18 Apr 2007 10:41:57 -0700 (PDT) } } 7, 22 -- Date: Wed, 18 Apr 2007 10:43:58 -0700 } } 7, 22 -- From: Doug Keene {drk-at-SHCC.org} } } 7, 22 -- Subject: stabilization ofmembranes } } 7, 22 -- Sender: drk-at-SHCC.org } } 7, 22 -- To: Microscopy-at-Microscopy.Com } } 7, 22 -- Cc: drk-at-SHCC.org } } 7, 22 -- Reply-to: drk-at-SHCC.org } } 7, 22 -- Message-id: {0JGP0029GFTXS8-at-mail.SHCC.org} } } 7, 22 -- Organization: Shriners Hospitals for Children } } 7, 22 -- MIME-version: 1.0 } } 7, 22 -- X-MIMEOLE: Produced By Microsoft MimeOLE V6.00.2900.3028 } } 7, 22 -- X-Mailer: Microsoft Office Outlook, Build 11.0.5510 } } 7, 22 -- Content-type: text/plain; charset=us-ascii } } 7, 22 -- Content-transfer-encoding: 7BIT } } 7, 22 -- Thread-Index: AceB4SRWHFc2k9QuQCCG4KbQ3H8v4A== } } ==============================End of - } } Headers============================== } } } } } } ==============================Original Headers============================== } } 13, 16 -- From cervantes-at-bendres.com Thu Apr 19 17:57:57 2007 } } 13, 16 -- Received: from BRIEX04A.bri.local (bc161112.bendcable.com } } [216.228.161.112]) } } 13, 16 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id } } l3JMvuoK002012 } } 13, 16 -- for {Microscopy-at-microscopy.com} ; Thu, 19 Apr 2007 } } 17:57:56 -0500 } } 13, 16 -- MIME-Version: 1.0 } } 13, 16 -- Content-Type: text/plain; } } 13, 16 -- charset="us-ascii" } } 13, 16 -- Subject: RE: [Microscopy] stabilization of membranes } } 13, 16 -- Date: Thu, 19 Apr 2007 15:57:55 -0700 } } 13, 16 -- Message-ID: {8943D65F9AD70E4488AD6DE09F15088768C47F-at-BRIEX04A} } } 13, 16 -- In-Reply-To: {200704181748.l3IHmsrE012175-at-ns.microscopy.com} } } 13, 16 -- References: {200704181748.l3IHmsrE012175-at-ns.microscopy.com} } } 13, 16 -- From: "Cervantes, Jessica" {cervantes-at-bendres.com} } } 13, 16 -- To: {Microscopy-at-microscopy.com} } } 13, 16 -- Content-Transfer-Encoding: 8bit } } 13, 16 -- X-MIME-Autoconverted: from quoted-printable to 8bit by } } ns.microscopy.com id l3JMvuoK002012 } } ==============================End of - Headers============================== } } } } } } ==============================Original Headers============================== } } 25, 20 -- From tom-at-tomkaye.com Thu Apr 19 18:47:51 2007 } } 25, 20 -- Received: from tomkaye.com (mxgateway.techpro.com [64.4.207.8]) } } 25, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with } } ESMTP id l3JNlodW014261 } } 25, 20 -- for {microscopy-at-microscopy.com} ; Thu, 19 Apr 2007 } } 18:47:51 -0500 } } 25, 20 -- Received: from tk7c797a275194 [24.15.58.103] by } } tomkaye.com with ESMTP } } 25, 20 -- (SMTPD32-8.14) id AFAD3312012A; Thu, 19 Apr 2007 18:47:57 -0500 } } 25, 20 -- From: "Tom Kaye" {tom-at-tomkaye.com} } } 25, 20 -- To: {microscopy-at-microscopy.com} } } 25, 20 -- Subject: 2nd for posting to the list } } 25, 20 -- Date: Thu, 19 Apr 2007 18:48:03 -0500 } } 25, 20 -- Message-ID: {GOEAIJOJDGCJBPIFHOBGKEPBOOAA.tom-at-tomkaye.com} } } 25, 20 -- MIME-Version: 1.0 } } 25, 20 -- Content-Type: text/plain; } } 25, 20 -- charset="iso-8859-1" } } 25, 20 -- Content-Transfer-Encoding: 7bit } } 25, 20 -- X-Priority: 3 (Normal) } } 25, 20 -- X-MSMail-Priority: Normal } } 25, 20 -- X-Mailer: Microsoft Outlook IMO, Build 9.0.2416 (9.0.2911.0) } } 25, 20 -- x-mimeole: Produced By Microsoft MimeOLE V6.00.2900.3028 } } 25, 20 -- Importance: Normal } } ==============================End of - Headers============================== } } } } } ==============================Original Headers============================== } 7, 20 -- From gary-at-gaugler.com Fri Apr 20 20:49:45 2007 } 7, 20 -- Received: from qsmtp4.mc.surewest.net (qsmtp.mc.surewest.net [66.60.130.145]) } 7, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id l3L1niqX031833 } 7, 20 -- for {microscopy-at-microscopy.com} ; 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==============================Original Headers============================== 4, 19 -- From randerson20-at-tampabay.rr.com Sat Apr 21 06:36:32 2007 4, 19 -- Received: from ms-smtp-05.tampabay.rr.com (ms-smtp-05.tampabay.rr.com [65.32.5.135]) 4, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3LBaWBQ031114 4, 19 -- for {Microscopy-at-Microscopy.Com} ; Sat, 21 Apr 2007 06:36:32 -0500 4, 19 -- Received: from [127.0.0.1] (rrcs-24-73-73-214.se.biz.rr.com [24.73.73.214]) 4, 19 -- by ms-smtp-05.tampabay.rr.com (8.13.6/8.13.6) with ESMTP id l3LBaT31019574; 4, 19 -- Sat, 21 Apr 2007 07:36:30 -0400 (EDT) 4, 19 -- Message-ID: {4629F73C.5030809-at-tampabay.rr.com} 4, 19 -- Date: Sat, 21 Apr 2007 07:36:28 -0400 4, 19 -- From: Ronald Anderson {randerson20-at-tampabay.rr.com} 4, 19 -- User-Agent: Thunderbird 1.5.0.10 (Windows/20070221) 4, 19 -- MIME-Version: 1.0 4, 19 -- To: gary-at-gaugler.com, Listserver {Microscopy-at-Microscopy.Com} 4, 19 -- Subject: Re: [Microscopy] Re: 2nd for posting to the list 4, 19 -- References: {200704210149.l3L1ntal032017-at-ns.microscopy.com} 4, 19 -- In-Reply-To: {200704210149.l3L1ntal032017-at-ns.microscopy.com} 4, 19 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 4, 19 -- Content-Transfer-Encoding: 7bit 4, 19 -- X-Virus-Scanned: Symantec AntiVirus Scan Engine ==============================End of - Headers==============================
Replying to the list is not only appropriate, but should be done in almost all cases. The obvious exception I could see is when a private reply is appropriate, or a vendor is discussing a reply to a query with detailed product information about a commerical item to an individuals request.
Remember the primary purpose of this list is to share our communal expertise, if you post messages privately then this knowledge distribution is curtailed. Of course, some people post summaries of replies to their questions. This is a good procedure, and is also suggested in the FAQ, and I would encourage people who receive alot of private replies to do this as a courtesy to the community. Consider it part of the way you pay the astronomical annual fees charged to each of you for being a member of this forum.
FYI, there is nothing I can do to stop "out of the office" type replies, since those come from the individuals and never pass through the listserver filters. If you read the Listserver FAQ, I request that if you are unavailable then you should unsubscribe from the list and then resubscribe when you return. There are alot of people that do this (THANK YOU), but obviously some do not.
In principle you won't miss anything, if you unsubscribe for a short while, as all the messages are archived, however, there are clearly people who choose not to follow this request for various reasons.
An alternative should you be wary of missing something, or if your company policy requires you to use Out of the Office messages, would be to have 2 addresses. One for your official mail (to which you could add the Out of the Office reply), while a second which is only used for Listserver traffic. This would be the logical approach for those subscribers who use the Out of the Office message frequently.
As Gary G. mentioned most Email programs today allow you to filter mail by topic, and I certainly, also use this mechanism to refine the distribution of all Emails I personally get into appropriate folders. If you do this then the issue becomes relatively minor, considering the proponderance of SPAM/JUNK mail.
If it is not obvious to you by now all Listserver Email has the following text in the subject line [Microscopy] that was done in order to allow you to create a filter to put all listserver Email into a folder.
Fortunately for all of you the primary filters on the subscription lists now catch nearly all of the junk/spam mail which is sent to "microscopy-at-". For the record, long ago spam exceeded 500 messages/day, which I filter out for you. So a few ocassional out of the office replies is a small price to pay for the information you get for subscribing to this forum.
Cheers,
Nestor Your Friendly Neighborhood SysOp.
==============================Original Headers============================== 12, 13 -- From zaluzec-at-microscopy.com Sat Apr 21 08:25:22 2007 12, 13 -- Received: from [206.69.208.22] (mac22.zaluzec.com [206.69.208.22]) 12, 13 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3LDPJrW012128 12, 13 -- for {microscopy-at-microscopy.com} ; Sat, 21 Apr 2007 08:25:21 -0500 12, 13 -- Mime-Version: 1.0 12, 13 -- Message-Id: {p06240803c24fb999c345-at-[206.69.208.22]} 12, 13 -- In-Reply-To: {200704211136.l3LBaXsH031128-at-ns.microscopy.com} 12, 13 -- References: {200704211136.l3LBaXsH031128-at-ns.microscopy.com} 12, 13 -- Date: Sat, 21 Apr 2007 08:25:18 -0500 12, 13 -- To: microscopy-at-microscopy.com 12, 13 -- From: "Nestor J. Zaluzec" {zaluzec-at-microscopy.com} 12, 13 -- Subject: Out of Office messages & posting comments to the list 12, 13 -- Content-Type: text/plain; charset="us-ascii" ==============================End of - Headers==============================
Software image analysis of TEM film or digital images isn't always easy as it is often difficult for the software to distinguish the objects you wish to count from the rest of the image. However our brains can generally do a far better job in picking out (discriminating) what we want to count or measure. But you have be careful with things like sampling procedure and image quality to make sure you don't bias the results.
Generally, even with modern computers, it is often quicker to count by eye than get software to do this for you. This is particularly so if there are only a few objects (less than 50) in the image. However if you can easily software threshold (detect) the objects you wish to count then the software can count them automatically very quickly. But often the 'detected' thresholded binary overlay will need editing to remove false objects, and this may take far longer than simply counting by eye.
In most cases careful preparation of the specimen and optimising image capture and microscope optics will be essential to produce images are most suited to semi-automatic image analysis software. For example if you wish to count cells under optical microscopy, plate them at lower densities so that they will not overlay each or frequently touch and so can be easily separated and counted by software algorithms. Things like blue DAPI fluorescence staining of the nucleus will enable cell numbers to be counted very quickly semi-automatically, although it will get into trouble if a lot of cells are multi-nuclear or in different focus planes. Often though, even if the software doesn't automatically count the objects as 'accurately' as we can by eye, the biological variation across samples is far higher than these counting errors. Indeed different users will invariably produce different counts for the same samples.
If, as is often the case with TEM and optical phase contrast transmission images, it is very difficult to only threshold the objects of interest, then manual counting is generally the quickest option. Image analysis software can still help here though. Biomedical image analysis programs like MetaMorph (http://www.moleculardevices.com/pages/software/metamorph.html) have applications that allow you to simply click on each object with the mouse to count them. The screen then marks the point with a coloured number (so you can see you have counted it) and it automatically adds up all the objects you have clicked on in the image. You can give different types of objects different coloured markers, so that the count totals go into different bins (up to eight different objects types in one go). Once completed the count data can then be logged to an Excel spreadsheet or noted down.
However MetaMorph image analysis software costs several thousand pounds. On the plus side it can do far more than just manually count (which we are very good at with our object discriminating brain), for example it can also measure length, area and perimeter, track cell movement or estimate co-localisation (which we are far less successful at, even with some sort of reference aid onscreen, e.g. length micrometers or circles of known area).
However you get pretty good PC based image analysis software for free, in the shape of open-source 'ImageJ' image analysis software, see http://rsb.info.nih.gov/ij (or its sister program NIHImage for the apple http://rsb.info.nih.gov/nih-image).
Having many Metamorph licences I have never used ImageJ to count objects, but it has all the standard thresholding (object detection) and image analysis routines (e.g. area, perimeter, length/distance, pixel intensity/brightness). ImageJ can be further expanded by a list of plug-ins written by devoted users (these are just copied to the ImageJ plug-in folder and will then appear in the ImageJ menu, spookily under the word 'Plugins').
ImageJ even has a plug-in that is the exact equivalent of Metamorph's manual count application (See plug-in 'CELL COUNTER'), and it works in a very similar fashion to MetaMorphs version (and for free). You can also simply use a program like Photoshop or even Paint to put a little blob of colour over each object to be counted (destroying the original image so ensure you have a backup). You can then easily threshold and count the blobs with ImageJ (but using plug-in CELL COUNTER is naturally far easier for just counting, colouring in the objects is more useful for things like object area). This Cell Counter software works just like putting a clear plastic sheet over the VDU screen and marking off the objects with a felt pen (and ImageJ counts them for you as well). The software can actually mark the original image so that the counts get scored into it or the count marks marks can be left on an 'overlay' with the original image unchanged (with no record of which objects were counted when the image is next viewed).
These manual count software routines prevent double counting errors and simply forgetting what number you got to (in the old days you used a mechanical one-hand click counter). Like MetaMorph, ImageJ can do an awful lot more image analysis than just manual count. It will always be useful to have an image editing software package like Photoshop at hand as well. Plus a cheap Wacom graphics tablet can be advantageous for some things (like drawing round objects etc..), used in combination with the PC mouse, see www.Wacom.com
If you only have TEM negatives, simply scan them into PC digital form using a cheap flat bed large format (A4) film scanner like the Epson 4990 Photo (£250). ImageJ is happy with standard digital images, e.g. jpg and tif formats.
If you need any more info/help, just ask.
Regards
Keith
--------------------------- Dr Keith J Morris Imaging Facilities Manager Cell Biology Institute of Ophthalmology UCL, London EC1V 9EL
-----Original Message----- X-from: akoorts-at-medic.up.ac.za [mailto:akoorts-at-medic.up.ac.za] Sent: 18 April 2007 14:38 To: keith.morris-at-ucl.ac.uk
This Question was submitted to Ask-A-Microscopist by (akoorts-at-medic.up.ac.za) from http://www.msa.microscopy.org/Ask-A-Microscopist/Ask-A-Microscopist.html on Wednesday, April 18, 2007 at 06:36:15 Remember to consider the Grade/Age of the student when considering the Question --------------------------------------------------------------------------- Please reply to both akoorts-at-medic.up.ac.za as well as to the Microscopy Listserver ---------------------------------------------------------------------------
You are the one to say what you do or don't want. Unless you mean specific brands and/or models of systems.
Buying a new SEM can be a joy and a headache. I can't imagine anything worse than buyer's remorse when what you bought is not what you really needed. To be sure, have a demo for each system you are considering. Bring some typical specimens you need imaged. Also look under the hood (inside the plinth, cabinets, etc.). This will give you an idea of the level of construction technology the tool is based on. You can also find out how well or badly the SE detector works at low vacuum. At up to about 8KX, they tend to work OK.
For low vacuum work, an ESEM is probably what you need. Another option is the Zeiss EVO 50 with LaB6 (rather than W). It has the XY movement you want but not as much Z, but it should be plenty. As far as support is concerned, Zeiss has said that they will support instruments for ten years after end of production. Other companies have their own policy. Find out what it is. The Amray SEMs are easy to fix and in my experience, were quite reliable. The new SEMs are quantum leaps in column technology and electronics, not to mention the extent of computer control.
Modern SEMs are digital capture. This is excellent. Native TIFF is perfect for image processing. Analog (film) output is a hassle since one must scan it to digital anyway to use the image. Most current SEMs do not even include Polaroid/Graphloc backs. At a minimum, the captured image will need brightness and contrast adjustment. Some systems produce indexed color TIFF and these require stripping the color off to make the picture look good. This is easy to do with a Photoshop action. Once converted, additional processing can be applied to enhance the area of interest or add contrast to different layers. This may not be necessary for biological specimens but it is quite useful for metallurgical and semiconductor specimens. In this respect, there is no issue of "honesty." The purpose of the picture is to reveal structure, do failure analysis, evaluate fabrication processing, etc.
Another factor that gets little attention is the design of the column and final lens. For an environmental SEM, this likely does not apply. But the issue is whether the final lens is magnetic immersion or electrostatic. With a magnetic immersion system, it does not like Ferrous specimens. The electrostatic system does not care.
As for EDS, much has changed in the last year. Cryo cooled Si(Li) detectors have problems and are likely to go away. With the advent of new generation Peltier cooled Silicon Drift Detectors (SDD), I suspect that LN2 dewar Si(Li) detector's days are numbered.
The past problem (among others) with SDDs was the change/decrease in resolution as counts increased. Si(Li) does not do this. The current generation of SDDs with the FET integrated onto the detector chip have pretty much eliminated this problem. In addition to the ability to handle very high counts, the SDDs are quiet, small and almost instant-on. Do the research to find the SDD that has optimum characteristics. If LN2 is not a bother for you, then going with a Si(Li) dewar detector is probably significantly cheaper than a new generation SDD. Either way, get a EDS "system." One brand of detector and software.
You should prepare a statement of requirements that lists ALL of the features and limits you need. Take your specimens and the requirements list to demo sites. You will then get a good feel for current generation SEMs and EDS. Then, modify your requirements accordingly. If several company's products work for you, then it is just the bid process to select your system.
Let us know how it goes. Since you have one shot at the well, drink deeply. Hopefully you will not wind up saying "If only I'd ....."
gary g.
At 09:50 AM 4/20/2007, you wrote: } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
==============================Original Headers============================== 14, 20 -- From gary-at-gaugler.com Sat Apr 21 15:15:17 2007 14, 20 -- Received: from qsmtp3.mc.surewest.net (qsmtp.mc.surewest.net [66.60.130.145]) 14, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id l3LKFGZW014009 14, 20 -- for {microscopy-at-microscopy.com} ; Sat, 21 Apr 2007 15:15:16 -0500 14, 20 -- Message-Id: {200704212015.l3LKFGZW014009-at-ns.microscopy.com} 14, 20 -- Received: (qmail 15380 invoked from network); 21 Apr 2007 13:15:16 -0700 14, 20 -- Received: by simscan 1.1.0 ppid: 15377, pid: 15378, t: 0.1677s 14, 20 -- scanners: regex: 1.1.0 attach: 1.1.0 14, 20 -- Received: from unknown (HELO ivan.gaugler.com) (66.60.171.211) 14, 20 -- by qsmtp3 with SMTP; 21 Apr 2007 13:15:16 -0700 14, 20 -- X-Mailer: QUALCOMM Windows Eudora Version 7.1.0.9 14, 20 -- Date: Sat, 21 Apr 2007 13:15:19 -0800 14, 20 -- To: gerard.cox-at-goodrich.com 14, 20 -- From: Gary Gaugler {gary-at-gaugler.com} 14, 20 -- Subject: Re: [Microscopy] New SEM Purchase 14, 20 -- Cc: MSA listserver {microscopy-at-microscopy.com} 14, 20 -- In-Reply-To: {200704201750.l3KHoH2u008736-at-ns.microscopy.com} 14, 20 -- References: {200704201750.l3KHoH2u008736-at-ns.microscopy.com} 14, 20 -- Mime-Version: 1.0 14, 20 -- Content-Type: text/plain; charset=us-ascii; format=flowed; x-avg-checked=avg-ok-13265764 ==============================End of - Headers==============================
My opinion is that less than 40% humidity is ideal.
Ted Dunn The EMscope Company Ltd. Thailand
--- TJJ-at-stowers-institute.org 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 } ---------------------------------------------------------------------------- } } Can anybody give me an optimal % relative humidity } that is recommended } for EM rooms? We recently had some failures with } formvar grids (we } think) due to excessive room humidity when they were } being made. I can } bring in a de-humidifier, but what would be } considered to be optimal? } } Teri Johnson, HT(ASCP)QIHC } Managing Director Histology Facility } Stowers Institute for Medical Research } 1000 E. 50th St. } Kansas City, MO 64110 } } } } ==============================Original } Headers============================== } 4, 20 -- From tjj-at-stowers-institute.org Fri Apr 20 } 16:04:09 2007 } 4, 20 -- Received: from stowers-institute.org } (mail.stowers-institute.org [204.56.6.8]) } 4, 20 -- by ns.microscopy.com } (8.12.11.20060308/8.12.8) with ESMTP id } l3KL48qX000489 } 4, 20 -- for {microscopy-at-microscopy.com} ; Fri, 20 } Apr 2007 16:04:08 -0500 } 4, 20 -- X-MimeOLE: Produced By Microsoft Exchange } V6.5 } 4, 20 -- Content-class: urn:content-classes:message } 4, 20 -- MIME-Version: 1.0 } 4, 20 -- Content-Type: text/plain; } 4, 20 -- charset="us-ascii" } 4, 20 -- Subject: Recommended humidity level } 4, 20 -- Date: Fri, 20 Apr 2007 16:03:58 -0500 } 4, 20 -- Message-ID: } {C28BAF593DC3314E9C0F3A50191C2E7807A3DF2D-at-EXCHKC03.stowers-institute.org} } 4, 20 -- X-MS-Has-Attach: } 4, 20 -- X-MS-TNEF-Correlator: } 4, 20 -- Thread-Topic: Recommended humidity level } 4, 20 -- Thread-Index: } AceDjyLmsE1bPtgoQnSyl90Uh5X67QAADZUQ } 4, 20 -- From: "Johnson, Teri" } {TJJ-at-stowers-institute.org} } 4, 20 -- To: {microscopy-at-microscopy.com} } 4, 20 -- Content-Transfer-Encoding: 8bit } 4, 20 -- X-MIME-Autoconverted: from quoted-printable } to 8bit by ns.microscopy.com id l3KL48qX000489 } ==============================End of - } Headers============================== }
__________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com
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You'll be lucky to find a coolant less viscous than water, particularly one with such low cost and toxicity!
For what its worth, with 5 bar (about 75psi) supply pressure, I get
2.0 litres/min thru the diff pumps 1.0 l/min thru the electronics 0.15 l/min thru the OL ie total around 3.2 l/min.
these are lower flows than the JEOL recommendations of 3.0, 1.5, and 0.5 ie total 5.0 l/min, but the combined discharge water is only 4 deg C warmer than the supply and none of the lines feels particularly warm, so all seems well. My 840 hasn't been flushed out for years, its on my to-do-when-I-have-time list.
Your Iwaki pump probably lacks sufficient grunt for this application, 2.1 metres water head is only about 3psi, which has no chance of shifting enough water through the 840.
Don't attempt to run the scope until you sort this out (buy a bigger pump), the DPs have overtemp switches, and the electronics is protected provided you set the little switch on the fuse panel to 'AUTO', but there is no thermal protection for the OL, and you certainly don't want to burn that one out.
good luck with the project
cheers
rtch
On 20 Apr 2007 at 17:44, 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 } ---------------------------------------------------------------------- } ------ } } Sorry, I wasn't very clear about the chiller before. It's not a name } brand chiller- it's actually a part from a larger system, but I don't } know what that larger system is- I bought it locally as industrial } surplus. Here's what I know about the recirculator system: } } Pump: Iwaki Magnetic drive pump. 12 L/min. Max. head 2.1m. } } There is a heat exchanger unit and all of the condenser piping } necessary to chill the water, and it can divide the output a maximum } of three ways, so I can have the electronics set on a different water } circuit than the diffusion pumps. When I say that the pressure is not } enough to push the water through, I mean that the pump pumps, but very } little water is pushed through, and it is shutting down because of } thermal overload. I know that the JEOL manual specifies 5 L/min. at } 12-36 PSI, but I'm not sure how that translates from "Max head" to } PSI. The pump's flow rate is more than enough to handle it, but it } isn't making it through. That's why I was thinking about lower } viscosity fluids, which should flush through at a higher flow rate and } a lower PSI. }
-- 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============================== 14, 27 -- From r.sims-at-auckland.ac.nz Sun Apr 22 18:12:17 2007 14, 27 -- Received: from mailhost.auckland.ac.nz (larry.its.auckland.ac.nz [130.216.10.122]) 14, 27 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3MNCGTR020307 14, 27 -- for {microscopy-at-microscopy.com} ; Sun, 22 Apr 2007 18:12:17 -0500 14, 27 -- Received: from localhost (localhost.localdomain [127.0.0.1]) 14, 27 -- by mailhost.auckland.ac.nz (Postfix) with ESMTP id DB86B18306; 14, 27 -- Mon, 23 Apr 2007 11:12:14 +1200 (NZST) 14, 27 -- X-Virus-Scanned: by amavisd-new at mailhost.auckland.ac.nz 14, 27 -- Received: from mailhost.auckland.ac.nz ([127.0.0.1]) 14, 27 -- by localhost (larry.its.auckland.ac.nz [127.0.0.1]) (amavisd-new, port 10024) 14, 27 -- with ESMTP id Mo5i87en4KKw; Mon, 23 Apr 2007 11:12:14 +1200 (NZST) 14, 27 -- Received: from rs (r.sims.glg.auckland.ac.nz [130.216.59.18]) 14, 27 -- by mailhost.auckland.ac.nz (Postfix) with ESMTP id BE3FE18301; 14, 27 -- Mon, 23 Apr 2007 11:12:14 +1200 (NZST) 14, 27 -- From: "Ritchie Sims" {r.sims-at-auckland.ac.nz} 14, 27 -- To: kraftpiano-at-gmail.com 14, 27 -- Date: Mon, 23 Apr 2007 11:13:04 +1200 14, 27 -- MIME-Version: 1.0 14, 27 -- Subject: 840 water flows 14, 27 -- CC: microscopy-at-microscopy.com 14, 27 -- Message-ID: {462C94C0.19326.C32006-at-localhost} 14, 27 -- Priority: normal 14, 27 -- In-reply-to: {200704202244.l3KMi2qE028582-at-ns.microscopy.com} 14, 27 -- X-mailer: Pegasus Mail for Windows (4.21c) 14, 27 -- Content-type: text/plain; charset=US-ASCII 14, 27 -- Content-transfer-encoding: 7BIT 14, 27 -- Content-description: Mail message body ==============================End of - Headers==============================
I have probably purchased more electron microscopes than anyone else in the world having acted as a consultant on an average of two or three purchases a year over the past 25 years!
The summary of what I have learned is not to eliminate a microscope until you have seen it in action on your specimens! My other point is not to close your mind on future applications. An aircraft engine company were pushed by me into purchasing a FEG instrument. In the first week they found that their ideas of not going above 20,000X and below 15kV were blown away, the additional information they could see was "mind blowing" and critical.
A third area of my concern is the selection of specimens and the clients attitude during a demonstration. As an exdemonstrator for Hitachi, JEOL and ISI I found that many prospective customers fight a silly battle with the demonstrator. Firstly, one often found that quite often the client had not looked at the specimens they had brought they were from another person in the lab! In other cases the game is for the demonstrator, with no help from the client, is expected to produce more and better information in one day than the client has produced over many years. The problems deepen because the client does not try to inform the demonstrator and help the demonstration to bring out the critical data. After all it should not be a test of the demonstrator but better a test of the microscope, most people are buying the best microscope not the best demonstrator but that is difficult to see in some demonstrations ;)
Time and time again the eventual purchase is not the instrument that we start off believing is the best for the client. I have written presentations on instrument purchase all too long to fit in here but available on our web site in hints and tips or in a past edition of Microscopy Today.
I hope this helps?
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: {gerard.cox-at-goodrich.com} To: {protrain-at-emcourses.com} Sent: Friday, April 20, 2007 6:48 PM
Hi,
According to Bernoulli: Static PSI = height* specific gravity/ 2.31 2.31 feet of water is 1 PSI
Your PSI = 6.8 ft * 1 /2.31 = 2.9 PSI of STATIC HEAD
I had a system in a fan loft three stories higher than the lab. The back pressure was about 15 PSI. So 3 stories is about 34 feet of static head. So 34 feet is to ~15 PSI as 6.8 feet is to X
X = ~3.0 PSI for 2.1 meters of STATIC HEAD.
If the fluid is moving, the pressure will be less.
Loss of Head = V²/(2 * 32.2 feet / sec²) At 1 foot per second, Loss of Head = 1/(2*32.2) = 0.016 PSI or almost zero.
You need more pressure from a different direct drive pump, IMHO.
Flow rate: Your quoted flow rate might be and appears to me to be at no head pressure. Flow will decrease as the head pressure is increased from back pressure.
Thermal load? Most DP use at least 600 watts and usually more with Santovac. So your system must have a cooling capacity of at least 600 watts and you probably need to dump 1500 watts of heat as a minimum due to losses from cooling lines.
FYI Notes: The green color in restricted cooling lines is usually the green compound copper carbonate, (CuCO3) or basic copper carbonate. Other materials and compounds are involved. The scale can be removed with minimal damage to lens tubing, if that's the problem but I don't think that is it in this case.
Maintaining a balance of the chemistry in a chiller requires an understanding of the Langalier index to make or remove scale in-situ. The copper carbonate Ksp value I calculated years ago on paper was eliminated by zealots of records retention. I just recalculated it. The solubility used was 0.28 mg per 100 mls, an average value I found on the Internet for copper carbonate. Ksp CuCO3 = 4.96 * 10^(-14) Ksp CaCO3 = 4.8 x 10^(-9) (Skoog and West) or [0.87 x 10^(-8) in the CRC Handbook] So GREEN copper carbonate will PPT before CaCO3 but it is similar to CaCO3 in that both have limited solubility. The amount of calcium from contamination by dead volumes involved in using city water and improper dead volume flushing, should not increase like copper ions do. The green color and the Ksp value of copper carbonate can be used as a "colorimetric spot test". I used this test. All plastic pipes will not save you from copper. Fittings and pump heads are made of brass.
CaCO3, CuO and carbonates of copper are involved as well as the pH and CO2 gas mixing. The scaling can be tracked by observing the color of semitransparent PP tubing in an Edwards 306A or an equivalent transparent tubing somewhere in your system. If you see a green color forming as a lining in the tubing, then the Ksp [ion concentration product] has been exceeded. You need to remove some copper ions and replace that water with distilled water. Don't remove all the "old" water. Leave some ions behind to mix with the new DI water. This is easy to do in a properly designed system with a chilled water line that can divert contaminated water into a lab sink and that also has an automatic DI water refill on the chiller reservoir.
Disclaimers and simple water tests: Copper ions kill algae. Quaternary ammonium salt surfactants (used in swimming pools) kill algae by breaking open cell walls. Adding ammonium hydroxide to a sample of chiller water containing copper ions causes a deep blue color to form at high pH. Chloride ions cause stainless steel to pit corrode. Most DPs and some tubing fittings are made of SS. Silver nitrate solution forms a white precipitate when added to a chilled water sample with chloride ions in it. Any form of available chlorine like chloramine-T or isocyanurates will be reduced to chloride ions over time. Do you really want cyanuric acid or p-toluene-sulfonamide in your system? Using all plastic pipes and an HVAC heat exchanger system to supply chilling, will not save you from copper ions forming.
Paul
At 05:43 PM 4/20/07 -0500, you wrote: } } } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
==============================Original Headers============================== 16, 27 -- From beaurega-at-westol.com Mon Apr 23 07:51:02 2007 16, 27 -- Received: from smtp-gateway-4.winbeam.com (smtp-gateway-4.winbeam.com [64.84.97.69]) 16, 27 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3NCp2k1011130 16, 27 -- for {microscopy-at-microscopy.com} ; Mon, 23 Apr 2007 07:51:02 -0500 16, 27 -- Received: from mail.winbeam.com (mail.winbeam.com [64.84.96.10]) 16, 27 -- by smtp-gateway-4.winbeam.com (8.13.2/8.12.8) with SMTP id l3NCobSx012895 16, 27 -- for {microscopy-at-microscopy.com} ; Mon, 23 Apr 2007 08:50:38 -0400 16, 27 -- Received: (qmail 15379 invoked by uid 89); 23 Apr 2007 12:50:35 -0000 16, 27 -- Received: from pitts-69-72-117-1.dynamic-dialup.coretel.net (HELO beaurega) (69.72.117.1) 16, 27 -- by mail.winbeam.com with SMTP; 23 Apr 2007 12:50:35 -0000 16, 27 -- Message-Id: {3.0.6.32.20070422175711.007bb100-at-pop3.norton.antivirus} 16, 27 -- X-Sender: beaurega/mail.westol.com-at-pop3.norton.antivirus (Unverified) 16, 27 -- X-Mailer: QUALCOMM Windows Eudora Light Version 3.0.6 (32) 16, 27 -- Date: Sun, 22 Apr 2007 17:57:11 -0400 16, 27 -- To: kraftpiano-at-gmail.com, microscopy-at-microscopy.com 16, 27 -- From: Beaurega {beaurega-at-westol.com} 16, 27 -- Subject: Re: [Microscopy] SEM: Replacing diffusion pump water cycle 16, 27 -- with coolant. 16, 27 -- In-Reply-To: {200704202243.l3KMhatn027698-at-ns.microscopy.com} 16, 27 -- Mime-Version: 1.0 16, 27 -- Content-Type: text/plain; charset="iso-8859-1" 16, 27 -- Content-Transfer-Encoding: 8bit 16, 27 -- X-smtp-gateway-4-MailScanner-Information: smtp-gateway-4 - Please contact Technical Support for more information 16, 27 -- X-smtp-gateway-4-MailScanner: Found to be clean smtp-gateway-4 (courtesy of MailScanner) 16, 27 -- X-smtp-gateway-4-MailScanner-SpamCheck: not spam (whitelisted), 16, 27 -- SpamAssassin (not cached, timed out) 16, 27 -- X-smtp-gateway-4-MailScanner-From: beaurega-at-westol.com ==============================End of - Headers==============================
Our ASI stage with MS-2000 controller has not worked properly with IPLab for years. Back with IPLab versions 3.65 and earlier we wrote our own scripts to get around the problem, but with the upgrade to 4.0.# the ASI stage completely does not work with the IPLab software.
For *more*than*a*year* we have been asking Scanalytics / BD BioSciences to fix this problem and even though we have had visits on site, the problem is not solved. We invested in multiple license upgrades, and two of the systems simply don't work.
Does anybody know how to operate the ASI stage with a PC running IPLab 4.0.#?
At this point we are desperate to have our two inverted microscopes with mechanical stages work properly and unless this email to a public listserv acts as a motivator, I am ready to give up on IPLab as a software solution for our light microscopy needs and BD Biosciences as truly responsive to customers.
-Michael Cammer ____________________________________________________________________________ Michael Cammer Analytical Imaging Facility Albert Einstein Coll. of Med. URL: http://www.aecom.yu.edu/aif/
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Thanks for all your replies. Just to clarify, this will be in a sample prep area only, the scope is in a different room. The ventilation in this room is "weird" (the best way I can describe it), most of the time it's not an issue. On occasion though, you can walk in to the room from the hallway and the atmospheric conditions are totally different.
I'm thinking between 30 and 40% sounds about right.
==============================Original Headers============================== 3, 20 -- From tjj-at-stowers-institute.org Mon Apr 23 09:56:05 2007 3, 20 -- Received: from stowers-institute.org (mail.stowers-institute.org [204.56.6.8]) 3, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3NEu4nd004651 3, 20 -- for {microscopy-at-microscopy.com} ; Mon, 23 Apr 2007 09:56:05 -0500 3, 20 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 3, 20 -- Content-class: urn:content-classes:message 3, 20 -- MIME-Version: 1.0 3, 20 -- Content-Type: text/plain; 3, 20 -- charset="us-ascii" 3, 20 -- Subject: FW: Recommended humidity level 3, 20 -- Date: Mon, 23 Apr 2007 09:55:48 -0500 3, 20 -- Message-ID: {C28BAF593DC3314E9C0F3A50191C2E7807A3DF30-at-EXCHKC03.stowers-institute.org} 3, 20 -- X-MS-Has-Attach: 3, 20 -- X-MS-TNEF-Correlator: 3, 20 -- Thread-Topic: Recommended humidity level 3, 20 -- Thread-Index: AceDjyLmsE1bPtgoQnSyl90Uh5X67QAADZUQAIni5RA= 3, 20 -- From: "Johnson, Teri" {TJJ-at-stowers-institute.org} 3, 20 -- To: {microscopy-at-microscopy.com} 3, 20 -- Content-Transfer-Encoding: 8bit 3, 20 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l3NEu4nd004651 ==============================End of - Headers==============================
On Apr 20, 2007, at 6:49 PM, gary-at-gaugler.com wrote:
} The reason NOT to post to the list is that you get } volumes of "Out Of Office" messages. This is the } same as when you make a new posting. Most of the } defunct messages are trapped by Eudora but quite } a few still get though....sigh. } } Some do not have this subject but say that they } are gone from xx to yy. } Dear Gary, Is it possible to have all OOO messages have the subject line be "Out Of Office"? If so, Nestor could request it, but if the company that one works for has a standard OOO heading, that may be impractical. 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 Mon Apr 23 11:35:09 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 l3NGZ94x018840 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Mon, 23 Apr 2007 11:35:09 -0500 4, 22 -- Received: from fire-dog.caltech.edu (fire-dog [192.168.1.4]) 4, 22 -- by water-ox-postvirus (Postfix) with ESMTP id 4B6AF2F51F 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Mon, 23 Apr 2007 09:35:02 -0700 (PDT) 4, 22 -- Received: from [192.168.159.233] (jpix-01.caltech.edu [131.215.2.133]) 4, 22 -- by earth-ox.its.caltech.edu (Postfix) with ESMTP id C778D3709B 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Mon, 23 Apr 2007 09:35:00 -0700 (PDT) 4, 22 -- Mime-Version: 1.0 (Apple Message framework v624) 4, 22 -- In-Reply-To: {200704210149.l3L1nrNR031977-at-ns.microscopy.com} 4, 22 -- References: {200704210149.l3L1nrNR031977-at-ns.microscopy.com} 4, 22 -- Content-Type: text/plain; charset=US-ASCII; format=flowed 4, 22 -- Message-Id: {f47cb397bd5c315ec4c834c9aa068665-at-caltech.edu} 4, 22 -- Content-Transfer-Encoding: 7bit 4, 22 -- From: Bill Tivol {tivol-at-caltech.edu} 4, 22 -- Subject: Re: [Microscopy] Re: 2nd for posting to the list 4, 22 -- Date: Mon, 23 Apr 2007 09:46:01 -0700 4, 22 -- To: microscopy-at-msa.microscopy.com 4, 22 -- X-Mailer: Apple Mail (2.624) 4, 22 -- X-Spam-Scanned: at Caltech-ITS on fire-dog by amavisd-2.3.3 ==============================End of - Headers==============================
We are pleased to announce that the Oxford Brookes University Masters course in Bioimaging will begin in Sept. 2007. There is still room for students and information about the course can be found at the following link:
We will run this as a modular one year full time course intended to teach microscopy (light / confocal / em) and associated laboratory skills.
If you know of students or professionals that might be interested, please can you pass this information to them. Please contact me if you need more detailed information or if you would like a pdf that provides details of the course.
Sincerely, John. --
********************************* C. John Runions, Ph.D. School of Life Sciences Oxford Brookes University Oxford, UK OX3 0BP
This is the first announcement for students who will attend the Microscopy and Microanalysis Meeting in Ft. Lauderdale, Fl, August 5-9, 2007 and would like to apply for a Student Bursary to help defray meeting costs. The complete description of the Bursary is listed below, including the registration process.
The contact persons this year who will coordinate the student volunteers are;
Bill Monroe monroe-at-emcenter.msstate.edu Amanda Lawrence lawrence-at-emcenter.msstate.edu Mike Miller millem1-at-auburn.edu
STUDENT BURSARIES
The Microscopy Society of America values student members and recognizes that they are the future of both the society and the field of microscopy in general. MSA is therefore pleased to offer student bur-saries of $200 to registered students. The most important purpose of these bursaries is to encourage students to attend the annual MSA/MAS Microscopy and Microanalysis meeting, where these young sci-entists can meet and interact with the established microscopy community as well as assisting with the meeting.
Each bursary recipient will be expected to work for a minimum of 20 hours during the meeting and/or at the pre-meeting weekend events. A student may work up to an additional 20 hours, for a total of 40 hours. These extra hours will add to the bursary total at a rate of $10 per hour. The maximum bursary will therefore be $400. The duties will involve, but are not necessarily limited to, providing support in symposia (helping with audio-visual needs, maintaining an attendance count, and helping speakers set up for their presentation), staffing the MSA Megabooth, and monitoring use of the Internet Cafe. Applicants for the bursaries must be members of MSA or MAS, and enrolled as students at a recognized educational institution. All MSA or MAS student members are eligible for bursaries, including those who are recipients of MSA Presidential Scholars Awards and MAS Distinguished Scholar Awards. Eligible students may apply for bursaries when registering for the conference on the conference website, or at on-site registration. Bursaries are limited and early application is encouraged. How it works:
The registration form for the meeting will have a check box indicating that the applicant is a registered student and is requesting a bursary. The check box will have a note beside it reminding the applicant that the bursary requires them to work at the meeting. Students who have applied for bursaries will receive letters from MSA explaining the conditions that they need to satisfy in order to receive the bursaries. The tasks at the meeting will be allocated by the Student Worker Organizers Sub-Committee of the Education Committee. When students pick up their registration materials at the meeting, they will receive assignment forms indicating the specific tasks they are to perform, and the person(s) they need to contact in order to carry out those tasks. Each stu-dent's assignment forms must be signed by all of the contact persons listed, indicating that all assigned tasks have been performed. Upon completion of assignments and submission of the signed forms, the bursary check will be issued by the appropriate representative at the registration desk.
Should you have questions concerning the process, please contact:
Bill Monroe monroe-at-emcenter.msstate.edu
-- Bill Monroe Electron Microscope Center 103 Clay Lyle Entomology Building Mississippi State University Mississippi State, MS 39762 (662)-325-3019 Work (662)-323-5246 Home (662)-325-0246 Fax
==============================Original Headers============================== 16, 14 -- From monroe-at-emcenter.msstate.edu Mon Apr 23 15:02:18 2007 16, 14 -- Received: from Ra.MsState.Edu (Ra.MsState.Edu [130.18.80.10]) 16, 14 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3NK2HBL014594 16, 14 -- for {Microscopy-at-microscopy.com} ; Mon, 23 Apr 2007 15:02:17 -0500 16, 14 -- Received: from [130.18.130.108] (ws108-130.clay-lyle.dynamic.msstate.edu [130.18.130.108]); 16, 14 -- by Ra.MsState.Edu (8.13.8/8.12.8/ra_1.2) with SMTP; 16, 14 -- id l3NK2G3k026969 for {Microscopy-at-microscopy.com} ; Mon, 23 Apr 2007 15:02:16 -0500 (CDT) 16, 14 -- Mime-Version: 1.0 16, 14 -- Message-Id: {a06230907c252c11180bb-at-[130.18.130.108]} 16, 14 -- Date: Mon, 23 Apr 2007 15:02:15 -0500 16, 14 -- To: Microscopy-at-microscopy.com 16, 14 -- From: "William A. Monroe" {monroe-at-emcenter.msstate.edu} 16, 14 -- Subject: MSA 2007 Meeting: Request for Student Volunteers (Bursaries) 16, 14 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" ==============================End of - Headers==============================
Recently I started working on visualization of bacteria on various biocidal surfaces. I have been soliciting advices regarding samples prep with excellent success for which I should thank Servers. I have new question regarding implementation of cationic dyes for SEM bacterial visualization. Considering that I am primarily looking for changes (sometimes very subtle) in bacterial appearance due to the biocide action I am concern that some of the changes could be introduced by fixation/processing artifacts. Will addition of cationic dye to glutaraldehyde/osmium tetroxide protocol help me to preserve fine structures? Which dye would be best and in which concentration? Would it make difference for gram-positive as much as gram ?negative strains? 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============================== 5, 22 -- From amich-at-ufl.edu Mon Apr 23 15:57:57 2007 5, 22 -- Received: from smtp.ufl.edu (smtp01.osg.ufl.edu [128.227.74.149]) 5, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3NKvvco027141 5, 22 -- for {microscopy-at-microscopy.com} ; Mon, 23 Apr 2007 15:57:57 -0500 5, 22 -- Received: from osgjas01.cns.ufl.edu (osgjas01.cns.ufl.edu [128.227.74.131]) 5, 22 -- by smtp.ufl.edu (8.13.7/8.13.7/2.5.9) with ESMTP id l3NKvsII831724 5, 22 -- for {microscopy-at-microscopy.com} ; Mon, 23 Apr 2007 16:57:55 -0400 5, 22 -- Message-ID: {1674682827.339521177361874691.JavaMail.osg-at-osgjas01.cns.ufl.edu} 5, 22 -- Date: Mon, 23 Apr 2007 16:57:54 -0400 (EDT) 5, 22 -- From: "MIKHAYLOVA,ALBINA" {amich-at-ufl.edu} 5, 22 -- To: microscopy-at-microscopy.com 5, 22 -- Subject: cationic dyes for SEM bacterial visualization 5, 22 -- MIME-Version: 1.0 5, 22 -- Content-Type: text/plain; format=flowed; charset=us-ascii 5, 22 -- Content-Transfer-Encoding: 7bit 5, 22 -- X-Mailer: GatorMail WebMail (http://GatorMail.sf.net/) 5, 22 -- X-Originating-IP: 72.155.91.203 [72.155.91.203] 5, 22 -- X-Greylist: Sender IP whitelisted, not delayed by milter-greylist-3.0rc3 (smtp.ufl.edu [128.227.74.56]); Mon, 23 Apr 2007 16:57:55 -0400 (EDT) 5, 22 -- X-Spam-Status: hits=-1.44, required=5, tests=ALL_TRUSTED 5, 22 -- X-UFL-Spam-Status: hits=-1.44, required=5, tests=ALL_TRUSTED 5, 22 -- X-Scanned-By: CNS Open Systems Group (http://open-systems.ufl.edu/services/smtp-relay/) 5, 22 -- X-UFL-Scanned-By: CNS Open Systems Group (http://open-systems.ufl.edu/services/smtp-relay/) ==============================End of - Headers==============================
Both ruthenium red and alcian blue can be used. In addition cetyl pyridimium chloride is also a choice for preserving negatively charged mucopolysaccharides that are on the surface of some cells. The tendency is to overly decorate these surface coats and obscure real detail. These reagents do nothing to preserve the actual cell or the cells wall. I also think that SEM is not the tool for seeing any subtle effects. Work I had done some years ago required thin section TEM and even then only catastrophic changes were evident i.e. lysis or cytoplasmic partitioning. Changes in membrane architecture might require freeze fracture EM. To minimize fixation artefacts for SEM you might try rapid freezing followed by freeze drying; freeze substitution fixation followed by critical point drying or cryo-SEM of fully hydrated cells. In all three of these techniques very rapid freezing is required in order to prevent ice crystal formation that would cause artefact in and of itself. There is a book in the UF library by Aldrich and Taylor that discusses in more detail some of the things that I have mentioned. I think the title is "Ultrastructure Techniques for Microorganisms". It is old, but many of the techniques are still in use today. I believe that I left my personal copy at the ICBR EM Core Lab, when I retired.
amich-at-ufl.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 Servers, } } Recently I started working on visualization of bacteria on various } biocidal surfaces. I have been soliciting advices regarding } samples prep with excellent success for which I should thank } Servers. } I have new question regarding implementation of cationic dyes for } SEM bacterial visualization. Considering that I am primarily } looking for changes (sometimes very subtle) in bacterial } appearance due to the biocide action I am concern that some of the } changes could be introduced by fixation/processing artifacts. Will } addition of cationic dye to glutaraldehyde/osmium tetroxide } protocol help me to preserve fine structures? Which dye would be } best and in which concentration? Would it make difference for } gram-positive as much as gram ?negative strains? } 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============================== } 5, 22 -- From amich-at-ufl.edu Mon Apr 23 15:57:57 2007 } 5, 22 -- Received: from smtp.ufl.edu (smtp01.osg.ufl.edu [128.227.74.149]) } 5, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3NKvvco027141 } 5, 22 -- for {microscopy-at-microscopy.com} ; Mon, 23 Apr 2007 15:57:57 -0500 } 5, 22 -- Received: from osgjas01.cns.ufl.edu (osgjas01.cns.ufl.edu [128.227.74.131]) } 5, 22 -- by smtp.ufl.edu (8.13.7/8.13.7/2.5.9) with ESMTP id l3NKvsII831724 } 5, 22 -- for {microscopy-at-microscopy.com} ; Mon, 23 Apr 2007 16:57:55 -0400 } 5, 22 -- Message-ID: {1674682827.339521177361874691.JavaMail.osg-at-osgjas01.cns.ufl.edu} } 5, 22 -- Date: Mon, 23 Apr 2007 16:57:54 -0400 (EDT) } 5, 22 -- From: "MIKHAYLOVA,ALBINA" {amich-at-ufl.edu} } 5, 22 -- To: microscopy-at-microscopy.com } 5, 22 -- Subject: cationic dyes for SEM bacterial visualization } 5, 22 -- MIME-Version: 1.0 } 5, 22 -- Content-Type: text/plain; format=flowed; charset=us-ascii } 5, 22 -- Content-Transfer-Encoding: 7bit } 5, 22 -- X-Mailer: GatorMail WebMail (http://GatorMail.sf.net/) } 5, 22 -- X-Originating-IP: 72.155.91.203 [72.155.91.203] } 5, 22 -- X-Greylist: Sender IP whitelisted, not delayed by milter-greylist-3.0rc3 (smtp.ufl.edu [128.227.74.56]); Mon, 23 Apr 2007 16:57:55 -0400 (EDT) } 5, 22 -- X-Spam-Status: hits=-1.44, required=5, tests=ALL_TRUSTED } 5, 22 -- X-UFL-Spam-Status: hits=-1.44, required=5, tests=ALL_TRUSTED } 5, 22 -- X-Scanned-By: CNS Open Systems Group (http://open-systems.ufl.edu/services/smtp-relay/) } 5, 22 -- X-UFL-Scanned-By: CNS Open Systems Group (http://open-systems.ufl.edu/services/smtp-relay/) } ==============================End of - Headers============================== }
-- Greg Erdos University of Florida, Retired Micanopy, Florida
==============================Original Headers============================== 3, 26 -- From gwe-at-ufl.edu Mon Apr 23 16:50:15 2007 3, 26 -- Received: from smtp.ufl.edu (smtp02.osg.ufl.edu [128.227.74.165]) 3, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3NLoExg007029 3, 26 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 23 Apr 2007 16:50:15 -0500 3, 26 -- Received: from [10.228.0.103] (ssrb-vpn1-0-103.vpn.ufl.edu [10.228.0.103]) 3, 26 -- (authenticated bits=0) 3, 26 -- by smtp.ufl.edu (8.13.7/8.13.7/2.5.9) with ESMTP id l3NLo68G5107754; 3, 26 -- Mon, 23 Apr 2007 17:50:07 -0400 3, 26 -- Message-ID: {462D2A0E.3000908-at-ufl.edu} 3, 26 -- Date: Mon, 23 Apr 2007 17:50:06 -0400 3, 26 -- From: Greg Erdos {gwe-at-ufl.edu} 3, 26 -- Reply-To: gwe-at-ufl.edu 3, 26 -- User-Agent: Thunderbird 1.5.0.10 (Windows/20070221) 3, 26 -- MIME-Version: 1.0 3, 26 -- To: amich-at-ufl.edu, 3, 26 -- "Microscopy-at-sparc5.microscopy.com" {Microscopy-at-MSA.Microscopy.Com} 3, 26 -- Subject: Re: [Microscopy] cationic dyes for SEM bacterial visualization 3, 26 -- References: {200704232058.l3NKwTDc028249-at-ns.microscopy.com} 3, 26 -- In-Reply-To: {200704232058.l3NKwTDc028249-at-ns.microscopy.com} 3, 26 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 3, 26 -- Content-Transfer-Encoding: 7bit 3, 26 -- X-Greylist: Sender succeeded SMTP AUTH authentication, not delayed by milter-greylist-3.0rc3 (smtp.ufl.edu [128.227.74.56]); Mon, 23 Apr 2007 17:50:07 -0400 (EDT) 3, 26 -- X-Spam-Status: hits=-1.44, required=5, tests=ALL_TRUSTED 3, 26 -- X-UFL-Spam-Status: hits=-1.44, required=5, tests=ALL_TRUSTED 3, 26 -- X-Scanned-By: CNS Open Systems Group (http://open-systems.ufl.edu/services/smtp-relay/) 3, 26 -- X-UFL-Scanned-By: CNS Open Systems Group (http://open-systems.ufl.edu/services/smtp-relay/) ==============================End of - Headers==============================
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Question: I would like to obtain and install an external scan interface for my Zeiss DSM-950. I believe that any ESI from the DSM series scopes will work. Does anybody know of a retired machine with this card in it? Is anybody out there knowledgeable in installing said interface.
Can anybody direct me to software for producing anaglyphs from 3d data I have acquired using my LSCM? Preferably software which is free or available as a trial.
thanks
----------------------- Adrian A. Evans
==============================Original Headers============================== 6, 22 -- From A.A.Evans-at-Bradford.ac.uk Tue Apr 24 08:01:00 2007 6, 22 -- Received: from hydrogen.cen.brad.ac.uk (hydrogen.cen.brad.ac.uk [143.53.238.3]) 6, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3OD0xxi016640 6, 22 -- for {Microscopy-at-Microscopy.Com} ; Tue, 24 Apr 2007 08:00:59 -0500 6, 22 -- Received: from radon.cen.brad.ac.uk (radon.cen.brad.ac.uk [143.53.238.18]) 6, 22 -- by hydrogen.cen.brad.ac.uk (8.13.6/8.13.4) with ESMTP id l3OD0vgZ028457 6, 22 -- for {Microscopy-at-Microscopy.Com} ; Tue, 24 Apr 2007 14:00:59 +0100 (BST) 6, 22 -- Received: from microwear (microwear.arch.brad.ac.uk [143.53.48.209]) 6, 22 -- by radon.cen.brad.ac.uk (8.13.6/8.13.4) with ESMTP id l3OCweV1009879 6, 22 -- for {Microscopy-at-Microscopy.Com} ; Tue, 24 Apr 2007 13:58:41 +0100 (BST) 6, 22 -- From: "Adrian A. Evans" {A.A.Evans-at-Bradford.ac.uk} 6, 22 -- To: {Microscopy-at-Microscopy.Com} 6, 22 -- Subject: LSCM data and anaglyphs 6, 22 -- Date: Tue, 24 Apr 2007 13:58:35 +0100 6, 22 -- Message-ID: {000501c78670$4551db00$d130358f-at-microwear} 6, 22 -- MIME-Version: 1.0 6, 22 -- Content-Type: text/plain; 6, 22 -- charset="us-ascii" 6, 22 -- Content-Transfer-Encoding: 7bit 6, 22 -- X-Mailer: Microsoft Office Outlook 11 6, 22 -- Thread-Index: AceGcESp1PoPylf4Tkeo9N+dreDMHg== 6, 22 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.3028 ==============================End of - Headers==============================
Our imaging facility is considering acquiring a spinning disk confocal to compliment our 2 laser scanning instruments (Radiance 2000, Zeiss 510 NLO META). The leading configurations are the Yokogawa and Olympus DSU units with an EMCCD. I would appreciate insight on the following issues: Are service contracts on the spinning disk unit or EMCCD essential (I have them on my LSMs)? For those of you running core facilities, does the spinning disk siphon a significant proportion of users off the LSMs or do you find its use restricted to live cell/fast acquisition studies? And finally, how much does going with laser excitation really improve things over the use of a mercury arc or metal halide illumination source? Thanks for any comments you have on this. Tom
Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 2 Tucker Hall University of Missouri Columbia, MO 65211-7400
Don't drop the humidity too low or static charge becomes at least annoying and at worst a problem.
I would say 35% at the lowest.
Ted Dunn The EMscope Company Ltd. Thailand --- TJJ-at-stowers-institute.org wrote:
} ---------------------- } } Thanks for all your replies. Just to clarify, this } will be in a sample } prep area only, the scope is in a different room. } The ventilation in } this room is "weird" (the best way I can describe } it), most of the time } it's not an issue. On occasion though, you can walk } in to the room from } the hallway and the atmospheric conditions are } totally different. } } I'm thinking between 30 and 40% sounds about right. } } } ==============================Original } Headers============================== } 3, 20 -- From tjj-at-stowers-institute.org Mon Apr 23 } 09:56:05 2007 } 3, 20 -- Received: from stowers-institute.org } (mail.stowers-institute.org [204.56.6.8]) } 3, 20 -- by ns.microscopy.com } (8.12.11.20060308/8.12.8) with ESMTP id } l3NEu4nd004651 } 3, 20 -- for {microscopy-at-microscopy.com} ; Mon, 23 } Apr 2007 09:56:05 -0500 } 3, 20 -- X-MimeOLE: Produced By Microsoft Exchange } V6.5 } 3, 20 -- Content-class: urn:content-classes:message } 3, 20 -- MIME-Version: 1.0 } 3, 20 -- Content-Type: text/plain; } 3, 20 -- charset="us-ascii" } 3, 20 -- Subject: FW: Recommended humidity level } 3, 20 -- Date: Mon, 23 Apr 2007 09:55:48 -0500 } 3, 20 -- Message-ID: } {C28BAF593DC3314E9C0F3A50191C2E7807A3DF30-at-EXCHKC03.stowers-institute.org} } 3, 20 -- X-MS-Has-Attach: } 3, 20 -- X-MS-TNEF-Correlator: } 3, 20 -- Thread-Topic: Recommended humidity level } 3, 20 -- Thread-Index: } AceDjyLmsE1bPtgoQnSyl90Uh5X67QAADZUQAIni5RA= } 3, 20 -- From: "Johnson, Teri" } {TJJ-at-stowers-institute.org} } 3, 20 -- To: {microscopy-at-microscopy.com} } 3, 20 -- Content-Transfer-Encoding: 8bit } 3, 20 -- X-MIME-Autoconverted: from quoted-printable } to 8bit by ns.microscopy.com id l3NEu4nd004651 } ==============================End of - } Headers============================== }
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I need an imaging standard that would allow me to check the magnification and distortions present for our SEM. A good example is the set of SIRA gratings, but I thought I'd put it to the list for finding the most versatile reference standard relative to cost. My needs are (1) to check magnification in x & y for high and low mag, (2) to check orthogonality at high and low mag, and (3) to check for barrel, pin-cushioning, or trapazoidal distortions, at high and low mag ... Where high and low mag are } 2000x and {500x.
Tia & cheerios from "the rock" :o) michael shaffer
SEM/MLA Research Coordinator {http://www.mun.ca/creait/maf/} Inco Innovation Centre Memorial University St. John's, Newfoundland
==============================Original Headers============================== 5, 18 -- From michael-at-shaffer.net Wed Apr 25 08:10:15 2007 5, 18 -- Received: from n034.sc1.he.tucows.com (smtpout1476.sc1.he.tucows.com [64.97.157.176]) 5, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3PDAFp3016132 5, 18 -- for {Microscopy-at-microscopy.com} ; Wed, 25 Apr 2007 08:10:15 -0500 5, 18 -- Received: from roamingwolf (134.153.130.141) by n034.sc1.he.tucows.com (7.2.069.1) (authenticated as michael-at-shaffer.net) 5, 18 -- id 4626955E00122F30 for Microscopy-at-microscopy.com; Wed, 25 Apr 2007 13:10:10 +0000 5, 18 -- From: "michael shaffer" {michael-at-shaffer.net} 5, 18 -- To: "MSA Microscopy list" {Microscopy-at-microscopy.com} 5, 18 -- Subject: SEM standards for imaging 5, 18 -- Date: Wed, 25 Apr 2007 10:40:08 -0230 5, 18 -- Message-ID: {000201c7873b$0d874ea0$8d829986-at-CREAIT.MUN.CA} 5, 18 -- MIME-Version: 1.0 5, 18 -- Content-Type: text/plain; 5, 18 -- charset="us-ascii" 5, 18 -- Content-Transfer-Encoding: 7bit 5, 18 -- X-Mailer: Microsoft Office Outlook 11 5, 18 -- X-MIMEOLE: Produced By Microsoft MimeOLE V6.00.2900.3028 5, 18 -- Thread-index: AceHOwxUAS+JgyTXQ/2d8DZqc+Y41A== ==============================End of - Headers==============================
I am using the Geller (http://www.gellermicro.com/mag_standards/mrs.html) MRS-3. It sure is not inexpensive, but should do what you want. ...There are newer versions as well. Be advised the standards are traceable to NPL rather than directly to NIST. Joe Geller should have a web link available indicating an agreement between NIST and NPL (UK) to accept each others certifications. This satisfied my QA department. No connection with Geller other than a satisfied customer.
Woody White BWXT Services
-----Original Message----- X-from: michael-at-shaffer.net [mailto:michael-at-shaffer.net] Sent: Wednesday, April 25, 2007 9:11 AM To: White, Woody N.
Hello MSA'rs :o)
I need an imaging standard that would allow me to check the magnification and distortions present for our SEM. A good example is the set of SIRA gratings, but I thought I'd put it to the list for finding the most versatile reference standard relative to cost. My needs are (1) to check magnification in x & y for high and low mag, (2) to check orthogonality at high and low mag, and (3) to check for barrel, pin-cushioning, or trapazoidal distortions, at high and low mag ... Where high and low mag are } 2000x and {500x.
Tia & cheerios from "the rock" :o) michael shaffer
SEM/MLA Research Coordinator {http://www.mun.ca/creait/maf/} Inco Innovation Centre Memorial University St. John's, Newfoundland
==============================Original Headers============================== 5, 18 -- From michael-at-shaffer.net Wed Apr 25 08:10:15 2007 5, 18 -- Received: from n034.sc1.he.tucows.com (smtpout1476.sc1.he.tucows.com [64.97.157.176]) 5, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3PDAFp3016132 5, 18 -- for {Microscopy-at-microscopy.com} ; Wed, 25 Apr 2007 08:10:15 -0500 5, 18 -- Received: from roamingwolf (134.153.130.141) by n034.sc1.he.tucows.com (7.2.069.1) (authenticated as michael-at-shaffer.net) 5, 18 -- id 4626955E00122F30 for Microscopy-at-microscopy.com; Wed, 25 Apr 2007 13:10:10 +0000 5, 18 -- From: "michael shaffer" {michael-at-shaffer.net} 5, 18 -- To: "MSA Microscopy list" {Microscopy-at-microscopy.com} 5, 18 -- Subject: SEM standards for imaging 5, 18 -- Date: Wed, 25 Apr 2007 10:40:08 -0230 5, 18 -- Message-ID: {000201c7873b$0d874ea0$8d829986-at-CREAIT.MUN.CA} 5, 18 -- MIME-Version: 1.0 5, 18 -- Content-Type: text/plain; 5, 18 -- charset="us-ascii" 5, 18 -- Content-Transfer-Encoding: 7bit 5, 18 -- X-Mailer: Microsoft Office Outlook 11 5, 18 -- X-MIMEOLE: Produced By Microsoft MimeOLE V6.00.2900.3028 5, 18 -- Thread-index: AceHOwxUAS+JgyTXQ/2d8DZqc+Y41A== ==============================End of - Headers==============================
==============================Original Headers============================== 15, 28 -- From nwwhite-at-bwxt.com Wed Apr 25 08:32:50 2007 15, 28 -- Received: from bwxt.com (lynsmtp1.bwxt.com [131.184.146.8]) 15, 28 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3PDWn2B028168 15, 28 -- for {microscopy-at-msa.microscopy.com} ; Wed, 25 Apr 2007 08:32:50 -0500 15, 28 -- Received: from ([131.184.13.224]) 15, 28 -- by lynsmtp1.bwxt.com with ESMTP id 5202488.4715262; 15, 28 -- Wed, 25 Apr 2007 09:32:38 -0400 15, 28 -- Received: from BWXSPO01.BWXS.BWXTECH.NET ([131.184.13.31]) by bwxslynbh01.BWXS.BWXTECH.NET with Microsoft SMTPSVC(6.0.3790.1830); 15, 28 -- Wed, 25 Apr 2007 09:32:38 -0400 15, 28 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 15, 28 -- Content-class: urn:content-classes:message 15, 28 -- MIME-Version: 1.0 15, 28 -- Content-Type: text/plain; 15, 28 -- charset="us-ascii" 15, 28 -- Subject: RE: [Microscopy] SEM standards for imaging 15, 28 -- Date: Wed, 25 Apr 2007 09:32:37 -0400 15, 28 -- Message-ID: {360DDAABED436B49BA397357CCEE8BB7F878FA-at-BWXSPO01.BWXS.BWXTECH.NET} 15, 28 -- In-Reply-To: {200704251311.l3PDB6U1016923-at-ns.microscopy.com} 15, 28 -- X-MS-Has-Attach: 15, 28 -- X-MS-TNEF-Correlator: 15, 28 -- Thread-Topic: [Microscopy] SEM standards for imaging 15, 28 -- Thread-Index: AceHO19r1vgpjK6PQ7SsmYkHtdnAFAAAfSkg 15, 28 -- From: "White, Woody N." {NWWhite-at-bwxt.com} 15, 28 -- To: {michael-at-shaffer.net} , 15, 28 -- "MicroscopyListServer" {Microscopy-at-msa.microscopy.com} 15, 28 -- X-OriginalArrivalTime: 25 Apr 2007 13:32:38.0130 (UTC) FILETIME=[30E3B520:01C7873E] 15, 28 -- Content-Transfer-Encoding: 8bit 15, 28 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l3PDWn2B028168 ==============================End of - Headers==============================
O.k., folks Lets see if the Microscopy and Microanalysis community can come an agreement.
Traditionally the term "Microprobe" has been used to refer to a dedicated wave length dispersive x-ray microanalysis microscope/instrument (XWDS or WDS). Is this still a valid use for the the term "Microprobe"? Or is it antiquated?
. . . because it comes into conflict a "newer" use of the word "microprobe" as refering to a "small" physical probing device, typically mechanical, for manipulating or testing small items. Would these probing-manipuating-testing devices be better called "Nanoprobes" (even thought they may actually handle items / objects in the micrometer to 100's-micrometer range).
Context will NOT sufice to distinguish between the two as microscopists now deal with both.
Anyone care to share comments or opinions ?
Richard E. Edelmann, Ph.D. EXPO Editor, Microscopy and Microanalysis Supplement Electron Microscopy Facility Director 364 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu http://www.emf.muohio.edu
==============================Original Headers============================== 8, 22 -- From edelmare-at-muohio.edu Wed Apr 25 15:42:26 2007 8, 22 -- Received: from mulnx11.mcs.muohio.edu (mulnx11.mcs.muohio.edu [134.53.6.66]) 8, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3PKgPg0028602 8, 22 -- for {microscopy-at-Microscopy.com} ; Wed, 25 Apr 2007 15:42:26 -0500 8, 22 -- Received: from mulnx24.mcs.muohio.edu (mulnx24.mcs.muohio.edu [134.53.6.11]) 8, 22 -- by mulnx11.mcs.muohio.edu (Switch-3.1.8/Switch-3.1.7) with ESMTP id l3PKgPSu007140 8, 22 -- for {microscopy-at-Microscopy.com} ; Wed, 25 Apr 2007 16:42:25 -0400 8, 22 -- Received: from [192.168.1.23] ([134.53.14.105]) 8, 22 -- by mulnx24.mcs.muohio.edu (Switch-3.1.8/Switch-3.1.7) with ESMTP id l3PKgOST020652 8, 22 -- for {microscopy-at-Microscopy.com} ; Wed, 25 Apr 2007 16:42:24 -0400 8, 22 -- From: "Richard E. Edelmann" {edelmare-at-muohio.edu} 8, 22 -- To: microscopy-at-Microscopy.com 8, 22 -- Date: Wed, 25 Apr 2007 16:42:20 -0400 8, 22 -- MIME-Version: 1.0 8, 22 -- Subject: Question of Terminology - "Microprobe" 8, 22 -- Message-ID: {462F84EC.30245.2A92B38-at-edelmare.muohio.edu} 8, 22 -- Priority: normal 8, 22 -- X-mailer: Pegasus Mail for Windows (4.41) 8, 22 -- Content-type: text/plain; charset=US-ASCII 8, 22 -- Content-transfer-encoding: 7BIT 8, 22 -- Content-description: Mail message body 8, 22 -- X-Scanned-By: MIMEDefang 2.57 on 134.53.6.66 ==============================End of - Headers==============================
I still associate microprobe with the scanning electron beam device equipped with wavelength spectrometers for the express purpose of x-ray microanalysis.
I refer to the little thing used to hold and move objects around as micro-manipulators.
I will admit that micro-probe can refer to a lot of things. I wouldn't be surprised if the microelectronics folks adopted the term to refer to a microscopic test lead. I also suppose the scanning probe microscope folks could refer to their tips as microprobes. But to me, a microprobe will be those things like ARL used to make.
Maybe that makes me old school, but it seems like a decent working definition. I am getting old enough people probably expect me to be a bit pig-headed and crotchety anyway. I might as well live up to it. {g}
Warren Straszheim
-----Original Message----- X-from: edelmare-at-muohio.edu [mailto:edelmare-at-muohio.edu] Sent: Wednesday, April 25, 2007 3:44 PM To: wesaia-at-iastate.edu
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Email: tom-at-farrarlaw.net Name: R. Thomas Farrar
Title-Subject: [Filtered] Water Beading
Question: Within the past year or so I read that scientists had discovered the reason why water beads on lily pads. Recently I have had need for the article (more accurately, for the reason why the water beading occurs) and have searched for it without success. Google led me to this forum, and I wonder whether anyone can help this non-scientist to the people who made the discovery?
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Email: forbes-at-essential-research.com Name: David Forbes
Organization: Essential Research
Title-Subject: [Filtered] ISI Sputter Coater
Question: I'm trying to resurrect an old ISI Sputer Coater Model POL4172 - without a manual. All the components are present, but the operation seems to be questionable. Particularly scary is that when the unit is plugged in, the high voltage is on and plasma strikes right away. It seems that this should wait until I dial on the HV. Any suggestions on troubleshooting or getting a manual would be helpful. I suspect this type of unit is primarily for Au sputtering, but some ideas have been floating around about sputtering more exotic materials (oxides). Any place I can find guidance about sputtering conditions for different materials?
I'd argue that "microprobe" isn't a complete term for anything.
The word "microprobe" can describe dozens of different instruments or devices.
Off the top of my head:
- Electron Microprobe (as in the JEOL type) - Laser Ablation "Microprobe" ICP-MS - Ion "Microprobe" Mass Spectrometer (SIMS) - NASA's new small space probes are called microprobes - Anything with a small sensor or feedback device or manipulator
Consequently, if one is referring to the SEM-like device with WDS, one must call it an electron microprobe, not just a microprobe (except in informal settings where the context is known).
If one is talking about SIMS, one must talk about an ion microprobe. If one is discussing the Deep Space 2 mission, one should call it a space microprobe or something like that. I'd like Warren's term of "micro-manipulators," but one could call them manipulator microprobes or something similar.
Calling the manipulating devices in question "nanoprobes" is not better because that, too, could describe a lot of different devices -- there are already small optical devices called nanoprobes, and the field of "nanoprobes" will only get more crowded in coming years.
Also some, no doubt, will assert that "electron microprobe" isn't the formal term anyway and that the instrument is more properly called an "electron probe microanalyzer" instead. However, if S.J.B. Reed calls it an electron microprobe, then so will I. If anything, raising this point strengthens the above argument. One cannot just call something a "microanalyzer" because that could be dozens or hundreds of devices too. Instead, the full term is needed to describe any instrument properly.
To summarize: "microprobe" really isn't a complete term that describes anything specifically.
Cheers, Ellery
-------------------- Ellery E. Frahm Research Fellow & Manager Electron Microprobe Laboratory University of Minnesota - Twin Cities Department of Geology & Geophysics Alternate e-mail: elleryfrahm-at-mac.com Lab Website: http://probelab.geo.umn.edu Personal Website: http://umn.edu/~frah0010
On Apr 25, 2007, at 3:50 PM, edelmare-at-muohio.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 } ---------------------------------------------------------------------- } ------ } } O.k., folks Lets see if the Microscopy and Microanalysis community } can come an agreement. } } Traditionally the term "Microprobe" has been used to refer to a } dedicated wave length dispersive x-ray microanalysis } microscope/instrument (XWDS or WDS). Is this still a valid use for } the the term "Microprobe"? Or is it antiquated? } } . . . because it comes into conflict a "newer" use of the word } "microprobe" as refering to a "small" physical probing device, } typically mechanical, for manipulating or testing small items. Would } these probing-manipuating-testing devices be better called } "Nanoprobes" (even thought they may actually handle items / objects } in the micrometer to 100's-micrometer range). } } Context will NOT sufice to distinguish between the two as } microscopists now deal with both. } } Anyone care to share comments or opinions ? } } } Richard E. Edelmann, Ph.D. } EXPO Editor, Microscopy and Microanalysis Supplement } Electron Microscopy Facility Director } 364 Pearson Hall } Miami University, Oxford, OH 45056 } Ph: 513.529.5712 Fax: 513.529.4243 } E-mail: edelmare-at-muohio.edu } http://www.emf.muohio.edu
I'm releived that it's not just me who finds 'Electron Probe Micro Analyser' too much of a mouthfull.
But we all use the acronym EPMA, rather than EMP, don't we?
cheers
rtch
On 25 Apr 2007 at 18:31, frah0010-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 } ---------------------------------------------------------------------- } ------ } } Hi all, } } I'd argue that "microprobe" isn't a complete term for anything. } } The word "microprobe" can describe dozens of different instruments or } devices. } } Off the top of my head: } } - Electron Microprobe (as in the JEOL type) } - Laser Ablation "Microprobe" ICP-MS } - Ion "Microprobe" Mass Spectrometer (SIMS) } - NASA's new small space probes are called microprobes } - Anything with a small sensor or feedback device or manipulator } } Consequently, if one is referring to the SEM-like device with WDS, } one must call it an electron microprobe, not just a microprobe } (except in informal settings where the context is known). } } If one is talking about SIMS, one must talk about an ion microprobe. } If one is discussing the Deep Space 2 mission, one should call it a } space microprobe or something like that. I'd like Warren's term of } "micro-manipulators," but one could call them manipulator microprobes } or something similar. } } Calling the manipulating devices in question "nanoprobes" is not } better because that, too, could describe a lot of different devices } -- there are already small optical devices called nanoprobes, and the } field of "nanoprobes" will only get more crowded in coming years. } } Also some, no doubt, will assert that "electron microprobe" isn't the } formal term anyway and that the instrument is more properly called an } "electron probe microanalyzer" instead. However, if S.J.B. Reed } calls it an electron microprobe, then so will I. If anything, } raising this point strengthens the above argument. One cannot just } call something a "microanalyzer" because that could be dozens or } hundreds of devices too. Instead, the full term is needed to } describe any instrument properly. } } To summarize: "microprobe" really isn't a complete term that } describes anything specifically. } } Cheers, } Ellery } } -------------------- } Ellery E. Frahm } Research Fellow & Manager } Electron Microprobe Laboratory } University of Minnesota - Twin Cities } Department of Geology & Geophysics } Alternate e-mail: elleryfrahm-at-mac.com } Lab Website: http://probelab.geo.umn.edu } Personal Website: http://umn.edu/~frah0010 } } } } On Apr 25, 2007, at 3:50 PM, edelmare-at-muohio.edu wrote: } } } -------------------------------------------------------------------- } } -- ------ The Microscopy ListServer -- CoSponsor: The Microscopy } } Society of America To Subscribe/Unsubscribe -- } } http://www.microscopy.com/ MicroscopyListserver On-Line Help
-- 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============================== 10, 27 -- From r.sims-at-auckland.ac.nz Wed Apr 25 18:38:54 2007 10, 27 -- Received: from mailhost.auckland.ac.nz (curly.its.auckland.ac.nz [130.216.10.123]) 10, 27 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3PNcsWi024173 10, 27 -- for {microscopy-at-microscopy.com} ; Wed, 25 Apr 2007 18:38:54 -0500 10, 27 -- Received: from localhost (localhost.localdomain [127.0.0.1]) 10, 27 -- by mailhost.auckland.ac.nz (Postfix) with ESMTP id 270819C2C1 10, 27 -- for {microscopy-at-microscopy.com} ; Thu, 26 Apr 2007 11:38:53 +1200 (NZST) 10, 27 -- X-Virus-Scanned: by amavisd-new at mailhost.auckland.ac.nz 10, 27 -- Received: from mailhost.auckland.ac.nz ([127.0.0.1]) 10, 27 -- by localhost (curly.its.auckland.ac.nz [127.0.0.1]) (amavisd-new, port 10024) 10, 27 -- with ESMTP id BlcN+Qb9F9b6 for {microscopy-at-microscopy.com} ; 10, 27 -- Thu, 26 Apr 2007 11:38:53 +1200 (NZST) 10, 27 -- Received: from rs (r.sims.glg.auckland.ac.nz [130.216.59.18]) 10, 27 -- by mailhost.auckland.ac.nz (Postfix) with ESMTP id F2AA49C29C 10, 27 -- for {microscopy-at-microscopy.com} ; Thu, 26 Apr 2007 11:38:52 +1200 (NZST) 10, 27 -- From: "Ritchie Sims" {r.sims-at-auckland.ac.nz} 10, 27 -- To: microscopy-at-microscopy.com 10, 27 -- Date: Thu, 26 Apr 2007 11:39:43 +1200 10, 27 -- MIME-Version: 1.0 10, 27 -- Subject: Re: [Microscopy] Re: Question of Terminology - "Microprobe" 10, 27 -- Message-ID: {46308F7F.19192.CD2555-at-localhost} 10, 27 -- Priority: normal 10, 27 -- In-reply-to: {200704252331.l3PNVQN7003889-at-ns.microscopy.com} 10, 27 -- X-mailer: Pegasus Mail for Windows (4.21c) 10, 27 -- Content-type: text/plain; charset=US-ASCII 10, 27 -- Content-transfer-encoding: 7BIT 10, 27 -- Content-description: Mail message body ==============================End of - Headers==============================
Hmmmmmmm-If my memory is correct, the Microprobe was invented by Castaing around 1949. I don't know how he spelled it in French, but that was what it was called, and there was little doubt that his Ph.D. thesis describing the instrument and the theoretical physics behind it was nothing less than extraordinary, so please give it the respect it will always be due and don't try to rename it.
John Mardinly
Intel
This comment is not the opinion of Intel Corporation.
-----Original Message----- X-from: edelmare-at-muohio.edu [mailto:edelmare-at-muohio.edu] Sent: Wednesday, April 25, 2007 1:43 PM To: Mardinly, John
O.k., folks Lets see if the Microscopy and Microanalysis community can come an agreement.
Traditionally the term "Microprobe" has been used to refer to a dedicated wave length dispersive x-ray microanalysis microscope/instrument (XWDS or WDS). Is this still a valid use for the the term "Microprobe"? Or is it antiquated?
. . . because it comes into conflict a "newer" use of the word "microprobe" as refering to a "small" physical probing device, typically mechanical, for manipulating or testing small items. Would these probing-manipuating-testing devices be better called "Nanoprobes" (even thought they may actually handle items / objects in the micrometer to 100's-micrometer range).
Context will NOT sufice to distinguish between the two as microscopists now deal with both.
Anyone care to share comments or opinions ?
Richard E. Edelmann, Ph.D. EXPO Editor, Microscopy and Microanalysis Supplement Electron Microscopy Facility Director 364 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu http://www.emf.muohio.edu
==============================Original Headers============================== 8, 22 -- From edelmare-at-muohio.edu Wed Apr 25 15:42:26 2007 8, 22 -- Received: from mulnx11.mcs.muohio.edu (mulnx11.mcs.muohio.edu [134.53.6.66]) 8, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3PKgPg0028602 8, 22 -- for {microscopy-at-Microscopy.com} ; Wed, 25 Apr 2007 15:42:26 -0500 8, 22 -- Received: from mulnx24.mcs.muohio.edu (mulnx24.mcs.muohio.edu [134.53.6.11]) 8, 22 -- by mulnx11.mcs.muohio.edu (Switch-3.1.8/Switch-3.1.7) with ESMTP id l3PKgPSu007140 8, 22 -- for {microscopy-at-Microscopy.com} ; Wed, 25 Apr 2007 16:42:25 -0400 8, 22 -- Received: from [192.168.1.23] ([134.53.14.105]) 8, 22 -- by mulnx24.mcs.muohio.edu (Switch-3.1.8/Switch-3.1.7) with ESMTP id l3PKgOST020652 8, 22 -- for {microscopy-at-Microscopy.com} ; Wed, 25 Apr 2007 16:42:24 -0400 8, 22 -- From: "Richard E. Edelmann" {edelmare-at-muohio.edu} 8, 22 -- To: microscopy-at-Microscopy.com 8, 22 -- Date: Wed, 25 Apr 2007 16:42:20 -0400 8, 22 -- MIME-Version: 1.0 8, 22 -- Subject: Question of Terminology - "Microprobe" 8, 22 -- Message-ID: {462F84EC.30245.2A92B38-at-edelmare.muohio.edu} 8, 22 -- Priority: normal 8, 22 -- X-mailer: Pegasus Mail for Windows (4.41) 8, 22 -- Content-type: text/plain; charset=US-ASCII 8, 22 -- Content-transfer-encoding: 7BIT 8, 22 -- Content-description: Mail message body 8, 22 -- X-Scanned-By: MIMEDefang 2.57 on 134.53.6.66 ==============================End of - Headers==============================
==============================Original Headers============================== 18, 34 -- From john.mardinly-at-intel.com Wed Apr 25 20:36:49 2007 18, 34 -- Received: from mga01.intel.com (mga01.intel.com [192.55.52.88]) 18, 34 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3Q1amga005264 18, 34 -- for {Microscopy-at-msa.microscopy.com} ; Wed, 25 Apr 2007 20:36:49 -0500 18, 34 -- Received: from fmsmga001.fm.intel.com ([10.253.24.23]) 18, 34 -- by fmsmga101.fm.intel.com with ESMTP; 25 Apr 2007 18:36:48 -0700 18, 34 -- X-ExtLoop1: 1 18, 34 -- X-IronPort-AV: E=Sophos;i="4.14,452,1170662400"; 18, 34 -- d="scan'208";a="236867288" 18, 34 -- Received: from fmsmsx333.amr.corp.intel.com ([132.233.42.2]) 18, 34 -- by fmsmga001.fm.intel.com with ESMTP; 25 Apr 2007 18:36:46 -0700 18, 34 -- Received: from scsmsx412.amr.corp.intel.com ([10.3.90.31]) by fmsmsx333.amr.corp.intel.com with Microsoft SMTPSVC(6.0.3790.1830); 18, 34 -- Wed, 25 Apr 2007 18:36:48 -0700 18, 34 -- Received: from scsmsx415.amr.corp.intel.com ([10.3.90.34]) by scsmsx412.amr.corp.intel.com with Microsoft SMTPSVC(6.0.3790.1830); 18, 34 -- Wed, 25 Apr 2007 18:36:47 -0700 18, 34 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 18, 34 -- Content-class: urn:content-classes:message 18, 34 -- MIME-Version: 1.0 18, 34 -- Content-Type: text/plain; 18, 34 -- charset="us-ascii" 18, 34 -- Subject: RE: [Microscopy] Question of Terminology - "Microprobe" 18, 34 -- Date: Wed, 25 Apr 2007 18:36:47 -0700 18, 34 -- Message-ID: {F3CB8931ABF8294DB889E977150CAD685A3D69-at-scsmsx415.amr.corp.intel.com} 18, 34 -- In-Reply-To: {200704252042.l3PKgbtU028724-at-ns.microscopy.com} 18, 34 -- X-MS-Has-Attach: 18, 34 -- X-MS-TNEF-Correlator: 18, 34 -- Thread-Topic: [Microscopy] Question of Terminology - "Microprobe" 18, 34 -- thread-index: AceHekPhB7NnisO8RJmff5J/b5vrnAAJvqKQ 18, 34 -- From: "Mardinly, John" {john.mardinly-at-intel.com} 18, 34 -- To: {edelmare-at-muohio.edu} 18, 34 -- Cc: {Microscopy-at-msa.microscopy.com} 18, 34 -- X-OriginalArrivalTime: 26 Apr 2007 01:36:47.0994 (UTC) FILETIME=[5B06CDA0:01C787A3] 18, 34 -- Content-Transfer-Encoding: 8bit 18, 34 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l3Q1amga005264 ==============================End of - Headers==============================
I have recently received fat pad aspiration biopsies where the fat that was sent is very fine (no more than 20 cells together at it's largest, most are smaller than that.)
If possible, I need a quick method whereby I can take this floating material and condense it to a pellet for embedding in agarose. I do not have the option of hand processing for TEM so transfering this material into a intermediate media is a must.
Thanks in advance,
Mike Ganger
==============================Original Headers============================== 6, 20 -- From mganger-at-optonline.net Wed Apr 25 20:49:16 2007 6, 20 -- Received: from mta4.srv.hcvlny.cv.net (mta4.srv.hcvlny.cv.net [167.206.4.199]) 6, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3Q1nGeV016819 6, 20 -- for {microscopy-at-microscopy.com} ; Wed, 25 Apr 2007 20:49:16 -0500 6, 20 -- Received: from [127.0.0.1] (ool-435145d4.dyn.optonline.net [67.81.69.212]) 6, 20 -- by mta4.srv.hcvlny.cv.net 6, 20 -- (Sun Java System Messaging Server 6.2-6.01 (built Apr 3 2006)) 6, 20 -- with ESMTP id {0JH30044R122UHL0-at-mta4.srv.hcvlny.cv.net} for 6, 20 -- microscopy-at-microscopy.com; Wed, 25 Apr 2007 21:49:16 -0400 (EDT) 6, 20 -- Date: Wed, 25 Apr 2007 21:49:13 -0400 6, 20 -- From: Michael Ganger {mganger-at-optonline.net} 6, 20 -- Subject: TEM help with fat biopsy 6, 20 -- To: microscopy-at-microscopy.com 6, 20 -- Message-id: {46300519.4060402-at-optonline.net} 6, 20 -- MIME-version: 1.0 6, 20 -- Content-type: text/plain; charset=ISO-8859-1; format=flowed 6, 20 -- Content-transfer-encoding: 7BIT 6, 20 -- X-Accept-Language: en-us, en 6, 20 -- User-Agent: Mozilla/5.0 (Windows; U; Windows NT 5.1; en-US; rv:1.7.2) 6, 20 -- Gecko/20040804 Netscape/7.2 (ax) ==============================End of - Headers==============================
My thanks to those that tried to help me find this obscure item from the Norelco Reporter from 40 years ago. I was notified that the article was located at a library and sent for.
This item was more like a newsletter or pamphlet from a company. That made it hard to find, IMO. It appears that it did not end up with Philips Electron Optics (FEI).
Libby, Phil, John, Tom, Ron, and Sharon; my special thanks to all of you.
Paul
==============================Original Headers============================== 6, 26 -- From beaurega-at-westol.com Wed Apr 25 22:19:34 2007 6, 26 -- Received: from smtp-gateway-4.winbeam.com (smtp-gateway-4.winbeam.com [64.84.97.69]) 6, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3Q3JNOo029778 6, 26 -- for {microscopy-at-microscopy.com} ; Wed, 25 Apr 2007 22:19:33 -0500 6, 26 -- Received: from mail.winbeam.com (mail.winbeam.com [64.84.96.10]) 6, 26 -- by smtp-gateway-4.winbeam.com (8.13.2/8.12.8) with SMTP id l3Q3J3WQ005973 6, 26 -- for {microscopy-at-microscopy.com} ; Wed, 25 Apr 2007 23:19:04 -0400 6, 26 -- Received: (qmail 27006 invoked by uid 89); 26 Apr 2007 03:18:45 -0000 6, 26 -- Received: from pitts-69-72-22-249.dynamic-dialup.coretel.net (HELO beaurega) (69.72.22.249) 6, 26 -- by mail.winbeam.com with SMTP; 26 Apr 2007 03:18:45 -0000 6, 26 -- Message-Id: {3.0.6.32.20070425231952.007cc970-at-pop3.norton.antivirus} 6, 26 -- X-Sender: beaurega/mail.westol.com-at-pop3.norton.antivirus 6, 26 -- X-Mailer: QUALCOMM Windows Eudora Light Version 3.0.6 (32) 6, 26 -- Date: Wed, 25 Apr 2007 23:19:52 -0400 6, 26 -- To: microscopy-at-microscopy.com 6, 26 -- From: Beaurega {beaurega-at-westol.com} 6, 26 -- Subject: Re: [Microscopy] Request for 1964 Norelco Reporter Article or 6, 26 -- Page 6, 26 -- In-Reply-To: {p06230901c25486630d07-at-[192.168.0.3]} 6, 26 -- Mime-Version: 1.0 6, 26 -- Content-Type: text/plain; charset="us-ascii" 6, 26 -- X-smtp-gateway-4-MailScanner-Information: smtp-gateway-4 - Please contact Technical Support for more information 6, 26 -- X-smtp-gateway-4-MailScanner: Found to be clean smtp-gateway-4 (courtesy of MailScanner) 6, 26 -- X-smtp-gateway-4-MailScanner-SpamCheck: not spam (whitelisted), 6, 26 -- SpamAssassin (not cached, score=0.992, required 4, AWL 0.99) 6, 26 -- X-smtp-gateway-4-MailScanner-From: beaurega-at-westol.com ==============================End of - Headers==============================
On Apr 25, 2007, at 8:42 PM, john.mardinly-at-intel.com wrote: } } Hmmmmmmm-If my memory is correct, the Microprobe was invented by } Castaing around 1949. I don't know how he spelled it in French, but } that } was what it was called, and there was little doubt that his Ph.D. } thesis } describing the instrument and the theoretical physics behind it was } nothing less than extraordinary, so please give it the respect it will } always be due and don't try to rename it. } } John Mardinly
Actually Castaing's 1951 thesis was titled "Application des Sondes Electroniques a Une Methode d'Analyse Poncteculle Chimique et Cristallographique" which roughly translates as "Application of Electron Probes as a Method of Local Chemical and Crystallographic Analysis." I'm not sure what he called his device in the thesis -- I don't have a copy in French to check. Anyone?
I know that Castaing's 1953 paper with Guinier in "Analytical Chemistry" calls it the "electronic microanalyzer." I believe, though, when Cameca was developing their first commercial instrument, they were using the word "microsonde" to describe it.
I'm not certain what Borovskii called his device in 1953. Nor do I know what Hillier called the device he patented in 1947 -- the title of his patent was something like "electron probe analysis employing x- ray spectrography," I believe.
I don't think we should feel too obligated to use Castaing's terminology (despite the problems presented by "microsonde" or "electronic analyzer"). In science, we should be able to reclassify things based on new definitions or better categories, such as redefining planets to include extrasolar bodies but exclude Pluto and other Kuiper Belt objects.
I do agree with John, though, because I don't want to order new stationary.
Best, Ellery
-------------------- Ellery E. Frahm Research Fellow & Manager Electron Microprobe Laboratory University of Minnesota - Twin Cities Department of Geology & Geophysics Alternate e-mail: elleryfrahm-at-mac.com Lab Website: http://probelab.geo.umn.edu Personal Website: http://umn.edu/~frah0010
} I'm releived that it's not just me who finds 'Electron Probe } Micro Analyser' too much of a mouthfull. } } But we all use the acronym EPMA, rather than EMP, don't we?
If that's a question and this is a poll, I'm in agreement. "EPMA" is the defacto acronym, but there's no end to the number of us who still refer to it also as a "microprobe".
Genuinely, Michael Shaffer :o)
SEM/MLA Research Coordinator http://www.mun.ca/creait/maf/
Inco Innovation Centre Memorial University St. John's, Newfoundland
==============================Original Headers============================== 8, 20 -- From michael-at-shaffer.net Thu Apr 26 04:46:30 2007 8, 20 -- Received: from n007.sc1.he.tucows.com (smtpout1458.sc1.he.tucows.com [64.97.157.158]) 8, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3Q9kTUg027760 8, 20 -- for {Microscopy-at-microscopy.com} ; Thu, 26 Apr 2007 04:46:30 -0500 8, 20 -- Received: from rarewolf (205.251.83.78) by n007.sc1.he.tucows.com (7.2.069.1) (authenticated as michael-at-shaffer.net) 8, 20 -- id 4620B52300213E46 for Microscopy-at-microscopy.com; Thu, 26 Apr 2007 09:46:27 +0000 8, 20 -- From: "michael shaffer" {michael-at-shaffer.net} 8, 20 -- To: "MSA Microscopy list" {Microscopy-at-microscopy.com} 8, 20 -- References: {200704252339.l3PNdEsu024860-at-ns.microscopy.com} 8, 20 -- Subject: RE: [Microscopy] Question of Terminology - "Microprobe" 8, 20 -- Date: Thu, 26 Apr 2007 07:16:19 -0230 8, 20 -- Message-ID: {00d101c787e7$beaca890$4701a8c0-at-rarewolf} 8, 20 -- MIME-Version: 1.0 8, 20 -- Content-Type: text/plain; 8, 20 -- charset="us-ascii" 8, 20 -- Content-Transfer-Encoding: 7bit 8, 20 -- X-Mailer: Microsoft Office Outlook 11 8, 20 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.3028 8, 20 -- Thread-Index: AceHknWFqn77gLJKTsa5rc9Bv50f5wAVK9Iw 8, 20 -- In-Reply-To: {200704252339.l3PNdEsu024860-at-ns.microscopy.com} ==============================End of - Headers==============================
From lucidic-at-virtual2.webair.com Thu Apr 26 05:28:32 2007 Return-Path: {lucidic-at-virtual2.webair.com} Received: from virtual2.webair.com (tony.webair.com [216.130.161.119]) by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3QASVEu007712 for {MicroscopyListserverArchive-at-microscopy.com} ; Thu, 26 Apr 2007 05:28:31 -0500 Received: from virtual2.webair.com (localhost.webair.com [127.0.0.1]) by virtual2.webair.com (8.13.8/8.12.11) with ESMTP id l3QASW0g086504 for {MicroscopyListserverArchive-at-microscopy.com} ; Thu, 26 Apr 2007 06:28:33 -0400 (EDT) (envelope-from lucidic-at-virtual2.webair.com) Received: (from lucidic-at-localhost) by virtual2.webair.com (8.13.8/8.12.11/Submit) id l3QASWaR086420; Thu, 26 Apr 2007 06:28:32 -0400 (EDT) (envelope-from lucidic)
I have to agree with Warren, especially perhaps his last paragraph. "Microprobe" has a definite meaning to me and most microscopists. I wouldn't suggest using "nanoprobe", as that doesn't really solve the problem of confusing terms. Perhaps a simple addition of an adjective to make it a noun phrase, like "mechanical microprobe". Micromanipulator is a well-known term, so the next step would be nanomanipulator. Let's start using that before some company trademarks it so no one else can use it.
Phil
} I still associate microprobe with the scanning electron beam device } equipped with wavelength spectrometers for the express purpose of x-ray } microanalysis. } } I refer to the little thing used to hold and move objects around as } micro-manipulators. } } I will admit that micro-probe can refer to a lot of things. I wouldn't } be surprised if the microelectronics folks adopted the term to refer to } a microscopic test lead. I also suppose the scanning probe microscope } folks could refer to their tips as microprobes. But to me, a microprobe } will be those things like ARL used to make. } } Maybe that makes me old school, but it seems like a decent working } definition. I am getting old enough people probably expect me to be a } bit pig-headed and crotchety anyway. I might as well live up to it. {g} } } Warren Straszheim } } } -----Original Message----- } X-from: edelmare-at-muohio.edu [mailto:edelmare-at-muohio.edu] } Sent: Wednesday, April 25, 2007 3:44 PM } To: wesaia-at-iastate.edu } Subject: [Microscopy] Question of Terminology - "Microprobe" } } } O.k., folks Lets see if the Microscopy and Microanalysis community } can come an agreement. } } Traditionally the term "Microprobe" has been used to refer to a } dedicated wave length dispersive x-ray microanalysis } microscope/instrument (XWDS or WDS). Is this still a valid use for } the the term "Microprobe"? Or is it antiquated? } } . . . because it comes into conflict a "newer" use of the word } "microprobe" as refering to a "small" physical probing device, } typically mechanical, for manipulating or testing small items. Would } these probing-manipuating-testing devices be better called } "Nanoprobes" (even thought they may actually handle items / objects } in the micrometer to 100's-micrometer range). } } Context will NOT sufice to distinguish between the two as } microscopists now deal with both. } } Anyone care to share comments or opinions ? } } } Richard E. Edelmann, Ph.D. } EXPO Editor, Microscopy and Microanalysis Supplement } Electron Microscopy Facility Director } 364 Pearson Hall } Miami University, Oxford, OH 45056 } Ph: 513.529.5712 Fax: 513.529.4243 } E-mail: edelmare-at-muohio.edu } http://www.emf.muohio.edu -- 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============================== 3, 22 -- From oshel1pe-at-cmich.edu Thu Apr 26 07:56:59 2007 3, 22 -- Received: from ob2.cmich.edu (ob2.cmich.edu [141.209.20.21]) 3, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3QCuxCm010367 3, 22 -- for {Microscopy-at-microscopy.com} ; Thu, 26 Apr 2007 07:56:59 -0500 3, 22 -- Received: from egatea.central.cmich.local ([141.209.15.74]) 3, 22 -- by ob2.cmich.edu (8.12.10/8.12.10) with ESMTP id l3QDL26h026847 3, 22 -- for {Microscopy-at-microscopy.com} ; Thu, 26 Apr 2007 09:21:02 -0400 3, 22 -- Received: from [141.209.160.249] ([141.209.160.249]) by egatea.central.cmich.local with Microsoft SMTPSVC(6.0.3790.1830); 3, 22 -- Thu, 26 Apr 2007 08:56:56 -0400 3, 22 -- Mime-Version: 1.0 3, 22 -- Message-Id: {f06230904c25650f2b54c-at-[141.209.160.249]} 3, 22 -- In-Reply-To: {200704252113.l3PLDaRo014950-at-ns.microscopy.com} 3, 22 -- References: {200704252113.l3PLDaRo014950-at-ns.microscopy.com} 3, 22 -- Date: Thu, 26 Apr 2007 08:56:55 -0400 3, 22 -- To: Microscopy-at-microscopy.com 3, 22 -- From: Philip Oshel {oshel1pe-at-cmich.edu} 3, 22 -- Subject: [Microscopy] RE: Question of Terminology - "Microprobe" 3, 22 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 3, 22 -- X-OriginalArrivalTime: 26 Apr 2007 12:56:56.0194 (UTC) FILETIME=[5E9BF620:01C78802] 3, 22 -- X-CanItPRO-Stream: default 3, 22 -- X-Spam-Score: -3.7 () L_EXCH_MF,MY_HTML_OBFU 3, 22 -- X-Scanned-By: CanIt (www . roaringpenguin . com) on 141.209.20.21 ==============================End of - Headers==============================
Ellery pretty much captured what my response would be. "Microprobe" by itself is too generic. The electron microprobe has been, and still is, called many things. Electron probe, microprobe, electron microprobe, "the" probe, etc. I have colleagues who even call it "the WDS", but that's a different problem. I think the "electron microprobe" is the most commonly used and accepted term for the instrument, at least in the countless number of reports that included technique descriptions in the geological literature.
EPMA is a widely accepted acronym, used both for the technique (microanalysis) and instrument (microanalyzer). I prefer to use it for the technique and describe the instrument as the electron microprobe.
Jim McGee ************************************ James J. McGee Materials Engineer Lockheed Martin ************************************ ----- Original Message ----- X-from: {frah0010-at-umn.edu} To: {mcgeejj-at-kapl.gov} Sent: Wednesday, April 25, 2007 7:36 PM
All,
Our Gatan 622 SIT camera died. The 4000 is too old to invest in a new camera or even repair the dead one. I've heard that many 4000's have been decommissioned, do any of you have the 622 camera or the wide-angle CCD camera (model 673) you'd sell, trade, donate?
Owen Mills Michigan Tech Univ.
==============================Original Headers============================== 3, 31 -- From opmills-at-mtu.edu Thu Apr 26 09:36:21 2007 3, 31 -- Received: from node8.mtu.edu (node8.mtu.edu [141.219.69.8]) 3, 31 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3QEaLmh003285 3, 31 -- for {Microscopy-at-microscopy.com} ; Thu, 26 Apr 2007 09:36:21 -0500 3, 31 -- Received: from node6.edge.dcsint.mtu.edu (node6.mtu.edu [141.219.69.6]) 3, 31 -- by node8.mtu.edu (8.13.1/8.13.1) with ESMTP id l3QEaLiv010757 3, 31 -- for {Microscopy-at-microscopy.com} ; Thu, 26 Apr 2007 10:36:21 -0400 3, 31 -- Received: from node34.edge.dcsint.mtu.edu (node34.mtu.edu [141.219.69.34]) 3, 31 -- by node6.edge.dcsint.mtu.edu (8.12.11.20060308/8.12.8) with ESMTP id l3QEaLDZ002228 3, 31 -- for {Microscopy-at-microscopy.com} ; Thu, 26 Apr 2007 10:36:21 -0400 3, 31 -- Received: from mail.mtu.edu (campus0.mtu.edu [141.219.70.8]) 3, 31 -- by node34.edge.dcsint.mtu.edu (8.13.8/8.13.8) with ESMTP id l3QEaKnh017349 3, 31 -- for {Microscopy-at-microscopy.com} ; Thu, 26 Apr 2007 10:36:20 -0400 3, 31 -- (envelope-from opmills-at-mtu.edu) 3, 31 -- Received: from node5.edge.dcsint.mtu.edu (node5.mtu.edu [141.219.69.5]) 3, 31 -- by mail.mtu.edu (8.13.6+Sun/8.11.6) with ESMTP id l3QEaKis016266 3, 31 -- for {Microscopy-at-microscopy.com} ; Thu, 26 Apr 2007 10:36:20 -0400 (EDT) 3, 31 -- Received: from [141.219.192.45] (opmills.eecn.sabo.mtu.edu [141.219.192.45]) 3, 31 -- by node5.edge.dcsint.mtu.edu (8.12.11.20060308/8.12.3/auth_ssl.mc v1.0) with ESMTP id l3QEaKKB031762 3, 31 -- for {Microscopy-at-microscopy.com} ; Thu, 26 Apr 2007 10:36:20 -0400 3, 31 -- Mime-Version: 1.0 (Apple Message framework v752.3) 3, 31 -- Content-Transfer-Encoding: 7bit 3, 31 -- Message-Id: {0B28A29B-B351-4A63-BA41-9E87A04DA8BD-at-mtu.edu} 3, 31 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 3, 31 -- To: Microscopy Listserver {Microscopy-at-microscopy.com} 3, 31 -- From: "Owen P. Mills" {opmills-at-mtu.edu} 3, 31 -- Subject: wanted old Jeol 4000 TEM camera 3, 31 -- Date: Thu, 26 Apr 2007 10:36:18 -0400 3, 31 -- X-Mailer: Apple Mail (2.752.3) 3, 31 -- X-PMX-Version: 5.3.1.294258, Antispam-Engine: 2.5.0.283055, Antispam-Data: 2007.4.7.54434 3, 31 -- X-PerlMx-Spam: Gauge=IIIIIII, Probability=7%, Report='__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' ==============================End of - Headers==============================
Our lab needs to purchase a new scanner for our TEM negatives. We have narrowed our choices to either the Epson Perfection V750 (recommended by Keith Young in the May 2006 issue of Microscopy Today) or the Microtek ScanMaker i900. The major difference between the two seems to be the "glassless" scanning feature of the Microtek. Has anyone had first-hand experience of the film scanning ability of either one of these scanners? Or are there any quirks that would make one of them less desirable?
Thank you, Julie Gross Kent Morest Lab Dept. of Neuroscience UCONN Health Center Farmington, CT 06030 jgross-at-neuron.uchc.edu
==============================Original Headers============================== 3, 18 -- From jgross-at-neuron.uchc.edu Thu Apr 26 13:50:49 2007 3, 18 -- Received: from uchc.edu (itiron1.uchc.edu [155.37.201.25]) 3, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3QIomws020028 3, 18 -- for {microscopy-at-microscopy.com} ; Thu, 26 Apr 2007 13:50:49 -0500 3, 18 -- Received: from ([155.37.91.71]) 3, 18 -- by itiron1.uchc.edu with ESMTP id 5502373.19449977; 3, 18 -- Thu, 26 Apr 2007 14:50:39 -0400 3, 18 -- Received: from morgpc.neuron.uchc.edu ([155.37.1.96]) by itnwa.uchc.edu with Microsoft SMTPSVC(5.0.2195.6713); 3, 18 -- Thu, 26 Apr 2007 14:50:38 -0400 3, 18 -- X-Mailer: QUALCOMM Windows Eudora Version 7.1.0.9 3, 18 -- Date: Thu, 26 Apr 2007 14:50:38 -0400 3, 18 -- To: microscopy-at-microscopy.com 3, 18 -- From: Julie Gross {jgross-at-neuron.uchc.edu} 3, 18 -- Subject: Scanners for TEM negatives 3, 18 -- Mime-Version: 1.0 3, 18 -- Content-Type: text/plain; charset="us-ascii"; format=flowed 3, 18 -- Message-ID: {ITNWA6PYzDabigPasSm00000115-at-itnwa.uchc.edu} 3, 18 -- X-OriginalArrivalTime: 26 Apr 2007 18:50:39.0083 (UTC) FILETIME=[C86FD3B0:01C78833] ==============================End of - Headers==============================
I solved this problem by putting my negatives, emulsion side up, on a light box, masking off stray light, and photographing with my digital camera (3 year old Canon G3) at maximum resolution. I 'invert' the image with PhotoShop to get a positive and adjust contrast as needed. Works great and a lot faster than a scanner.
Geoff
jgross-at-neuron.uchc.edu wrote:
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==============================Original Headers============================== 6, 34 -- From mcauliff-at-umdnj.edu Thu Apr 26 14:09:39 2007 6, 34 -- Received: from zix01.umdnj.edu (zix01.UMDNJ.EDU [130.219.34.124]) 6, 34 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3QJ9cbp032018 6, 34 -- for {microscopy-at-msa.microscopy.com} ; Thu, 26 Apr 2007 14:09:39 -0500 6, 34 -- Received: from zix01.umdnj.edu (ZixVPM [127.0.0.1]) 6, 34 -- by Outbound.umdnj.edu (Proprietary) with ESMTP id C8BBBA7C2E 6, 34 -- for {microscopy-at-msa.microscopy.com} ; Thu, 26 Apr 2007 15:09:37 -0400 (EDT) 6, 34 -- Received: from umdnj.edu (imail2.UMDNJ.EDU [130.219.34.139]) 6, 34 -- by zix01.umdnj.edu (Proprietary) with ESMTP id C1B09A7B82 6, 34 -- for {microscopy-at-msa.microscopy.com} ; Thu, 26 Apr 2007 15:09:36 -0400 (EDT) 6, 34 -- Received: from ([130.219.34.131]) 6, 34 -- by imail2.umdnj.edu with ESMTP id KP-BRCRF.97318950; 6, 34 -- Thu, 26 Apr 2007 15:09:17 -0400 6, 34 -- Received: from conversion-daemon.Polaris.umdnj.edu by Polaris.umdnj.edu 6, 34 -- (iPlanet Messaging Server 5.2 HotFix 2.02 (built Oct 21 2004)) 6, 34 -- id {0JH400M01D291Z-at-Polaris.umdnj.edu} (original mail from mcauliff-at-umdnj.edu) 6, 34 -- for microscopy-at-msa.microscopy.com; Thu, 26 Apr 2007 15:09:17 -0400 (EDT) 6, 34 -- Received: from [127.0.0.1] ([10.138.2.123]) 6, 34 -- by Polaris.umdnj.edu (iPlanet Messaging Server 5.2 HotFix 2.02 (built Oct 21 6, 34 -- 2004)) with ESMTP id {0JH400A4FD7GLZ-at-Polaris.umdnj.edu} ; Thu, 6, 34 -- 26 Apr 2007 15:09:17 -0400 (EDT) 6, 34 -- Date: Thu, 26 Apr 2007 15:10:07 -0400 6, 34 -- From: Geoff McAuliffe {mcauliff-at-umdnj.edu} 6, 34 -- Subject: Re: [Microscopy] Scanners for TEM negatives 6, 34 -- In-reply-to: {200704261852.l3QIqBP6021852-at-ns.microscopy.com} 6, 34 -- To: jgross-at-neuron.uchc.edu, microscopy-at-msa.microscopy.com 6, 34 -- Message-id: {4630F90F.7040300-at-umdnj.edu} 6, 34 -- MIME-version: 1.0 6, 34 -- Content-type: text/plain; format=flowed; charset=ISO-8859-1 6, 34 -- Content-transfer-encoding: 7BIT 6, 34 -- X-Accept-Language: en-us, en 6, 34 -- User-Agent: Mozilla/5.0 (Windows; U; Windows NT 5.1; en-US; rv:1.7.2) 6, 34 -- Gecko/20040804 Netscape/7.2 (ax) 6, 34 -- References: {200704261852.l3QIqBP6021852-at-ns.microscopy.com} ==============================End of - Headers==============================
I would recommend the Nikon Coolscan. http://www.nikoncoolscan.com . You can probably find one for very cheap on ebay or craigslist. Shawn
-- Shawn Mikula, Ph.D., Postdoctoral Scholar Center for Neuroscience University of California-Davis, 1544 Newton Court, Davis, CA 95618, Phone: 530-754-9209 Fax: 530-754-9136 mail: samikula-at-ucdavis.edu web: http://brainmaps.org
----- Original Message ----- X-from: {jgross-at-neuron.uchc.edu} To: {samikula-at-ucdavis.edu} Sent: Thursday, April 26, 2007 11:58 AM
Julie, I do not know about these scanners yet I would try them at your local stores before buying either one, look at the outcome.
Ani
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==============================Original Headers============================== 4, 28 -- From ani.issaian-at-csun.edu Thu Apr 26 14:14:29 2007 4, 28 -- Received: from plover.csun.edu (plover.csun.edu [130.166.1.24]) 4, 28 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3QJESFk011258 4, 28 -- for {microscopy-at-microscopy.com} ; Thu, 26 Apr 2007 14:14:29 -0500 4, 28 -- Received: from puffin.csun.edu (puffin.csun.edu [130.166.1.21]) 4, 28 -- by plover.csun.edu (MOS 3.8.2-GA) 4, 28 -- with ESMTP id DSQ74436; 4, 28 -- Thu, 26 Apr 2007 12:14:27 -0700 (PDT) 4, 28 -- Received: from cuckoo.csun.edu (cuckoo.csun.edu [130.166.1.230]) 4, 28 -- by puffin.csun.edu (MOS 3.7.5a-GA) 4, 28 -- with ESMTP id FRH77806; 4, 28 -- Thu, 26 Apr 2007 12:14:25 -0700 (PDT) 4, 28 -- Received: (from cuckoo.csun.edu [130.166.114.86]) 4, 28 -- by cuckoo.csun.edu (MOS 3.8.2-GA) 4, 28 -- with HTTPS/1.1 id BAN69426 (AUTH ami24015); 4, 28 -- Thu, 26 Apr 2007 12:14:25 -0700 (PDT) 4, 28 -- From: Ani M Issaian {ani.issaian-at-csun.edu} 4, 28 -- Subject: Re: [Microscopy] Scanners for TEM negatives 4, 28 -- To: jgross-at-neuron.uchc.edu 4, 28 -- Cc: microscopy-at-microscopy.com 4, 28 -- Reply-To: ani.issaian-at-csun.edu 4, 28 -- X-Mailer: Mirapoint Webmail Direct 3.8.2-GA 4, 28 -- MIME-Version: 1.0 4, 28 -- Content-Type: text/plain; charset=us-ascii 4, 28 -- Content-Transfer-Encoding: 7bit 4, 28 -- Message-Id: {20070426121425.BAN69426-at-cuckoo.csun.edu} 4, 28 -- Date: Thu, 26 Apr 2007 12:14:25 -0700 (PDT) 4, 28 -- X-Junkmail-Whitelist: YES (by domain whitelist at plover.csun.edu) ==============================End of - Headers==============================
O.k. it sounds fairly unamimous that "Electron Microprobe / EPMA" is the genrally used accepted terminology for a "WDS-Microprobe" and is better than simply microprobe.
BUT Phil hits the nail on the head with regards to "mechanical microprobe" type devices.
==} "Let's start using [a term] before some company trademarks it so no one else can use it."
So what shall it be folks? "Mechanical Microprobe"? "Nanomanipulator"? Something else?
Let us the microscopists decide before someone trademarks it.
} } I have to agree with Warren, especially perhaps his last paragraph. } "Microprobe" has a definite meaning to me and most microscopists. } I wouldn't suggest using "nanoprobe", as that doesn't really solve } the problem of confusing terms. Perhaps a simple addition of an } adjective to make it a noun phrase, like "mechanical microprobe". } Micromanipulator is a well-known term, so the next step would be } nanomanipulator. Let's start using that before some company } trademarks it so no one else can use it. } } Phil } } } I still associate microprobe with the scanning electron beam device } } equipped with wavelength spectrometers for the express purpose of x-ray } } microanalysis. } } } } I refer to the little thing used to hold and move objects around as } } micro-manipulators. } } } } I will admit that micro-probe can refer to a lot of things. I wouldn't } } be surprised if the microelectronics folks adopted the term to refer to } } a microscopic test lead. I also suppose the scanning probe microscope } } folks could refer to their tips as microprobes. But to me, a microprobe } } will be those things like ARL used to make. } } } } Maybe that makes me old school, but it seems like a decent working } } definition. I am getting old enough people probably expect me to be a } } bit pig-headed and crotchety anyway. I might as well live up to it. {g} } } } } Warren Straszheim } } } } } } -----Original Message----- } } X-from: edelmare-at-muohio.edu [mailto:edelmare-at-muohio.edu] } } Sent: Wednesday, April 25, 2007 3:44 PM } } To: wesaia-at-iastate.edu } } Subject: [Microscopy] Question of Terminology - "Microprobe" } } } } } } O.k., folks Lets see if the Microscopy and Microanalysis community } } can come an agreement. } } } } Traditionally the term "Microprobe" has been used to refer to a } } dedicated wave length dispersive x-ray microanalysis } } microscope/instrument (XWDS or WDS). Is this still a valid use for } } the the term "Microprobe"? Or is it antiquated? } } } } . . . because it comes into conflict a "newer" use of the word } } "microprobe" as refering to a "small" physical probing device, } } typically mechanical, for manipulating or testing small items. Would } } these probing-manipuating-testing devices be better called } } "Nanoprobes" (even thought they may actually handle items / objects } } in the micrometer to 100's-micrometer range). } } } } Context will NOT sufice to distinguish between the two as } } microscopists now deal with both. } } } } Anyone care to share comments or opinions ? } } } } } } Richard E. Edelmann, Ph.D. } } EXPO Editor, Microscopy and Microanalysis Supplement } } Electron Microscopy Facility Director } } 364 Pearson Hall } } Miami University, Oxford, OH 45056 } } Ph: 513.529.5712 Fax: 513.529.4243 } } E-mail: edelmare-at-muohio.edu } } http://www.emf.muohio.edu } -- } 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============================== } 3, 22 -- From oshel1pe-at-cmich.edu Thu Apr 26 07:56:59 2007 } 3, 22 -- Received: from ob2.cmich.edu (ob2.cmich.edu [141.209.20.21]) } 3, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3QCuxCm010367 } 3, 22 -- for {Microscopy-at-microscopy.com} ; Thu, 26 Apr 2007 07:56:59 -0500 } 3, 22 -- Received: from egatea.central.cmich.local ([141.209.15.74]) } 3, 22 -- by ob2.cmich.edu (8.12.10/8.12.10) with ESMTP id l3QDL26h026847 } 3, 22 -- for {Microscopy-at-microscopy.com} ; Thu, 26 Apr 2007 09:21:02 -0400 } 3, 22 -- Received: from [141.209.160.249] ([141.209.160.249]) by egatea.central.cmich.local with Microsoft SMTPSVC(6.0.3790.1830); } 3, 22 -- Thu, 26 Apr 2007 08:56:56 -0400 } 3, 22 -- Mime-Version: 1.0 } 3, 22 -- Message-Id: {f06230904c25650f2b54c-at-[141.209.160.249]} } 3, 22 -- In-Reply-To: {200704252113.l3PLDaRo014950-at-ns.microscopy.com} } 3, 22 -- References: {200704252113.l3PLDaRo014950-at-ns.microscopy.com} } 3, 22 -- Date: Thu, 26 Apr 2007 08:56:55 -0400 } 3, 22 -- To: Microscopy-at-microscopy.com } 3, 22 -- From: Philip Oshel {oshel1pe-at-cmich.edu} } 3, 22 -- Subject: [Microscopy] RE: Question of Terminology - "Microprobe" } 3, 22 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" } 3, 22 -- X-OriginalArrivalTime: 26 Apr 2007 12:56:56.0194 (UTC) FILETIME=[5E9BF620:01C78802] } 3, 22 -- X-CanItPRO-Stream: default } 3, 22 -- X-Spam-Score: -3.7 () L_EXCH_MF,MY_HTML_OBFU } 3, 22 -- X-Scanned-By: CanIt (www . roaringpenguin . com) on 141.209.20.21 } ==============================End of - Headers==============================
Richard E. Edelmann, Ph.D. EXPO Editor, Microscopy and Microanalysis Supplement Electron Microscopy Facility Director 364 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu http://www.emf.muohio.edu
==============================Original Headers============================== 9, 24 -- From edelmare-at-muohio.edu Thu Apr 26 14:53:04 2007 9, 24 -- Received: from mulnx12.mcs.muohio.edu (mulnx12.mcs.muohio.edu [134.53.6.67]) 9, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3QJr371002286 9, 24 -- for {microscopy-at-Microscopy.com} ; Thu, 26 Apr 2007 14:53:04 -0500 9, 24 -- Received: from mulnx23.mcs.muohio.edu (mulnx23.mcs.muohio.edu [134.53.6.10]) 9, 24 -- by mulnx12.mcs.muohio.edu (Switch-3.1.8/Switch-3.1.7) with ESMTP id l3QJr2IP013948 9, 24 -- for {microscopy-at-Microscopy.com} ; Thu, 26 Apr 2007 15:53:02 -0400 9, 24 -- Received: from [192.168.1.23] ([134.53.14.105]) 9, 24 -- by mulnx23.mcs.muohio.edu (Switch-3.1.8/Switch-3.1.7) with ESMTP id l3QJr25X002076 9, 24 -- for {microscopy-at-Microscopy.com} ; Thu, 26 Apr 2007 15:53:02 -0400 9, 24 -- From: "Richard E. Edelmann" {edelmare-at-muohio.edu} 9, 24 -- To: microscopy-at-Microscopy.com 9, 24 -- Date: Thu, 26 Apr 2007 15:53:02 -0400 9, 24 -- MIME-Version: 1.0 9, 24 -- Subject: Re: Terminology - please vote! 9, 24 -- Message-ID: {4630CADE.22716.27D1525-at-edelmare.muohio.edu} 9, 24 -- Priority: normal 9, 24 -- In-reply-to: {200704261258.l3QCw7lS011622-at-ns.microscopy.com} 9, 24 -- References: {200704261258.l3QCw7lS011622-at-ns.microscopy.com} 9, 24 -- X-mailer: Pegasus Mail for Windows (4.41) 9, 24 -- Content-type: text/plain; charset=US-ASCII 9, 24 -- Content-transfer-encoding: 7BIT 9, 24 -- Content-description: Mail message body 9, 24 -- X-Scanned-By: MIMEDefang 2.57 on 134.53.6.67 ==============================End of - Headers==============================
Julie, I have what is now and old Epson Expression 1600 Professional scanner with trans-illumiator. It works well and is easy. I would imagine that the newer generation from Epson is also good. Lee -- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy & Histology Core Facility Manager, Optical Microscopy Core Facility Weill-Cornell Medical College
Microtek i900. Hands down. No questions asked! Great intuitive interface. Been a Microtek fan for years. Bet regards, Walt
-----Original Message----- X-from: jgross-at-neuron.uchc.edu [mailto:jgross-at-neuron.uchc.edu] Sent: Thursday, April 26, 2007 2:57 PM To: Bobrowski, Walter
Our lab needs to purchase a new scanner for our TEM negatives. We have narrowed our choices to either the Epson Perfection V750 (recommended by Keith Young in the May 2006 issue of Microscopy Today) or the Microtek ScanMaker i900. The major difference between the two seems to be the "glassless" scanning feature of the Microtek. Has anyone had first-hand experience of the film scanning ability of either one of these scanners? Or are there any quirks that would make one of them less desirable?
Thank you, Julie Gross Kent Morest Lab Dept. of Neuroscience UCONN Health Center Farmington, CT 06030 jgross-at-neuron.uchc.edu
==============================Original Headers============================== 3, 18 -- From jgross-at-neuron.uchc.edu Thu Apr 26 13:50:49 2007 3, 18 -- Received: from uchc.edu (itiron1.uchc.edu [155.37.201.25]) 3, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3QIomws020028 3, 18 -- for {microscopy-at-microscopy.com} ; Thu, 26 Apr 2007 13:50:49 -0500 3, 18 -- Received: from ([155.37.91.71]) 3, 18 -- by itiron1.uchc.edu with ESMTP id 5502373.19449977; 3, 18 -- Thu, 26 Apr 2007 14:50:39 -0400 3, 18 -- Received: from morgpc.neuron.uchc.edu ([155.37.1.96]) by itnwa.uchc.edu with Microsoft SMTPSVC(5.0.2195.6713); 3, 18 -- Thu, 26 Apr 2007 14:50:38 -0400 3, 18 -- X-Mailer: QUALCOMM Windows Eudora Version 7.1.0.9 3, 18 -- Date: Thu, 26 Apr 2007 14:50:38 -0400 3, 18 -- To: microscopy-at-microscopy.com 3, 18 -- From: Julie Gross {jgross-at-neuron.uchc.edu} 3, 18 -- Subject: Scanners for TEM negatives 3, 18 -- Mime-Version: 1.0 3, 18 -- Content-Type: text/plain; charset="us-ascii"; format=flowed 3, 18 -- Message-ID: {ITNWA6PYzDabigPasSm00000115-at-itnwa.uchc.edu} 3, 18 -- X-OriginalArrivalTime: 26 Apr 2007 18:50:39.0083 (UTC) FILETIME=[C86FD3B0:01C78833] ==============================End of - Headers==============================
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==============================Original Headers============================== 12, 31 -- From Walter.Bobrowski-at-pfizer.com Thu Apr 26 15:05:51 2007 12, 31 -- Received: from gromsgoa01.pfizer.com (gromsgo.pfizer.com [148.168.224.84]) 12, 31 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3QK5oqQ025306 12, 31 -- for {microscopy-at-microscopy.com} ; Thu, 26 Apr 2007 15:05:51 -0500 12, 31 -- Received: from mopamrexc02.amer.pfizer.com (mopamrexc02.pfizer.com [170.116.30.68]) 12, 31 -- by gromsgoa01.pfizer.com (8.13.7/8.13.7) with ESMTP id l3QK5o7X012707; 12, 31 -- Thu, 26 Apr 2007 16:05:50 -0400 12, 31 -- Received: from mopamrexc01.amer.pfizer.com ([170.116.32.254]) by mopamrexc02.amer.pfizer.com with Microsoft SMTPSVC(6.0.3790.1830); 12, 31 -- Thu, 26 Apr 2007 16:05:50 -0400 12, 31 -- Received: from anaamrexm01.amer.pfizer.com ([162.48.101.10]) by mopamrexc01.amer.pfizer.com with Microsoft SMTPSVC(6.0.3790.1830); 12, 31 -- Thu, 26 Apr 2007 16:05:49 -0400 12, 31 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 12, 31 -- Content-class: urn:content-classes:message 12, 31 -- MIME-Version: 1.0 12, 31 -- Content-Type: text/plain; 12, 31 -- charset="iso-8859-1" 12, 31 -- Subject: RE: [Microscopy] Scanners for TEM negatives 12, 31 -- Date: Thu, 26 Apr 2007 16:05:48 -0400 12, 31 -- Message-ID: {CF89AF6ABD93B746A55BCCD9480A4B3D044EC285-at-anaamrexm01.amer.pfizer.com} 12, 31 -- In-Reply-To: {200704261857.l3QIv0Eq028929-at-ns.microscopy.com} 12, 31 -- X-MS-Has-Attach: 12, 31 -- X-MS-TNEF-Correlator: 12, 31 -- Thread-Topic: [Microscopy] Scanners for TEM negatives 12, 31 -- thread-index: AceINKyAHnXgTwpbQf+BiE4T5URxGAACWnOQ 12, 31 -- From: "Bobrowski, Walter" {Walter.Bobrowski-at-pfizer.com} 12, 31 -- To: {jgross-at-neuron.uchc.edu} , {microscopy-at-microscopy.com} 12, 31 -- X-OriginalArrivalTime: 26 Apr 2007 20:05:49.0827 (UTC) FILETIME=[490CBD30:01C7883E] 12, 31 -- X-Proofpoint-Virus-Version: vendor=fsecure engine=4.65.5502:2.3.11,1.2.37,4.0.164 definitions=2007-04-26_08:2007-04-26,2007-04-26,2007-04-26 signatures=0 12, 31 -- X-Proofpoint-Spam-Reason: safe 12, 31 -- Content-Transfer-Encoding: 8bit 12, 31 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l3QK5oqQ025306 ==============================End of - Headers==============================
I have to say I agree, I have a Microtek Scanmaker 9800XL, with a TMA 1600 and I love it. It's the second Microtek I've had, and I'm pretty pleased by the way it handles TEM negs, once I figured out the best settings to use with their Silverfast software. Now I love the way it works.
I do use cut up strips of the neg holders that come with it to tape my negs across, not directly on the glass face of this scanner, and I seldom have problems with the "swirls".
Lou Ann
}
-- ~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 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
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 } ---------------------------------------------------------------------------- } } Microtek i900. Hands down. No questions asked! Great intuitive interface. Been a Microtek fan for years. } Bet regards, } Walt } }
==============================Original Headers============================== 14, 19 -- From lamiller-at-uiuc.edu Thu Apr 26 15:47:53 2007 14, 19 -- Received: from expredir4.cites.uiuc.edu (expredir4.cites.uiuc.edu [128.174.5.187]) 14, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3QKlrXp005360 14, 19 -- for {microscopy-at-microscopy.com} ; Thu, 26 Apr 2007 15:47:53 -0500 14, 19 -- Received: from redkite.cvm.uiuc.edu (redkite.cvm.uiuc.edu [130.126.18.157]) 14, 19 -- by expredir4.cites.uiuc.edu (8.13.8/8.13.8) with ESMTP id l3QKlqGX018343; 14, 19 -- Thu, 26 Apr 2007 15:47:52 -0500 (CDT) 14, 19 -- Message-ID: {46310FF8.7080702-at-uiuc.edu} 14, 19 -- Date: Thu, 26 Apr 2007 15:47:52 -0500 14, 19 -- From: Lou Ann Miller {lamiller-at-uiuc.edu} 14, 19 -- Reply-To: lamiller-at-uiuc.edu 14, 19 -- User-Agent: Thunderbird 2.0.0.0 (Macintosh/20070326) 14, 19 -- MIME-Version: 1.0 14, 19 -- To: Walter.Bobrowski-at-pfizer.com, microscopy-at-microscopy.com 14, 19 -- Subject: Re: [Microscopy] RE: Scanners for TEM negatives 14, 19 -- References: {200704262006.l3QK6Zn8026352-at-ns.microscopy.com} 14, 19 -- In-Reply-To: {200704262006.l3QK6Zn8026352-at-ns.microscopy.com} 14, 19 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 14, 19 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
Dear Michael Shaffer, If what you mean by high magnification could extend to 10Kx and beyond, then our 292UTC and 150-2DUTC traceable calibration specimens may be suitable for you. The first one is a 1-dimensional pattern (lines and spaces) with pitch 292 nm. The second one is a 2-dimensional pattern (square grid of bumps) with 144 nm pitch. Both are available in non-traceable versions at reduced cost.
Further information is at www.asmicro.com/Supplies/Calibrator_guide.htm, where you will find information about our full line of sub-micron calibration specimens.
I hope this information is helpful.
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: michael-at-shaffer.net To: donc-at-asmicro.com Sent: Wednesday, April 25, 2007 9:15 AM Subject: [a] [Microscopy] SEM standards for imaging
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Hello MSA'rs :o)
I need an imaging standard that would allow me to check the magnification and distortions present for our SEM. A good example is the set of SIRA gratings, but I thought I'd put it to the list for finding the most versatile reference standard relative to cost. My needs are (1) to check magnification in x & y for high and low mag, (2) to check orthogonality at high and low mag, and (3) to check for barrel, pin-cushioning, or trapazoidal distortions, at high and low mag ... Where high and low mag are } 2000x and {500x.
Tia & cheerios from "the rock" :o) michael shaffer
SEM/MLA Research Coordinator {http://www.mun.ca/creait/maf/} Inco Innovation Centre Memorial University St. John's, Newfoundland
==============================Original Headers============================== 5, 18 -- From michael-at-shaffer.net Wed Apr 25 08:10:15 2007 5, 18 -- Received: from n034.sc1.he.tucows.com (smtpout1476.sc1.he.tucows.com [64.97.157.176]) 5, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3PDAFp3016132 5, 18 -- for {Microscopy-at-microscopy.com} ; Wed, 25 Apr 2007 08:10:15 -0500 5, 18 -- Received: from roamingwolf (134.153.130.141) by n034.sc1.he.tucows.com (7.2.069.1) (authenticated as michael-at-shaffer.net) 5, 18 -- id 4626955E00122F30 for Microscopy-at-microscopy.com; Wed, 25 Apr 2007 13:10:10 +0000 5, 18 -- From: "michael shaffer" {michael-at-shaffer.net} 5, 18 -- To: "MSA Microscopy list" {Microscopy-at-microscopy.com} 5, 18 -- Subject: SEM standards for imaging 5, 18 -- Date: Wed, 25 Apr 2007 10:40:08 -0230 5, 18 -- Message-ID: {000201c7873b$0d874ea0$8d829986-at-CREAIT.MUN.CA} 5, 18 -- MIME-Version: 1.0 5, 18 -- Content-Type: text/plain; 5, 18 -- charset="us-ascii" 5, 18 -- Content-Transfer-Encoding: 7bit 5, 18 -- X-Mailer: Microsoft Office Outlook 11 5, 18 -- X-MIMEOLE: Produced By Microsoft MimeOLE V6.00.2900.3028 5, 18 -- Thread-index: AceHOwxUAS+JgyTXQ/2d8DZqc+Y41A== ==============================End of - Headers==============================
==============================Original Headers============================== 18, 22 -- From donc-at-asmicro.com Thu Apr 26 17:15:08 2007 18, 22 -- Received: from smtp111.sbc.mail.re2.yahoo.com (smtp111.sbc.mail.re2.yahoo.com [68.142.229.94]) 18, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id l3QMF8wl018356 18, 22 -- for {microscopy-at-microscopy.com} ; Thu, 26 Apr 2007 17:15:08 -0500 18, 22 -- Received: (qmail 90509 invoked from network); 26 Apr 2007 22:15:07 -0000 18, 22 -- Received: from unknown (HELO ASM11) (asmicro-at-sbcglobal.net-at-68.253.41.189 with login) 18, 22 -- by smtp111.sbc.mail.re2.yahoo.com with SMTP; 26 Apr 2007 22:15:07 -0000 18, 22 -- X-YMail-OSG: k2c68oYVM1kvbh2hnn8SBzOLrRl2rjiGdN8gDtMoLpCeq7v7gdOpC7J5GOD8CgjwVlYXaZW8LqTx2HK1P1iF4489.Lg3fEW_NjvXAzsO9hUXqtJ529RTL4g0kj7W6OTEhUX3q6F2bq4acDk2NLaOUFf_wfI- 18, 22 -- Message-ID: {006101c78850$221b6390$6401a8c0-at-ASM11} 18, 22 -- From: "Don Chernoff at ASM" {donc-at-asmicro.com} 18, 22 -- To: {michael-at-shaffer.net} , "Microscopy List" {microscopy-at-microscopy.com} 18, 22 -- References: {200704251315.l3PDFFF8021437-at-ns.microscopy.com} 18, 22 -- Subject: Re: [a] [Microscopy] SEM standards for imaging 18, 22 -- Date: Thu, 26 Apr 2007 18:12:40 -0400 18, 22 -- MIME-Version: 1.0 18, 22 -- Content-Type: text/plain; 18, 22 -- charset="iso-8859-1" 18, 22 -- Content-Transfer-Encoding: 7bit 18, 22 -- X-Priority: 3 18, 22 -- X-MSMail-Priority: Normal 18, 22 -- X-Mailer: Microsoft Outlook Express 6.00.2800.1807 18, 22 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2800.1896 ==============================End of - Headers==============================
We have the Microtek ScanMaker i900. The glassless holders are indeed a nice feature. I don't know what size negatives you generate, but I place the 3.25"x4" negatives "sideways" in the 4"x5" carrier and it works very well. I scan negatives at 600dpi or 1200dpi. While it does a very nice job, it is slow - takes just about 5 min to scan a single negative at the 600dpi setting. I use the USB connection; I tried the FireWire interface and couldn't get it to work, but put no time into it. The USB connection does not seem to be the limiting factor. It uses a "ScanServer" driver to interface with the device so you can switch it on and off freely without the "Eject USB device" protocol. I do have small problems at startup sometimes - errors between Adobe Photoshop and the plugin/device, but powering off the scanner and restarting the ScanServer fixes it, and once functioning it works w/o error.
When you power on the scanner it goes through some initialization that lasts ~ 2min; be patient until it is really ready - all noises really done and the indicator LEDS are steady - or it will hang.
Just to make sure I don't have to adjust for each negative to fit an 8-bit range, I scan at 16-bit with the density range set to 3.0, and then scale to the actual range and convert to 8-bit when it looks nice on the monitor for general use, keeping the original scans if warranted - they are 32Mb as TIFFs....
It does get to be a time-consuming pain scanning a rack of negatives. I keep a timer set for 5 min and change the negative when done. You can load 2 negatives at a time in the 4"x5" holder and set it up as 2 jobs, giving you 10 min break, but that is more than I can keep straight most days... I sometimes throw 6 negatives at a time on the glass transparency platen and do the equivalent of a "contact sheet", giving an even longer interval between feeding loads, but at the peril of glass platen issues.....
So overall I like it. It seems good and well made for the cost; speed is the only thing I would change. If anyone knows that I'm doing something wrong - making it slow - PLEASE tell me!
Hope this helps. Drive up and scan a few if you'd like.
Dale Callaham Central Microscopy Facility University of Massachusetts, Amherst. http://www.bio.umass.edu/microscopy
jgross-at-neuron.uchc.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 } ---------------------------------------------------------------------------- } } Our lab needs to purchase a new scanner for our TEM negatives. We } have narrowed our choices to either the Epson Perfection V750 } (recommended by Keith Young in the May 2006 issue of Microscopy } Today) or the Microtek ScanMaker i900. The major difference between } the two seems to be the "glassless" scanning feature of the Microtek. } Has anyone had first-hand experience of the film scanning ability of } either one of these scanners? Or are there any quirks that would } make one of them less desirable? } } Thank you, } Julie Gross } Kent Morest Lab } Dept. of Neuroscience } UCONN Health Center } Farmington, CT 06030 } jgross-at-neuron.uchc.edu } } } ==============================Original Headers============================== } 3, 18 -- From jgross-at-neuron.uchc.edu Thu Apr 26 13:50:49 2007 } 3, 18 -- Received: from uchc.edu (itiron1.uchc.edu [155.37.201.25]) } 3, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3QIomws020028 } 3, 18 -- for {microscopy-at-microscopy.com} ; Thu, 26 Apr 2007 13:50:49 -0500 } 3, 18 -- Received: from ([155.37.91.71]) } 3, 18 -- by itiron1.uchc.edu with ESMTP id 5502373.19449977; } 3, 18 -- Thu, 26 Apr 2007 14:50:39 -0400 } 3, 18 -- Received: from morgpc.neuron.uchc.edu ([155.37.1.96]) by itnwa.uchc.edu with Microsoft SMTPSVC(5.0.2195.6713); } 3, 18 -- Thu, 26 Apr 2007 14:50:38 -0400 } 3, 18 -- X-Mailer: QUALCOMM Windows Eudora Version 7.1.0.9 } 3, 18 -- Date: Thu, 26 Apr 2007 14:50:38 -0400 } 3, 18 -- To: microscopy-at-microscopy.com } 3, 18 -- From: Julie Gross {jgross-at-neuron.uchc.edu} } 3, 18 -- Subject: Scanners for TEM negatives } 3, 18 -- Mime-Version: 1.0 } 3, 18 -- Content-Type: text/plain; charset="us-ascii"; format=flowed } 3, 18 -- Message-ID: {ITNWA6PYzDabigPasSm00000115-at-itnwa.uchc.edu} } 3, 18 -- X-OriginalArrivalTime: 26 Apr 2007 18:50:39.0083 (UTC) FILETIME=[C86FD3B0:01C78833] } ==============================End of - Headers==============================
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Organization: San Joaquin Delta College - Electron Microscopy Program
Title-Subject: [Filtered] Dehydration of molds
Question: I am trying to figure out the best way to dehydrate and dry two types of mold, Rhizopus and A. Niger. I am going to osmium vapour fix them overnight but I would like to know if any of you have any suggestions for the dehydration and drying procedures. I am worried since the samples are so delicate. I am going to be viewing both samples using the SEM.
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Email: swtkeller-at-yahoo.com Name: Sandra Keller
Organization: TA-SICCO
Title-Subject: [Filtered] TEM-3rd party service providers
Question: Hi: I know this topic comes up on occasion, but I am wanting to tap the extremely valuable resource of this community.
I am looking for the most up-to-date information when it comes to 3rd party service companies. Companies that offer on demand service for JEOL, FEI (Philips), Hitachi, and Zeiss TEM's or any now defunct brands in the US. I am especially interested in information on organizations that offer maintenance (service) contracts, especially for JEOL TEM's.
I have also heard of some type of insurance companies providing some type of similar arrangement, but that was several years ago and I have personally not run across any organization providing this type of service. Any help on this topic would be greatly appreciated.
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Title-Subject: [Filtered] gas injection/heating TEM holder
Question: Hi, All: We are looking into buying a gas injection/heating TEM holder. It would be very grateful if you could give some suggestions and recommendation. Zhiqiang
Yes...a difficult specimen situation. If you are quick, you do not need to do any fixation.
I did this with Zeiss Supra 55VP at 200V EHT. This was with the in-lens detector. No coating.
This has to be done fast since the clusters will implode when subjected to high vacuum. VP is not all that great. Do HV and quickly.
gary g.
At 05:46 PM 4/26/2007, you wrote: } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
One technique I have used successfully in the past is fixing with osmium vapour, dehydrating using vapor diffusion dehydration and then critical point drying. This can preserve the conidial structure of Aspergillus very well. The original reference is:
EJ King and MF Brown 1983 A technique for preserving aerial fungal structures for scanning electron microscopy. Canadian Journal of Microbiology 29:653-658
Ian
Ian Hallett Sensory and Consumer Science - Microscopy HortResearch, Mt Albert Research Centre Private Bag 92 169, Auckland Mail Centre Auckland 1142, New Zealand +64-9-815 4200 ext 7002
-----Original Message----- X-from: arollins-at-hotmail.com [mailto:arollins-at-hotmail.com] Sent: Friday, 27 April 2007 1:48 p.m. To: Ian Hallett
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Organization: San Joaquin Delta College - Electron Microscopy Program
Title-Subject: [Filtered] Dehydration of molds
Question: I am trying to figure out the best way to dehydrate and dry two types of mold, Rhizopus and A. Niger. I am going to osmium vapour fix them overnight but I would like to know if any of you have any suggestions for the dehydration and drying procedures. I am worried since the samples are so delicate. I am going to be viewing both samples using the SEM.
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==============================Original Headers============================== 21, 30 -- From IHallett-at-hortresearch.co.nz Thu Apr 26 22:21:24 2007 21, 30 -- Received: from hortresearch.co.nz (mscan.hortresearch.co.nz [202.36.134.15]) 21, 30 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3R3LN7K026953 21, 30 -- for {microscopy-at-microscopy.com} ; Thu, 26 Apr 2007 22:21:24 -0500 21, 30 -- Received: from aklexf01.hort.net.nz ([10.16.1.14]) by hortresearch.co.nz 21, 30 -- with HortResearch; Fri, 27 Apr 2007 15:37:12 +1200 21, 30 -- Received: from AKLEXB01.hort.net.nz ([10.16.1.15]) by aklexf01.hort.net.nz 21, 30 -- with Microsoft SMTPSVC(6.0.3790.1830); Fri, 27 Apr 2007 15:21:21 +1200 21, 30 -- X-MimeOLE: Produced By Microsoft Exchange V6.5.7226.0 21, 30 -- Content-class: urn:content-classes:message 21, 30 -- MIME-Version: 1.0 21, 30 -- Content-Type: text/plain; 21, 30 -- charset=us-ascii 21, 30 -- Subject: RE: [Microscopy] viaWWW: Dehydration of molds 21, 30 -- Date: Fri, 27 Apr 2007 15:21:20 +1200 21, 30 -- Message-ID: {D3BAD63C088F3C48AEB385E881359F870305B1DD-at-AKLEXB01.hort.net.nz} 21, 30 -- In-Reply-To: {200704270147.l3R1lY2d023281-at-ns.microscopy.com} 21, 30 -- X-MS-Has-Attach: 21, 30 -- X-MS-TNEF-Correlator: 21, 30 -- Thread-Topic: [Microscopy] viaWWW: Dehydration of molds 21, 30 -- Thread-Index: AceIbghcHu4/JvluRkad/8RipC6ApgADG3lg 21, 30 -- From: "Ian Hallett" {IHallett-at-hortresearch.co.nz} 21, 30 -- To: {arollins-at-hotmail.com} , {microscopy-at-microscopy.com} 21, 30 -- X-OriginalArrivalTime: 27 Apr 2007 03:21:21.0025 (UTC) 21, 30 -- FILETIME=[20751F10:01C7887B] 21, 30 -- X-imss-version: 2.046 21, 30 -- X-imss-result: Passed 21, 30 -- X-imss-approveListMatch: *-at-hortresearch.co.nz 21, 30 -- Content-Transfer-Encoding: 8bit 21, 30 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l3R3LN7K026953 ==============================End of - Headers==============================
I don't know if I'm Keith Young, but to be honest Microtek haven't bought out a new top flight scanner for years and their ArtixScan designs are living on their old name and hardware. Agfa scanners were simply rebadged Microtek's until they withdrew from the scanner market along with Fuji. The Microtek ArtixScan 2500f is pretty much the £5,000 Agfa Duoscan 2550T from 7 years ago, now with firewire instead of SCSI. Not that these professional Microtek scanners are rubbish, just a bit expensive for a similar scan quality as the Epson 4990 Photo (and their scans are probably a bit inferior in resolution to the 6,400dpi Epson V750 - the Agfa 2550T and Epson 4990 are). As I mentioned the upmarket Microtek scanners may have a far better build quality though. The cheaper Microtek i900, introduced I think in 2004, has a resolution of about 3,200 dpi. It's glassless design could help reduce problems with dust, although the scanner optics is no longer 'sealed behind glass'. It uses 4x5" film holders, a 8x10" glass sandwich or the A4 top bed for reflective scans. It has been reported as very slow, but then most prosumer flatbeds at this price aren't that fast. I doubt if you would be disappointed if you bought the Microtek i900 or the Epson V750 (or the Epson V700 or 4990 Photo for that matter).
The only way to find out if, at 3,200 dpi quoted resolution, the i900 produces better scans (& faster) than the 6,400 dpi quoted Epson 750 is to scan the same negative in both, at the same and maximum resolution (I've never tried the i900, although our Microtek/Agfa Duoscan 2550T looks very similar). There was also the similar film only Epson F3200 [dpi], that could scan negatives greater than 9 x 6 cm using 4x5" film holders, but it seems to have been withdrawn recently.
I did own an old Nikon 2,700 dpi dedicated 35mm slide scanner at home but it simply took up too much room and it's scan quality was worse than the Canon 9950F I got to replace it, particularly for colour negatives - plus the Canon only does reflective scans to A4, but annoyingly Canon's Scangear restricts film sizes to it's film holders. In comparison the V700/V750 pro can go to A4 for film, with film in holders or just lying on the platen. I have only used a 6,400 dpi V750 Pro briefly for 35mm colour slide film (my main interest), and its scan quality was a bit better than my Canon 9950F, i.e. the fuzz was a tiny tiny bit sharper, it's not streets ahead at all (although it's Epson Scan twain interface was in another league). Plus the 35mm slide holder retaining clips on the V750 are rather flimsy (although the Canon and Epson 4990 don't even bother with retaining clips, using gravity to hold them in place). I don't know how good the 35mm clips on the i900 are (they don't look great either). At work our 4,800 dpi Epson 4990 Photo is more than adequate for 1,200 dpi B&W TEM film scans, so I haven’t bothered upgrading to a V750 Pro. The V750 does also offer an optional fluid mount scan accessory like every expensive drum scanners, but that’s unlikely to be of interest for TEM film scanning. It's meant to get rid of Newton rings (swirls) and can assist in hiding scratches and dust, but it often introduces bubbles instead.
However you will get superior scans from an £8,000 Hasselblad 8,000 dpi [Imacon] 848 semi-drum scanner, but often it will be the film grain that looks a bit better resolved rather than actual image detail. Plus you must use the film holders - it scans fast though. It also has no glass between the film and detector. Things like FARE/ICE dust removal and accurate colour reproduction (the latter being essential for professionals that need correct skin tones or accurate reproductions of museum articles) are irrelevant for your B&W scientific images.
The excellent 4000 dpi 35mm Nikon Coolscan 9000 [3-CCD sensor] film scanner comes in at £2,000. However the Nikon 9000 can only scan film up to 9 x 6 cm in size and histological sections on glass slides (too small for most TEM negatives unless you cut them down). Plus it has a lot of software/hardware for 35mm colour film thrown in you probably won't need.
To be honest the 4,800 dpi Epson 4990 Photo at £250 is more than adequate for large format negatives (TEM film) as it can go up to A4 film (six TEM negatives per scan) and Epson's Scan is an easier Twain interface than Silverfast and Canons awful Scangear (there is also the highly regarded Vuescan third party twain software available).
In practice it is most unlikely that will often want to scan at more than 1,200 dpi anyway as the resulting file size will be enormous and highly prone to file corruption on the PC (but images should have noticeably better resolution if scanned at 2,400dpi). Most scan just for publication or for PC on-screen image analysis, rather than printing to A4/A3. A Epson 4990 Photo 4,800 dpi scan of Kodak 4489 negative is around 250 Mb in TIF format, at 2,400 dpi it drops to a 60Mb TIF file with little different in image quality. There no reason however not to go for the better 6,400 dpi Epson V750 Pro, even if you scan mainly at 1,200 dpi, as it's only a bit more money (and not ours). The 6,400 dpi will be useful for enlargement of selected areas (scanning at 3,200 dpi being optimal).
Perhaps one downside to the V750 Pro is that it works best with the slide/film holders and their little leg supports. Some sizes of TEM film may have small parts of the image hidden by a 4x5" film holder. You can source third party film holders though (see Doug Fisher), or you can just drop them onto the platen and live with a possible small drop in resolution. As mentioned you can get interference patterns (swirls) if the negative is placed on the platen rather than using the 4x5" film holder. This is probably caused by a slightly damp negative (high humidity) or the glass platen having greasy finger marks. So far I have always got rid of this by cleaning the platen (50% propanol/50% water with soft tissue and air-bulb blow dry - also great for finger smudged VDU screens). Remove film from the platen with a sheet of card rather than fingers (i.e. never touch the glass platen). No-one here ever bothers to use the Epson 4990's 4x5" film holders as it obscures part of the negative, only takes a few films and is far more fiddly. They just place six TEM negatives directly on the platen (correct side up).
If you are really really serious about scanning film for some reason there is also the likes of professional flatbed scanners such as the Kodak ‘Creo’£15k [10,000 dpi] IQSmart 3, the £30k [8000 to 14000 dpi] EverSmart Supreme II flatbeds (www.kodak.com). However cheaper consumer ‘photo’ flat beds such as the new 6,400 dpi Epson V750 and V750 Pro come in at £500 and £650 respectively, and I can't see any reason to spend more for infrequent scans of large format B&W film, as you will rarely, if ever, scan at maximum resolution. They also have convenient USB2 and Firewire interfaces and work with Apples or PCs.
You will have to spend a bit of time working on the best twain scan settings and images will probably need some post-editing in Photoshop CS3 to get optimal scanned image quality.
But the choice is yours. Do however have a look at http://www.photo-i.co.uk for Independent reviews of some prosumer large format (A4) film scanners (not the i900). Also note that these cheaper scanners are for light use, if you want to scan hundreds of films a day, and not be chained to the scanner, look elsewhere and with a deeper pocket.
Regards
Keith
PS. My 'Scanning film on a budget' Microscopy Today article has been updated and reprinted in Infocus (March 2007) the UK Royal Microscopical society's magazine. I can't send a pdf though as the editor hasn't passed on a corrected version yet. I can only send a scan of the article taken using my home 4,800 dpi Canon 9950F (scanned at 600 dpi) - but don't try that at home with your £2,000 Nikon Coolscan 9000.
PPS. As mentioned the now withdrawn Canon 9550F has a terrible twain software interface(Scangear), the Epsons Scan one is miles better, so I would avoid the Canon even if you can still find it [unless its only ever being used for scanning photographs or with standard film holder sizes]. Interestingly Canon scrapped their dedicated 35mm film scanners for the 9950F (and they should know something about film photography) but the poor Scangear twain interface didn't make many friends. I bought the 9950F just before the Epson 4990 Photo came in - which I later purchased for work use, mainly for TEM negatives, 2D gel film and reflective scans.
--------------------------- Dr Keith J Morris Imaging Facilities Manager Cell Biology Institute of Ophthalmology UCL, London EC1V 9EL
-----Original Message----- X-from: jgross-at-neuron.uchc.edu [mailto:jgross-at-neuron.uchc.edu] Sent: 26 April 2007 19:55 To: keith.morris-at-ucl.ac.uk
Our lab needs to purchase a new scanner for our TEM negatives. We have narrowed our choices to either the Epson Perfection V750 (recommended by Keith Young in the May 2006 issue of Microscopy Today) or the Microtek ScanMaker i900. The major difference between the two seems to be the "glassless" scanning feature of the Microtek. Has anyone had first-hand experience of the film scanning ability of either one of these scanners? Or are there any quirks that would make one of them less desirable?
Thank you, Julie Gross Kent Morest Lab Dept. of Neuroscience UCONN Health Center Farmington, CT 06030 jgross-at-neuron.uchc.edu
==============================Original Headers============================== 32, 24 -- From keith.morris-at-ucl.ac.uk Fri Apr 27 07:11:58 2007 32, 24 -- Received: from smtp1.global.net.uk (smtp1.global.net.uk [80.189.94.53]) 32, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3RCBvDs015635 32, 24 -- for {microscopy-at-microscopy.com} ; Fri, 27 Apr 2007 07:11:57 -0500 32, 24 -- Received: from 141.11.rb4.adsl.brightview.com ([80.189.11.141] helo=loungepc) 32, 24 -- by smtp1.global.net.uk with esmtp (Exim 4.42) 32, 24 -- id 1HhPIt-000KbJ-3b 32, 24 -- for microscopy-at-microscopy.com; Fri, 27 Apr 2007 13:11:59 +0100 32, 24 -- From: "Keith Morris" {keith.morris-at-ucl.ac.uk} 32, 24 -- To: {microscopy-at-microscopy.com} 32, 24 -- References: {200704261855.l3QItI2m026586-at-ns.microscopy.com} 32, 24 -- Subject: RE: [Microscopy] Scanners for TEM negatives 32, 24 -- Date: Fri, 27 Apr 2007 13:11:57 +0100 32, 24 -- Message-ID: {000901c788c5$40acf4c0$0201a8c0-at-loungepc} 32, 24 -- MIME-Version: 1.0 32, 24 -- Content-Type: text/plain; 32, 24 -- charset="iso-8859-1" 32, 24 -- X-Mailer: Microsoft Office Outlook 11 32, 24 -- Thread-Index: AceINHLb4TXSJai/QcGWrztdBBdnGgAHZTog 32, 24 -- In-Reply-To: {200704261855.l3QItI2m026586-at-ns.microscopy.com} 32, 24 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.3028 32, 24 -- Authenticated-Sender: 32, 24 -- Content-Transfer-Encoding: 8bit 32, 24 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l3RCBvDs015635 ==============================End of - Headers==============================
What an interesting idea! I like MM, oh wait, Disney has that locked up!
While we are on the subject, can we arrange to get people to stop using "electron microscope" when they mean transmission electron microscope or scanning electron microscope?
Oh!, If I was king of the world................
edelmare-at-muohio.e du To: frank.karl-at-degussa.com cc: 04/26/2007 03:54 Subject: [Microscopy] Re: Terminology - please vote! PM Please respond to edelmare
The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
O.k. it sounds fairly unamimous that "Electron Microprobe / EPMA" is the genrally used accepted terminology for a "WDS-Microprobe" and is better than simply microprobe.
BUT Phil hits the nail on the head with regards to "mechanical microprobe" type devices.
==} "Let's start using [a term] before some company trademarks it so no one else can use it."
So what shall it be folks? "Mechanical Microprobe"? "Nanomanipulator"? Something else?
Let us the microscopists decide before someone trademarks it.
} } I have to agree with Warren, especially perhaps his last paragraph. } "Microprobe" has a definite meaning to me and most microscopists. } I wouldn't suggest using "nanoprobe", as that doesn't really solve } the problem of confusing terms. Perhaps a simple addition of an } adjective to make it a noun phrase, like "mechanical microprobe". } Micromanipulator is a well-known term, so the next step would be } nanomanipulator. Let's start using that before some company } trademarks it so no one else can use it. } } Phil } } } I still associate microprobe with the scanning electron beam device } } equipped with wavelength spectrometers for the express purpose of x-ray } } microanalysis. } } } } I refer to the little thing used to hold and move objects around as } } micro-manipulators. } } } } I will admit that micro-probe can refer to a lot of things. I wouldn't } } be surprised if the microelectronics folks adopted the term to refer to } } a microscopic test lead. I also suppose the scanning probe microscope } } folks could refer to their tips as microprobes. But to me, a microprobe } } will be those things like ARL used to make. } } } } Maybe that makes me old school, but it seems like a decent working } } definition. I am getting old enough people probably expect me to be a } } bit pig-headed and crotchety anyway. I might as well live up to it. {g} } } } } Warren Straszheim } } } } } } -----Original Message----- } } X-from: edelmare-at-muohio.edu [mailto:edelmare-at-muohio.edu] } } Sent: Wednesday, April 25, 2007 3:44 PM } } To: wesaia-at-iastate.edu } } Subject: [Microscopy] Question of Terminology - "Microprobe" } } } } } } O.k., folks Lets see if the Microscopy and Microanalysis community } } can come an agreement. } } } } Traditionally the term "Microprobe" has been used to refer to a } } dedicated wave length dispersive x-ray microanalysis } } microscope/instrument (XWDS or WDS). Is this still a valid use for } } the the term "Microprobe"? Or is it antiquated? } } } } . . . because it comes into conflict a "newer" use of the word } } "microprobe" as refering to a "small" physical probing device, } } typically mechanical, for manipulating or testing small items. Would } } these probing-manipuating-testing devices be better called } } "Nanoprobes" (even thought they may actually handle items / objects } } in the micrometer to 100's-micrometer range). } } } } Context will NOT sufice to distinguish between the two as } } microscopists now deal with both. } } } } Anyone care to share comments or opinions ? } } } } } } Richard E. Edelmann, Ph.D. } } EXPO Editor, Microscopy and Microanalysis Supplement } } Electron Microscopy Facility Director } } 364 Pearson Hall } } Miami University, Oxford, OH 45056 } } Ph: 513.529.5712 Fax: 513.529.4243 } } E-mail: edelmare-at-muohio.edu } } http://www.emf.muohio.edu } -- } 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============================== } 3, 22 -- From oshel1pe-at-cmich.edu Thu Apr 26 07:56:59 2007 } 3, 22 -- Received: from ob2.cmich.edu (ob2.cmich.edu [141.209.20.21]) } 3, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3QCuxCm010367 } 3, 22 -- for {Microscopy-at-microscopy.com} ; Thu, 26 Apr 2007 07:56:59 -0500 } 3, 22 -- Received: from egatea.central.cmich.local ([141.209.15.74]) } 3, 22 -- by ob2.cmich.edu (8.12.10/8.12.10) with ESMTP id l3QDL26h026847 } 3, 22 -- for {Microscopy-at-microscopy.com} ; Thu, 26 Apr 2007 09:21:02 -0400 } 3, 22 -- Received: from [141.209.160.249] ([141.209.160.249]) by egatea.central.cmich.local with Microsoft SMTPSVC(6.0.3790.1830); } 3, 22 -- Thu, 26 Apr 2007 08:56:56 -0400 } 3, 22 -- Mime-Version: 1.0 } 3, 22 -- Message-Id: {f06230904c25650f2b54c-at-[141.209.160.249]} } 3, 22 -- In-Reply-To: {200704252113.l3PLDaRo014950-at-ns.microscopy.com} } 3, 22 -- References: {200704252113.l3PLDaRo014950-at-ns.microscopy.com} } 3, 22 -- Date: Thu, 26 Apr 2007 08:56:55 -0400 } 3, 22 -- To: Microscopy-at-microscopy.com } 3, 22 -- From: Philip Oshel {oshel1pe-at-cmich.edu} } 3, 22 -- Subject: [Microscopy] RE: Question of Terminology - "Microprobe" } 3, 22 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" } 3, 22 -- X-OriginalArrivalTime: 26 Apr 2007 12:56:56.0194 (UTC) FILETIME=[5E9BF620:01C78802] } 3, 22 -- X-CanItPRO-Stream: default } 3, 22 -- X-Spam-Score: -3.7 () L_EXCH_MF,MY_HTML_OBFU } 3, 22 -- X-Scanned-By: CanIt (www . roaringpenguin . com) on 141.209.20.21 } ==============================End of - Headers==============================
Richard E. Edelmann, Ph.D. EXPO Editor, Microscopy and Microanalysis Supplement Electron Microscopy Facility Director 364 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu http://www.emf.muohio.edu
==============================Original Headers============================== 9, 24 -- From edelmare-at-muohio.edu Thu Apr 26 14:53:04 2007 9, 24 -- Received: from mulnx12.mcs.muohio.edu (mulnx12.mcs.muohio.edu [134.53.6.67]) 9, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3QJr371002286 9, 24 -- for {microscopy-at-Microscopy.com} ; Thu, 26 Apr 2007 14:53:04 -0500 9, 24 -- Received: from mulnx23.mcs.muohio.edu (mulnx23.mcs.muohio.edu [134.53.6.10]) 9, 24 -- by mulnx12.mcs.muohio.edu (Switch-3.1.8/Switch-3.1.7) with ESMTP id l3QJr2IP013948 9, 24 -- for {microscopy-at-Microscopy.com} ; Thu, 26 Apr 2007 15:53:02 -0400 9, 24 -- Received: from [192.168.1.23] ([134.53.14.105]) 9, 24 -- by mulnx23.mcs.muohio.edu (Switch-3.1.8/Switch-3.1.7) with ESMTP id l3QJr25X002076 9, 24 -- for {microscopy-at-Microscopy.com} ; Thu, 26 Apr 2007 15:53:02 -0400 9, 24 -- From: "Richard E. Edelmann" {edelmare-at-muohio.edu} 9, 24 -- To: microscopy-at-Microscopy.com 9, 24 -- Date: Thu, 26 Apr 2007 15:53:02 -0400 9, 24 -- MIME-Version: 1.0 9, 24 -- Subject: Re: Terminology - please vote! 9, 24 -- Message-ID: {4630CADE.22716.27D1525-at-edelmare.muohio.edu} 9, 24 -- Priority: normal 9, 24 -- In-reply-to: {200704261258.l3QCw7lS011622-at-ns.microscopy.com} 9, 24 -- References: {200704261258.l3QCw7lS011622-at-ns.microscopy.com} 9, 24 -- X-mailer: Pegasus Mail for Windows (4.41) 9, 24 -- Content-type: text/plain; charset=US-ASCII 9, 24 -- Content-transfer-encoding: 7BIT 9, 24 -- Content-description: Mail message body 9, 24 -- X-Scanned-By: MIMEDefang 2.57 on 134.53.6.67 ==============================End of - Headers==============================
==============================Original Headers============================== 28, 18 -- From frank.karl-at-degussa.com Fri Apr 27 07:32:24 2007 28, 18 -- Received: from framailout1.rz.itson.com (mailout2.degussa.com [149.216.91.173]) 28, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3RCWNFo027330 28, 18 -- for {microscopy-at-msa.microscopy.com} ; Fri, 27 Apr 2007 07:32:23 -0500 28, 18 -- Received: from mobuscomm01.mail.degussa.com (uss1026.applications.degussanet.com [10.88.88.98]) 28, 18 -- by framailout1.rz.itson.com (8.13.4/8.13.4/Debian-3sarge3) with ESMTP id l3RCWK3Z018687; 28, 18 -- Fri, 27 Apr 2007 14:32:20 +0200 28, 18 -- In-Reply-To: {200704261954.l3QJsJJB004380-at-ns.microscopy.com} 28, 18 -- Subject: Re: [Microscopy] Re: Terminology - please vote! 28, 18 -- To: edelmare-at-muohio.edu, microscopy-at-msa.microscopy.com 28, 18 -- X-Mailer: Lotus Notes Release 6.5.5 November 30, 2005 28, 18 -- Message-ID: {OF0DF86015.A8E58261-ON862572CA.004470E9-852572CA.0044DC6B-at-degussa.com} 28, 18 -- From: frank.karl-at-degussa.com 28, 18 -- Date: Fri, 27 Apr 2007 08:32:16 -0400 28, 18 -- X-MIMETrack: Serialize by Router on MOBUSComm01/DHexternal/US(Release 6.5.1|January 21, 2004) at 28, 18 -- 04/27/2007 07:32:21 AM 28, 18 -- MIME-Version: 1.0 28, 18 -- Content-type: text/plain; charset=US-ASCII ==============================End of - Headers==============================
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==============================Original Headers============================== 7, 27 -- From eschumacher-at-mccrone.com Fri Apr 27 08:00:55 2007 7, 27 -- Received: from pgp.mccrone.com (McCrone-Associates-1051626.cust-rtr.ameritech.net [68.23.63.122] (may be forged)) 7, 27 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3RD0tQV018538 7, 27 -- for {Microscopy-at-MSA.Microscopy.Com} ; Fri, 27 Apr 2007 08:00:55 -0500 7, 27 -- Received: from pgp.mccrone.com (localhost [127.0.0.1]) 7, 27 -- by pgp.mccrone.com (Postfix) with ESMTP id 913381A800B 7, 27 -- for {Microscopy-at-MSA.Microscopy.Com} ; Fri, 27 Apr 2007 08:00:55 -0500 (CDT) 7, 27 -- Received: from MCCRONEMSG.tmg.mccrone.com ([192.168.101.20]) 7, 27 -- by pgp.mccrone.com (PGP Universal service); 7, 27 -- Fri, 27 Apr 2007 08:00:55 -0500 7, 27 -- X-PGP-Universal: processed 7, 27 -- Content-class: urn:content-classes:message 7, 27 -- MIME-Version: 1.0 7, 27 -- Content-Type: text/plain; 7, 27 -- charset="us-ascii" 7, 27 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 7, 27 -- Subject: Short Course Announcement: Optical Crystallography 7, 27 -- Date: Fri, 27 Apr 2007 08:00:44 -0500 7, 27 -- Message-ID: {2A27CE0EC2A5C748974EA513DFB285A7B97582-at-MCCRONEMSG.tmg.mccrone.com} 7, 27 -- X-MS-Has-Attach: 7, 27 -- X-MS-TNEF-Correlator: 7, 27 -- Thread-Topic: Short Course Announcement: Optical Crystallography 7, 27 -- Thread-Index: AceIzBBQLyfr9zvzQHGLLBJ2D8P4Xw== 7, 27 -- From: "Elaine F. Schumacher" {eschumacher-at-mccrone.com} 7, 27 -- To: {Microscopy-at-MSA.Microscopy.Com} 7, 27 -- Content-Transfer-Encoding: 8bit 7, 27 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l3RD0tQV018538 ==============================End of - Headers==============================
I looked in the headers of the out of offices messages are sent directly to me and the list. Nestor has no control over that. Nestor does a very good job scrubbing the list to stop them from going out to the list but he has no way to stop the first one from going back to people that post to the list.
He could take parasitics action a Yahoo does and not mail any more messages out to anyone that bounces a message for any reason back to the list but most people don't think that the right way to handle a professional list. Having to delete a half dozen emails is not going to traumatize any of us.
Even Yahoo in its Draconian ways hasn't done away this problem as you still get the messages bounce back to you the first time there is a problem with a email delivery for any reason or there is an out of office message.
Compared to other list Nestor runs a very good one that is a great service to microscopist all over the world.
Being a Monday morning quarter back on how to run a list is easy but running a list as big as this one isn't. Some of the organizations we work for require that we have the out of office messages and not access private email on the clock. So those of use that work there don't have much choice in the matter.
Gordon Couger www.science-info.net Repository of old microscope documentation
tivol-at-caltech.edu wrote: } On Apr 20, 2007, at 6:49 PM, gary-at-gaugler.com wrote: } } } } The reason NOT to post to the list is that you get } } volumes of "Out Of Office" messages. This is the } } same as when you make a new posting. Most of the } } defunct messages are trapped by Eudora but quite } } a few still get though....sigh. } } } } Some do not have this subject but say that they } } are gone from xx to yy. } } } } } Dear Gary, } Is it possible to have all OOO messages have the subject line be "Out } Of Office"? If so, Nestor could request it, but if the company that } one works for has a standard OOO heading, that may be impractical. } 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============================== 9, 20 -- From gcouger-at-science-info.net Fri Apr 27 10:40:19 2007 9, 20 -- Received: from smtp101.biz.mail.mud.yahoo.com (smtp101.biz.mail.mud.yahoo.com [68.142.200.236]) 9, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id l3RFeJDX000463 9, 20 -- for {microscopy-at-microscopy.com} ; Fri, 27 Apr 2007 10:40:19 -0500 9, 20 -- Received: (qmail 37059 invoked from network); 27 Apr 2007 15:40:18 -0000 9, 20 -- Received: from unknown (HELO ?192.168.1.100?) (gcouger-at-science-info.net-at-74.195.225.38 with plain) 9, 20 -- by smtp101.biz.mail.mud.yahoo.com with SMTP; 27 Apr 2007 15:40:18 -0000 9, 20 -- X-YMail-OSG: ENMo0JAVM1nurTJc_wB5DtX8KCVq7T154qsHGgrSvpGCLz0a7mu55.edjOrXBKjHRBqINo4LBAJgD8T0QIgH29Vu7eh6RdpRaNEN2HhxTZnturqIKN0- 9, 20 -- Message-ID: {46321967.4090305-at-science-info.net} 9, 20 -- Date: Fri, 27 Apr 2007 10:40:23 -0500 9, 20 -- From: Gordon Couger {gcouger-at-science-info.net} 9, 20 -- User-Agent: Thunderbird 1.5.0.10 (Macintosh/20070221) 9, 20 -- MIME-Version: 1.0 9, 20 -- To: tivol-at-caltech.edu, microscopy-at-microscopy.com, 9, 20 -- Gary Gaugler {gary-at-gaugler.com} 9, 20 -- Subject: Re: [Microscopy] 2nd for posting to the list 9, 20 -- References: {200704231643.l3NGhemP006693-at-ns.microscopy.com} 9, 20 -- In-Reply-To: {200704231643.l3NGhemP006693-at-ns.microscopy.com} 9, 20 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 9, 20 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
I have to agree 100% on this. When I post to this list I may get 4-6 "out of office" replies. Big whoop.
I an unendingly puzzled as to why this is such a big issue.
Randy
-----Original Message----- X-from: gcouger-at-science-info.net [mailto:gcouger-at-science-info.net] Sent: Friday, April 27, 2007 10:42 AM To: Tindall, Randy D.
Bill, Gary & the list,
I looked in the headers of the out of offices messages are sent directly to me and the list. Nestor has no control over that. Nestor does a very good job scrubbing the list to stop them from going out to the list but he has no way to stop the first one from going back to people that post to the list.
He could take parasitics action a Yahoo does and not mail any more messages out to anyone that bounces a message for any reason back to the list but most people don't think that the right way to handle a professional list. Having to delete a half dozen emails is not going to traumatize any of us.
Even Yahoo in its Draconian ways hasn't done away this problem as you still get the messages bounce back to you the first time there is a problem with a email delivery for any reason or there is an out of office message.
Compared to other list Nestor runs a very good one that is a great service to microscopist all over the world.
Being a Monday morning quarter back on how to run a list is easy but running a list as big as this one isn't. Some of the organizations we work for require that we have the out of office messages and not access private email on the clock. So those of use that work there don't have much choice in the matter.
Gordon Couger www.science-info.net Repository of old microscope documentation
tivol-at-caltech.edu wrote: } On Apr 20, 2007, at 6:49 PM, gary-at-gaugler.com wrote: } } } } The reason NOT to post to the list is that you get volumes of "Out Of
} } Office" messages. This is the same as when you make a new posting. } } Most of the defunct messages are trapped by Eudora but quite a few } } still get though....sigh. } } } } Some do not have this subject but say that they are gone from xx to } } yy. } } } } } Dear Gary, } Is it possible to have all OOO messages have the subject line be "Out } Of Office"? If so, Nestor could request it, but if the company that } one works for has a standard OOO heading, that may be impractical. } 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============================== 9, 20 -- From gcouger-at-science-info.net Fri Apr 27 10:40:19 2007 9, 20 -- Received: from smtp101.biz.mail.mud.yahoo.com (smtp101.biz.mail.mud.yahoo.com [68.142.200.236]) 9, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id l3RFeJDX000463 9, 20 -- for {microscopy-at-microscopy.com} ; Fri, 27 Apr 2007 10:40:19 -0500 9, 20 -- Received: (qmail 37059 invoked from network); 27 Apr 2007 15:40:18 -0000 9, 20 -- Received: from unknown (HELO ?192.168.1.100?) (gcouger-at-science-info.net-at-74.195.225.38 with plain) 9, 20 -- by smtp101.biz.mail.mud.yahoo.com with SMTP; 27 Apr 2007 15:40:18 -0000 9, 20 -- X-YMail-OSG: ENMo0JAVM1nurTJc_wB5DtX8KCVq7T154qsHGgrSvpGCLz0a7mu55.edjOrXBKjHRBqINo4L BAJgD8T0QIgH29Vu7eh6RdpRaNEN2HhxTZnturqIKN0- 9, 20 -- Message-ID: {46321967.4090305-at-science-info.net} 9, 20 -- Date: Fri, 27 Apr 2007 10:40:23 -0500 9, 20 -- From: Gordon Couger {gcouger-at-science-info.net} 9, 20 -- User-Agent: Thunderbird 1.5.0.10 (Macintosh/20070221) 9, 20 -- MIME-Version: 1.0 9, 20 -- To: tivol-at-caltech.edu, microscopy-at-microscopy.com, 9, 20 -- Gary Gaugler {gary-at-gaugler.com} 9, 20 -- Subject: Re: [Microscopy] 2nd for posting to the list 9, 20 -- References: {200704231643.l3NGhemP006693-at-ns.microscopy.com} 9, 20 -- In-Reply-To: {200704231643.l3NGhemP006693-at-ns.microscopy.com} 9, 20 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 9, 20 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
==============================Original Headers============================== 21, 25 -- From TindallR-at-missouri.edu Fri Apr 27 10:51:10 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 l3RFp9g4010722 21, 25 -- for {microscopy-at-microscopy.com} ; Fri, 27 Apr 2007 10:51:10 -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 -- Fri, 27 Apr 2007 10:51:09 -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: 2nd for posting to the list 21, 25 -- Date: Fri, 27 Apr 2007 10:51:08 -0500 21, 25 -- Message-ID: {91108EF9255B394CBF8B7E3789814A4101C0B987-at-UM-XMAIL08.um.umsystem.edu} 21, 25 -- In-Reply-To: {200704271542.l3RFgGVW002515-at-ns.microscopy.com} 21, 25 -- X-MS-Has-Attach: 21, 25 -- X-MS-TNEF-Correlator: 21, 25 -- Thread-Topic: [Microscopy] Re: 2nd for posting to the list 21, 25 -- Thread-Index: AceI4qKf9T5eS2R7QkSvmaMtejYvQgAAPhAw 21, 25 -- References: {200704271542.l3RFgGVW002515-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: 27 Apr 2007 15:51:09.0198 (UTC) FILETIME=[DF7F8AE0:01C788E3] 21, 25 -- Content-Transfer-Encoding: 8bit 21, 25 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l3RFp9g4010722 ==============================End of - Headers==============================
Is there any software available that will allow the calculation of pair correlation function from sets of x,y coordinates? I have a number of electron micrographs with colloidal gold staining and I would like to do an analysis of particle distribution for single and double labeling. Is there a way to do this with image J?
Thanks.
George
George P. Leser, PhD Dept. Biochemistry, Molecular Biology, and Cell Biology Northwestern University Hogan 2-100 2153 North Campus Drive Evanston, IL 60208
g-leser-at-northwestern.edu
==============================Original Headers============================== 9, 18 -- From g-leser-at-northwestern.edu Fri Apr 27 11:32:51 2007 9, 18 -- Received: from merle.it.northwestern.edu (merle.it.northwestern.edu [129.105.16.57]) 9, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3RGWoWx024508 9, 18 -- for {Microscopy-at-microscopy.com} ; Fri, 27 Apr 2007 11:32:50 -0500 9, 18 -- Received: from hal9000-3ecc03cf (dhcp-129-105-38-116.bmbcb.northwestern.edu [129.105.38.116]) 9, 18 -- by merle.it.northwestern.edu (Postfix) with ESMTP id BEF4C77BC 9, 18 -- for {Microscopy-at-microscopy.com} ; Fri, 27 Apr 2007 11:29:46 -0500 (CDT) 9, 18 -- Subject: nearest neighbor analysis 9, 18 -- Reply-To: g-leser-at-northwestern.edu 9, 18 -- Content-Type: text/plain 9, 18 -- MIME-Version: 1.0 9, 18 -- X-Mailer: Info Select 2007 (9.00.36) 9, 18 -- From: "George P. Leser" {g-leser-at-northwestern.edu} 9, 18 -- To: ListServer {Microscopy-at-microscopy.com} 9, 18 -- Date: Fri, 27 Apr 2007 11:29:53 -0500 9, 18 -- Message-Id: {20070427162946.BEF4C77BC-at-merle.it.northwestern.edu} 9, 18 -- Content-Transfer-Encoding: 8bit 9, 18 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l3RGWoWx024508 ==============================End of - Headers==============================
-- Shawn Mikula, Ph.D., Postdoctoral Scholar Center for Neuroscience University of California-Davis, 1544 Newton Court, Davis, CA 95618, Phone: 530-754-9209 Fax: 530-754-9136 mail: samikula-at-ucdavis.edu web: http://brainmaps.org
----- Original Message ----- X-from: {g-leser-at-northwestern.edu} To: {samikula-at-ucdavis.edu} Sent: Friday, April 27, 2007 9:39 AM
On Apr 27, 2007, at 5:32 AM, frank.karl-at-degussa.com wrote:
} While we are on the subject, can we arrange to get people to stop using } "electron microscope" when they mean transmission electron microscope } or } scanning electron microscope? } Dear Frank, Maybe we could restrict "electron microscope" to those instruments with both capabilities; it's better than referring to them as scanning/transmission electron microscopes. If the instruments are designed for cryo, the designation "scanning/transmission electron cryo-microscopes" is almost Germanic. 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 Fri Apr 27 12:02:38 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 l3RH2bRb012173 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Fri, 27 Apr 2007 12:02:38 -0500 4, 22 -- Received: from fire-dog.its.caltech.edu (fire-dog [192.168.1.4]) 4, 22 -- by fire-ox-postvirus (Postfix) with ESMTP id B2E323557B 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Fri, 27 Apr 2007 10:02:33 -0700 (PDT) 4, 22 -- Received: from [192.168.159.233] (jpix-01.caltech.edu [131.215.2.133]) 4, 22 -- by earth-ox.its.caltech.edu (Postfix) with ESMTP id 79F7737047 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Fri, 27 Apr 2007 10:02:32 -0700 (PDT) 4, 22 -- Mime-Version: 1.0 (Apple Message framework v624) 4, 22 -- In-Reply-To: {200704271232.l3RCWT8d027474-at-ns.microscopy.com} 4, 22 -- References: {200704271232.l3RCWT8d027474-at-ns.microscopy.com} 4, 22 -- Content-Type: text/plain; charset=US-ASCII; format=flowed 4, 22 -- Message-Id: {844d0fa3d8c2f7e0c63c4b9fbb5479bd-at-caltech.edu} 4, 22 -- Content-Transfer-Encoding: 7bit 4, 22 -- From: Bill Tivol {tivol-at-caltech.edu} 4, 22 -- Subject: Re: [Microscopy] Terminology - please vote! 4, 22 -- Date: Fri, 27 Apr 2007 10:13:40 -0700 4, 22 -- To: microscopy-at-msa.microscopy.com 4, 22 -- X-Mailer: Apple Mail (2.624) 4, 22 -- X-Spam-Scanned: at Caltech-ITS on fire-dog by amavisd-2.4.5 ==============================End of - Headers==============================
If it is any use to people interested in "budget" scanners, I have posted an image on my webspace. It is an image of an unknown bacterium shot at 15kX on a JEOL JEM-100s on Kodak 4489 and scanned in the Microtek i900 at 1200 dpi. It was scanned at 16 bits and scaled to an 8bit range and converted to JPG. The jpg loses a bit of quality, but not much.
BTW, if anyone knows this beast, please let me know what it is. It is growing in crashing non-sterile Chlamydomonas cultures (which I raise under a fluorescent lamp in the lab for interesting test samples....)
Cheers!
Dale Callaham UMASS Amherst
} -----Original Message----- } X-from: jgross-at-neuron.uchc.edu [mailto:jgross-at-neuron.uchc.edu] } Sent: 26 April 2007 19:55 } To: keith.morris-at-ucl.ac.uk } Subject: [Microscopy] Scanners for TEM negatives } } } } } ---------------------------------------------------------------------------- } 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 } ---------------------------------------------------------------------------- } } Our lab needs to purchase a new scanner for our TEM negatives. We } have narrowed our choices to either the Epson Perfection V750 } (recommended by Keith Young in the May 2006 issue of Microscopy } Today) or the Microtek ScanMaker i900. The major difference between } the two seems to be the "glassless" scanning feature of the Microtek. } Has anyone had first-hand experience of the film scanning ability of } either one of these scanners? Or are there any quirks that would } make one of them less desirable? } } Thank you, } Julie Gross } Kent Morest Lab } Dept. of Neuroscience } UCONN Health Center } Farmington, CT 06030 } jgross-at-neuron.uchc.edu } } }
I am posting this job description for a colleague. Please respond to the Center for Nanotechnology listed below if you are interested in this position.
} Research Scientist 3 position at the University of Washigton } } } } } Position Description: } } This is a 100% position. The Research Scientist will act as a scientific } liaison and provide technical support to users of the University of } Washington Nanotech User Facility (NTUF), a node of the National } Nanotechnology Infrastructure Network (NNIN). The successful candidate } will } develop protocols on various state-of-the-art electron microscopy tools to } address demand from internal and external users and will develop new } capabilities and applications using existing equipment. This position has } potential to evolve into a managerial position upon future evaluation. } Responsibilities include: } . To act as a technical lead with a focus on transmission electron } microscopy (TEM) in support of research projects for NTUF users across the } nation. } . To enhance the capabilities of existing electron microscopy tools } and } explores new and emerging applications of these tools with the goal of } increasing NTUF usage. } . To design creative training procedures and ensure effective training } of NTUF users on the use of TEM and other NTUF tools. } . To develop and formulate sample preparation procedures in support of } users. } . To perform routine operation and maintenance on the TEM and other } NTUF } equipment } . To perform contract work for remote users nationwide } . To collaborate with and lead tasks/projects with other NTUF } personnel } in order to achieve objectives and/or enhance service quality } . To assume other duties as requested by the directors and lab } manager. } } } } } } } Requirements: } } The successful candidate is expected to have a broad knowledge of the } principle and practice of transmission electron microscopy and to } establishtechnical expertise in TEM operation, working proactively and } collaboratively with a vibrant scientific community making use of NTUF and } NNIN. } . Education: Ph.D. in Science or Engineering with 1-2 years of } professional experience or BS/MS level with 3-5 years of relevant } experience } } . Broad knowledge of nanoscience and nanotechnology tools and } applications } . Research experience in the field of TEM applications in nanoscale } science and technology, preferably in bio-related } . Experience in working with students/users in a multi-user academic } facility } . Excellent written and communication skills for preparation of } reports } and presentations } . Self motivated with a strong desire to succeed } . Excellent team player who will contribute to the team success } } } } } } --------------------------------------------------------------- } } Dong Qin, PhD MBA } } Associate Director, Center for Nanotechnology } } University of Washington, Seattle, WA 98195 } } Voice: 206-616-2118; Fax: 206-221-2528 } } {http://depts.washington.edu/ntuf} http://depts.washington.edu/ntuf } } www.nano.washington.edu } } } } } }
==============================Original Headers============================== 4, 30 -- From joswiak-at-astro.washington.edu Fri Apr 27 16:08:32 2007 4, 30 -- Received: from mimas.astro.washington.edu (mimas99.astro.washington.edu [128.95.99.14]) 4, 30 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3RL8WGN010884 4, 30 -- for {Microscopy-at-microscopy.com} ; Fri, 27 Apr 2007 16:08:32 -0500 4, 30 -- Received: from mimas.astro.washington.edu (localhost.localdomain [127.0.0.1]) 4, 30 -- by mimas.astro.washington.edu (8.12.11.20060308/8.12.11) with ESMTP id l3RL8VGm032759 4, 30 -- (version=TLSv1/SSLv3 cipher=DHE-RSA-AES256-SHA bits=256 verify=NO); 4, 30 -- Fri, 27 Apr 2007 14:08:31 -0700 4, 30 -- Received: (from apache-at-localhost) 4, 30 -- by mimas.astro.washington.edu (8.12.11.20060308/8.12.11/Submit) id l3RL8VPn032757; 4, 30 -- Fri, 27 Apr 2007 14:08:31 -0700 4, 30 -- X-Authentication-Warning: mimas.astro.washington.edu: apache set sender to joswiak-at-astro.washington.edu using -f 4, 30 -- Received: from 128.95.99.157 4, 30 -- (SquirrelMail authenticated user joswiak) 4, 30 -- by mail.astro.washington.edu with HTTP; 4, 30 -- Fri, 27 Apr 2007 14:08:31 -0700 (PDT) 4, 30 -- Message-ID: {64177.128.95.99.157.1177708111.squirrel-at-mail.astro.washington.edu} 4, 30 -- Date: Fri, 27 Apr 2007 14:08:31 -0700 (PDT) 4, 30 -- Subject: TEM specialist job 4, 30 -- From: "Dave Joswiak" {joswiak-at-astro.washington.edu} 4, 30 -- To: Microscopy-at-microscopy.com 4, 30 -- Cc: dqin-at-u.washington.edu 4, 30 -- Reply-To: "Dave" {Joswiak-at-astro.washington.edu} 4, 30 -- User-Agent: SquirrelMail/1.4.8-4.el3 4, 30 -- MIME-Version: 1.0 4, 30 -- Content-Type: text/plain;charset=iso-8859-1 4, 30 -- Content-Transfer-Encoding: 8bit 4, 30 -- X-Priority: 3 (Normal) 4, 30 -- Importance: Normal 4, 30 -- X-Greylist: Sender IP whitelisted, not delayed by milter-greylist-2.0.2 (mimas.astro.washington.edu [127.0.0.1]); Fri, 27 Apr 2007 14:08:31 -0700 (PDT) ==============================End of - Headers==============================
A veritable treatise on scanners. Thank you. I love this list! --Jan Factor
---------------------------------------------- Jan Robert Factor, Ph.D. Professor of Biology ---------------------------------------------- Natural Science Bldg. Purchase College, State University of New York 735 Anderson Hill Rd. Purchase, NY 10577 USA ---------------------------------------------- Office Tel: 914-251-6659 Office Fax: 914-251-6635 E-mail: jfactor-at-ns.purchase.edu ----------------------------------------------
keith.morris-at-ucl.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 } ---------------------------------------------------------------------------- } } Hi Julie, } } I don't know if I'm Keith Young, but to be honest Microtek haven't bought } out a new top flight scanner for years and their ArtixScan designs are } living on their old name and hardware. Agfa scanners were simply rebadged } Microtek's until they withdrew from the scanner market along with Fuji. The } Microtek ArtixScan 2500f is pretty much the £5,000 Agfa Duoscan 2550T from 7 } years ago, now with firewire instead of SCSI. Not that these professional } Microtek scanners are rubbish, just a bit expensive for a similar scan } quality as the Epson 4990 Photo (and their scans are probably a bit inferior } in resolution to the 6,400dpi Epson V750 - the Agfa 2550T and Epson 4990 } are). As I mentioned the upmarket Microtek scanners may have a far better } build quality though. The cheaper Microtek i900, introduced I think in 2004, } has a resolution of about 3,200 dpi. It's glassless design could help reduce } problems with dust, although the scanner optics is no longer 'sealed behind } glass'. It uses 4x5" film holders, a 8x10" glass sandwich or the A4 top bed } for reflective scans. It has been reported as very slow, but then most } prosumer flatbeds at this price aren't that fast. I doubt if you would be } disappointed if you bought the Microtek i900 or the Epson V750 (or the Epson } V700 or 4990 Photo for that matter). } } The only way to find out if, at 3,200 dpi quoted resolution, the i900 } produces better scans (& faster) than the 6,400 dpi quoted Epson 750 is to } scan the same negative in both, at the same and maximum resolution (I've } never tried the i900, although our Microtek/Agfa Duoscan 2550T looks very } similar). There was also the similar film only Epson F3200 [dpi], that could } scan negatives greater than 9 x 6 cm using 4x5" film holders, but it seems } to have been withdrawn recently. } } I did own an old Nikon 2,700 dpi dedicated 35mm slide scanner at home but it } simply took up too much room and it's scan quality was worse than the Canon } 9950F I got to replace it, particularly for colour negatives - plus the } Canon only does reflective scans to A4, but annoyingly Canon's Scangear } restricts film sizes to it's film holders. In comparison the V700/V750 pro } can go to A4 for film, with film in holders or just lying on the platen. I } have only used a 6,400 dpi V750 Pro briefly for 35mm colour slide film (my } main interest), and its scan quality was a bit better than my Canon 9950F, } i.e. the fuzz was a tiny tiny bit sharper, it's not streets ahead at all } (although it's Epson Scan twain interface was in another league). Plus the } 35mm slide holder retaining clips on the V750 are rather flimsy (although } the Canon and Epson 4990 don't even bother with retaining clips, using } gravity to hold them in place). I don't know how good the 35mm clips on the } i900 are (they don't look great either). At work our 4,800 dpi Epson 4990 } Photo is more than adequate for 1,200 dpi B&W TEM film scans, so I haven’t } bothered upgrading to a V750 Pro. The V750 does also offer an optional } fluid mount scan accessory like every expensive drum scanners, but that’s } unlikely to be of interest for TEM film scanning. It's meant to get rid of } Newton rings (swirls) and can assist in hiding scratches and dust, but it } often introduces bubbles instead. } } However you will get superior scans from an £8,000 Hasselblad 8,000 dpi } [Imacon] 848 semi-drum scanner, but often it will be the film grain that } looks a bit better resolved rather than actual image detail. Plus you must } use the film holders - it scans fast though. It also has no glass between } the film and detector. Things like FARE/ICE dust removal and accurate colour } reproduction (the latter being essential for professionals that need correct } skin tones or accurate reproductions of museum articles) are irrelevant for } your B&W scientific images. } } The excellent 4000 dpi 35mm Nikon Coolscan 9000 [3-CCD sensor] film scanner } comes in at £2,000. However the Nikon 9000 can only scan film up to 9 x 6 cm } in size and histological sections on glass slides (too small for most TEM } negatives unless you cut them down). Plus it has a lot of software/hardware } for 35mm colour film thrown in you probably won't need. } } To be honest the 4,800 dpi Epson 4990 Photo at £250 is more than adequate } for large format negatives (TEM film) as it can go up to A4 film (six TEM } negatives per scan) and Epson's Scan is an easier Twain interface than } Silverfast and Canons awful Scangear (there is also the highly regarded } Vuescan third party twain software available). } } In practice it is most unlikely that will often want to scan at more than } 1,200 dpi anyway as the resulting file size will be enormous and highly } prone to file corruption on the PC (but images should have noticeably better } resolution if scanned at 2,400dpi). Most scan just for publication or for PC } on-screen image analysis, rather than printing to A4/A3. A Epson 4990 Photo } 4,800 dpi scan of Kodak 4489 negative is around 250 Mb in TIF format, at } 2,400 dpi it drops to a 60Mb TIF file with little different in image } quality. There no reason however not to go for the better 6,400 dpi Epson } V750 Pro, even if you scan mainly at 1,200 dpi, as it's only a bit more } money (and not ours). The 6,400 dpi will be useful for enlargement of } selected areas (scanning at 3,200 dpi being optimal). } } Perhaps one downside to the V750 Pro is that it works best with the } slide/film holders and their little leg supports. Some sizes of TEM film may } have small parts of the image hidden by a 4x5" film holder. You can source } third party film holders though (see Doug Fisher), or you can just drop them } onto the platen and live with a possible small drop in resolution. As } mentioned you can get interference patterns (swirls) if the negative is } placed on the platen rather than using the 4x5" film holder. This is } probably caused by a slightly damp negative (high humidity) or the glass } platen having greasy finger marks. So far I have always got rid of this by } cleaning the platen (50% propanol/50% water with soft tissue and air-bulb } blow dry - also great for finger smudged VDU screens). Remove film from the } platen with a sheet of card rather than fingers (i.e. never touch the glass } platen). No-one here ever bothers to use the Epson 4990's 4x5" film holders } as it obscures part of the negative, only takes a few films and is far more } fiddly. They just place six TEM negatives directly on the platen (correct } side up). } } If you are really really serious about scanning film for some reason there } is also the likes of professional flatbed scanners such as the Kodak } ‘Creo’£15k [10,000 dpi] IQSmart 3, the £30k [8000 to 14000 dpi] EverSmart } Supreme II flatbeds (www.kodak.com). However cheaper consumer ‘photo’ flat } beds such as the new 6,400 dpi Epson V750 and V750 Pro come in at £500 and } £650 respectively, and I can't see any reason to spend more for infrequent } scans of large format B&W film, as you will rarely, if ever, scan at maximum } resolution. They also have convenient USB2 and Firewire interfaces and work } with Apples or PCs. } } You will have to spend a bit of time working on the best twain scan settings } and images will probably need some post-editing in Photoshop CS3 to get } optimal scanned image quality. } } But the choice is yours. Do however have a look at } http://www.photo-i.co.uk for Independent reviews of some prosumer large } format (A4) film scanners (not the i900). Also note that these cheaper } scanners are for light use, if you want to scan hundreds of films a day, and } not be chained to the scanner, look elsewhere and with a deeper pocket. } } Regards } } Keith } } PS. My 'Scanning film on a budget' Microscopy Today article has been updated } and reprinted in Infocus (March 2007) the UK Royal Microscopical society's } magazine. I can't send a pdf though as the editor hasn't passed on a } corrected version yet. I can only send a scan of the article taken using my } home 4,800 dpi Canon 9950F (scanned at 600 dpi) - but don't try that at home } with your £2,000 Nikon Coolscan 9000. } } PPS. As mentioned the now withdrawn Canon 9550F has a terrible twain } software interface(Scangear), the Epsons Scan one is miles better, so I } would avoid the Canon even if you can still find it [unless its only ever } being used for scanning photographs or with standard film holder sizes]. } Interestingly Canon scrapped their dedicated 35mm film scanners for the } 9950F (and they should know something about film photography) but the poor } Scangear twain interface didn't make many friends. I bought the 9950F just } before the Epson 4990 Photo came in - which I later purchased for work use, } mainly for TEM negatives, 2D gel film and reflective scans. } } } --------------------------- } Dr Keith J Morris } Imaging Facilities Manager } Cell Biology } Institute of Ophthalmology } UCL, London EC1V 9EL } } } } } } -----Original Message----- } X-from: jgross-at-neuron.uchc.edu [mailto:jgross-at-neuron.uchc.edu] } Sent: 26 April 2007 19:55 } To: keith.morris-at-ucl.ac.uk } Subject: [Microscopy] Scanners for TEM negatives } } } } } ---------------------------------------------------------------------------- } 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 } ---------------------------------------------------------------------------- } } Our lab needs to purchase a new scanner for our TEM negatives. We } have narrowed our choices to either the Epson Perfection V750 } (recommended by Keith Young in the May 2006 issue of Microscopy } Today) or the Microtek ScanMaker i900. The major difference between } the two seems to be the "glassless" scanning feature of the Microtek. } Has anyone had first-hand experience of the film scanning ability of } either one of these scanners? Or are there any quirks that would } make one of them less desirable? } } Thank you, } Julie Gross } Kent Morest Lab } Dept. of Neuroscience } UCONN Health Center } Farmington, CT 06030 } jgross-at-neuron.uchc.edu } } } } } ==============================Original Headers============================== } 32, 24 -- From keith.morris-at-ucl.ac.uk Fri Apr 27 07:11:58 2007 } 32, 24 -- Received: from smtp1.global.net.uk (smtp1.global.net.uk [80.189.94.53]) } 32, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3RCBvDs015635 } 32, 24 -- for {microscopy-at-microscopy.com} ; Fri, 27 Apr 2007 07:11:57 -0500 } 32, 24 -- Received: from 141.11.rb4.adsl.brightview.com ([80.189.11.141] helo=loungepc) } 32, 24 -- by smtp1.global.net.uk with esmtp (Exim 4.42) } 32, 24 -- id 1HhPIt-000KbJ-3b } 32, 24 -- for microscopy-at-microscopy.com; Fri, 27 Apr 2007 13:11:59 +0100 } 32, 24 -- From: "Keith Morris" {keith.morris-at-ucl.ac.uk} } 32, 24 -- To: {microscopy-at-microscopy.com} } 32, 24 -- References: {200704261855.l3QItI2m026586-at-ns.microscopy.com} } 32, 24 -- Subject: RE: [Microscopy] Scanners for TEM negatives } 32, 24 -- Date: Fri, 27 Apr 2007 13:11:57 +0100 } 32, 24 -- Message-ID: {000901c788c5$40acf4c0$0201a8c0-at-loungepc} } 32, 24 -- MIME-Version: 1.0 } 32, 24 -- Content-Type: text/plain; } 32, 24 -- charset="iso-8859-1" } 32, 24 -- X-Mailer: Microsoft Office Outlook 11 } 32, 24 -- Thread-Index: AceINHLb4TXSJai/QcGWrztdBBdnGgAHZTog } 32, 24 -- In-Reply-To: {200704261855.l3QItI2m026586-at-ns.microscopy.com} } 32, 24 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.3028 } 32, 24 -- Authenticated-Sender: } 32, 24 -- Content-Transfer-Encoding: 8bit } 32, 24 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l3RCBvDs015635 } ==============================End of - Headers============================== } } }
==============================Original Headers============================== 7, 22 -- From jfactor-at-ns.purchase.edu Fri Apr 27 19:02:34 2007 7, 22 -- Received: from mta3.srv.hcvlny.cv.net (mta3.srv.hcvlny.cv.net [167.206.4.198]) 7, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3S02WZh024559 7, 22 -- for {microscopy-at-msa.microscopy.com} ; Fri, 27 Apr 2007 19:02:34 -0500 7, 22 -- Received: from [127.0.0.1] (ool-4357e3d8.dyn.optonline.net [67.87.227.216]) 7, 22 -- by mta3.srv.hcvlny.cv.net 7, 22 -- (Sun Java System Messaging Server 6.2-6.01 (built Apr 3 2006)) 7, 22 -- with ESMTP id {0JH600JFXLG6OT60-at-mta3.srv.hcvlny.cv.net} for 7, 22 -- microscopy-at-msa.microscopy.com; Fri, 27 Apr 2007 20:02:32 -0400 (EDT) 7, 22 -- Date: Fri, 27 Apr 2007 20:02:42 -0400 7, 22 -- From: Jan Factor {jfactor-at-ns.purchase.edu} 7, 22 -- Subject: Re: [Microscopy] RE: Scanners for TEM negatives 7, 22 -- In-reply-to: {200704271212.l3RCCxb7017027-at-ns.microscopy.com} 7, 22 -- To: keith.morris-at-ucl.ac.uk, 7, 22 -- Microscopy Listserver {microscopy-at-msa.microscopy.com} 7, 22 -- Message-id: {46328F22.1010206-at-ns.purchase.edu} 7, 22 -- MIME-version: 1.0 7, 22 -- Content-type: text/plain; charset=windows-1252; format=flowed 7, 22 -- References: {200704271212.l3RCCxb7017027-at-ns.microscopy.com} 7, 22 -- User-Agent: Thunderbird 1.5.0.10 (Windows/20070221) 7, 22 -- Content-Transfer-Encoding: 8bit 7, 22 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l3S02WZh024559 ==============================End of - Headers==============================
I've been away from my desk and missed this thread. I don't like to jump in at the end but I don't agree that the Microtek is the way to go. The two problems that I have with the Microtek have been speed and resolution. Although they may have inproved the resolution some the speed issue sounds about the same. The glassless idea is ok but the only reason that you might be worried about the resolution is if you are working in the 2000 ppi range. We do do our work prints at 1200 dpi but high res scanns are at 4800. I have actually measured the resolution in transparency mode at a true 2000 pixels per inch with the 4870 and 4990 Epsons. The Epson D750 Pro is actuall better but has not been thru testing as yet. (It sits on my desk next to the 4990) The big difference is that the Epsons are 5 to 10 times faster at scanning. I was a Microtek fan but nothing has come close to the Epsons in about 5 years.
I don't have alot of time but would like to put forth a few critically important facts that we have* measured.*
1. The Silverfast driver is worthless.(The upgrade is worse than the free teaser one) It is fast and inaccurate not good features for scientific imaging. We use the slower and accurate twain driver that comes with the Epson.for free 2. Always scan negatives as positive transparencies. If you don't think this is true...Do it. The scanning range for scanning negatives is smaller than the scanning range in the positive transparency mode. 3. We have been working on holders that fully support 4 negatives. 4. We normally have a 4990 or 4870 on each computer so that scanning can be done while we do other things The D750 for --|8k x 8k 5. We insure that all "automatic" adjustments are turned off by hitting reset before every scan. 6. We insure that all filters and all histograms are truly set to zero and/or max/min This is an insidious problem. The manufacturers are turning on filters and not resetting to zero or to max in the defaults. Don't ask me why cause I don't know. They change this with every version of the drivers so you must constantly check.
If you want really high resolution then the D750 has a liquid immersion system that allows you to get higher than anything but the drum scanners.
Using the Epson we can prove that we get 8k by 8 k in 16 bit mode we see --|3.7 O.D.The negative has 16k by 16k at 12 bits The D750 with immersion might get us really close to this.(Remember the 4990 is only $450)
I will point out to those of you who may think that you don't need to calibrate your scanners that I found one of mine was getting less than half the resolution on one axis and was even worse on the other axis.An astigmatic scanner I currently have about $3,500 in test targets that I've been using to check. The very high end testing by MTF (modulation transfer function on slanted lines) requires a unique system that is currently not commercially available.
If people on the list are interested I'll try to flesh out some of this stuff soon
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
-- 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
==============================Original Headers============================== 14, 17 -- From john_mackenzie-at-ncsu.edu Fri Apr 27 23:38:16 2007 14, 17 -- Received: from ms-smtp-04.southeast.rr.com (ms-smtp-04.southeast.rr.com [24.25.9.103]) 14, 17 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3S4cGUu006801 14, 17 -- for {Microscopy-at-microscopy.com} ; Fri, 27 Apr 2007 23:38:16 -0500 14, 17 -- Received: from [127.0.0.1] (cpe-065-190-163-148.nc.res.rr.com [65.190.163.148]) 14, 17 -- by ms-smtp-04.southeast.rr.com (8.13.6/8.13.6) with ESMTP id l3S4cELh029021 14, 17 -- for {Microscopy-at-microscopy.com} ; Sat, 28 Apr 2007 00:38:15 -0400 (EDT) 14, 17 -- Message-ID: {4632CFAE.90801-at-ncsu.edu} 14, 17 -- Date: Sat, 28 Apr 2007 00:38:06 -0400 14, 17 -- From: John Mackenzie {john_mackenzie-at-ncsu.edu} 14, 17 -- User-Agent: Thunderbird 1.5.0.10 (Windows/20070221) 14, 17 -- MIME-Version: 1.0 14, 17 -- To: Microscopy-at-microscopy.com 14, 17 -- Subject: Re: Scanners for TEM Negatives 14, 17 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 14, 17 -- Content-Transfer-Encoding: 7bit 14, 17 -- X-Virus-Scanned: Symantec AntiVirus Scan Engine ==============================End of - Headers==============================
Good selections. Don't get a UMAX scanner. I have UMAX III SCSI and it has failed too many times. Factory returns garner marginal improvements.
I think that any unit other than UMAX could be a good solution. Caveat emptor.
gary g.
At 08:40 PM 4/27/2007, you wrote: } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
==============================Original Headers============================== 6, 20 -- From gary-at-gaugler.com Sat Apr 28 00:17:39 2007 6, 20 -- Received: from qsmtp1.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 l3S5HcY3018525 6, 20 -- for {microscopy-at-microscopy.com} ; Sat, 28 Apr 2007 00:17:38 -0500 6, 20 -- Message-Id: {200704280517.l3S5HcY3018525-at-ns.microscopy.com} 6, 20 -- Received: (qmail 14817 invoked from network); 27 Apr 2007 22:17:38 -0700 6, 20 -- Received: by simscan 1.1.0 ppid: 14808, pid: 14810, t: 0.1417s 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 qsmtp1 with SMTP; 27 Apr 2007 22:17:38 -0700 6, 20 -- X-Mailer: QUALCOMM Windows Eudora Version 7.1.0.9 6, 20 -- Date: Fri, 27 Apr 2007 22:17:46 -0800 6, 20 -- To: john_mackenzie-at-ncsu.edu 6, 20 -- From: Gary Gaugler {gary-at-gaugler.com} 6, 20 -- Subject: Re: [Microscopy] Re: Scanners for TEM Negatives 6, 20 -- Cc: MSA listserver {microscopy-at-microscopy.com} 6, 20 -- In-Reply-To: {200704280440.l3S4eAmN009669-at-ns.microscopy.com} 6, 20 -- References: {200704280440.l3S4eAmN009669-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-7DB860B4 ==============================End of - Headers==============================
My name should appear above the second message starting "Don't drop the humidity too low...." and not above the first message.
Ted
----- Original Message ---- X-from: Purdy Samuel Purdy {smpurdy-at-sbcglobal.net} To: drteddunne-at-yahoo.com Sent: Wednesday, April 25, 2007 4:31:31 AM
Hi John,
I haven't tried the Microtek i900, but your views echo my suspicions (that the Epson V750 Pro is probably a faster and better scanner).
I can't easily post my scans of TEM negatives using a £8,000 Hasselblad Flextight 484 and the lowly £250 Epson 4990 Photo as they come in at over 10Mb even with high jpeg compression (and my Hasselblad Flextight 484 TEM film scans CD is in the attic somewhere).
In fact for my old 35mm 100ASA Agfa colour slide film (~120 lines per mm) scan quality is virtually identical for both, with the Flextight giving miniscule extra detail and less noise with very dark shadows. The film grain is certainly resolved more clearly with the Flextight - it looks in-focus more and less fuzzy, but this doesn't actually provide much more image detail. This is probably because both scanners are resolving detail beyond that of the film. The Epson 4990 image needed some Photoshop post-editing though - ultra-sharp mask (sharpen), and shadow/highlight to resolve details in dark shadow being the main ones. If a film scan looks bad, say too dark or light, it normally means you have to adjust the scanners twain settings, which can be laborious and often needs to be done for each slide.
However I have to say that with TEM Kodak 4489 (probably better than 200 lpm) there is more detail in some areas of the image with the Flextight 484 scan. In other areas there is not a jot of difference. This could be a focussing problem with the 4990 Photo, or vibration, and/or the fact that I didn't use the 4x5" film holders, and just put the film on the glass platen (as that’s what we tend to do to allow 6 fast negative scans per go). However whether this slight extra detail is really required, given the 30 times higher price of the Hasselblad, is largely dependent to the users requirements, it would probably be lost at 1,200 dpi scanning. Plus, the Hasselblad has a yearly maintenance contract of around £800, and the cheap upmarket sibling 6,400 dpi Epson V750 Pro (£550) is even better than the 4,800 dpi 4990 Photo I use.
However given the price of a digital camera on a TEM, £8,000 for a top quality Hasselblad scanner isn't ludicrous (but you can't do A4 reflective scans so you still need that standard A4 flatbed). See your local reprographics department if you want a tour of a Flextight (but don't expect the operators to always know too much about it technically - they probably won't spend an hour or two getting the perfect scan from a crucial negative). See http://www.hasselblad.com/products/scanners.aspx for Hasselblads latest scanners (X1 - £7,500 and X5 - £12,000). The prices & some details are also at http://www.robertwhite.co.uk/hasselblad.htm#LabelHFS.
Always remember that the best scanner in the world will always degrade the image compared to original film, as going through a series of optics again will lose some ultra-fine detail - just as the original film lost detail via limitations of the camera optics/film when the image was captured. The amount of degradation is dependent on scanner optics and mechanism quality (and thus generally price, although there are no doubt expensive lemons and cheap stars out there). Thus although a digital and professional slide film SLR camera will capture similar image detail from a scene, the image from the film will always suffer when scanned into digital form. You can see this extra detail in the original film, compared to the scanned image, by simply looking at the film with an 8x magnifier over a light-box (it's not startling, but it is definitely there, and it's seen easiest with colour slide film of say writing on the side of ship, a house or our kids faces).
Rather like taking a picture with a quality compact digital camera it can look great, but compare it with the same view taken with a digital SLR and the difference in detail at high magnification can be quite startling. Your scanner may be producing good images at 1,200 dpi, but it might not match the best at maximum resolution or at that downscaled to 1,200 dpi using a quality canner.
Personally I don't actually mind Silverfast, but it's not a very easy interface. I use $120 Silverfast Ai with my Canon 9950F, as Canon's Scangear twain software is so very poor it simply makes Silverfast look good (Silverfast doesn't have Fare dust removal support though, as Canon won't tell them how it works, you only get support for Digital ICE on Epson and other scanners).
Regards
Keith
--------------------------- 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: 28 April 2007 05:43 To: keith.morris-at-ucl.ac.uk
Folks:
I've been away from my desk and missed this thread. I don't like to jump in at the end but I don't agree that the Microtek is the way to go. The two problems that I have with the Microtek have been speed and resolution. Although they may have inproved the resolution some the speed issue sounds about the same. The glassless idea is ok but the only reason that you might be worried about the resolution is if you are working in the 2000 ppi range. We do do our work prints at 1200 dpi but high res scanns are at 4800. I have actually measured the resolution in transparency mode at a true 2000 pixels per inch with the 4870 and 4990 Epsons. The Epson D750 Pro is actuall better but has not been thru testing as yet. (It sits on my desk next to the 4990) The big difference is that the Epsons are 5 to 10 times faster at scanning. I was a Microtek fan but nothing has come close to the Epsons in about 5 years.
I don't have alot of time but would like to put forth a few critically important facts that we have* measured.*
1. The Silverfast driver is worthless.(The upgrade is worse than the free teaser one) It is fast and inaccurate not good features for scientific imaging. We use the slower and accurate twain driver that comes with the Epson.for free 2. Always scan negatives as positive transparencies. If you don't think this is true...Do it. The scanning range for scanning negatives is smaller than the scanning range in the positive transparency mode. 3. We have been working on holders that fully support 4 negatives. 4. We normally have a 4990 or 4870 on each computer so that scanning can be done while we do other things The D750 for --|8k x 8k 5. We insure that all "automatic" adjustments are turned off by hitting reset before every scan. 6. We insure that all filters and all histograms are truly set to zero and/or max/min This is an insidious problem. The manufacturers are turning on filters and not resetting to zero or to max in the defaults. Don't ask me why cause I don't know. They change this with every version of the drivers so you must constantly check.
If you want really high resolution then the D750 has a liquid immersion system that allows you to get higher than anything but the drum scanners.
Using the Epson we can prove that we get 8k by 8 k in 16 bit mode we see --|3.7 O.D.The negative has 16k by 16k at 12 bits The D750 with immersion might get us really close to this.(Remember the 4990 is only $450)
I will point out to those of you who may think that you don't need to calibrate your scanners that I found one of mine was getting less than half the resolution on one axis and was even worse on the other axis.An astigmatic scanner I currently have about $3,500 in test targets that I've been using to check. The very high end testing by MTF (modulation transfer function on slanted lines) requires a unique system that is currently not commercially available.
If people on the list are interested I'll try to flesh out some of this stuff soon
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
-- 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
==============================Original Headers============================== 14, 17 -- From john_mackenzie-at-ncsu.edu Fri Apr 27 23:38:16 2007 14, 17 -- Received: from ms-smtp-04.southeast.rr.com (ms-smtp-04.southeast.rr.com [24.25.9.103]) 14, 17 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3S4cGUu006801 14, 17 -- for {Microscopy-at-microscopy.com} ; Fri, 27 Apr 2007 23:38:16 -0500 14, 17 -- Received: from [127.0.0.1] (cpe-065-190-163-148.nc.res.rr.com [65.190.163.148]) 14, 17 -- by ms-smtp-04.southeast.rr.com (8.13.6/8.13.6) with ESMTP id l3S4cELh029021 14, 17 -- for {Microscopy-at-microscopy.com} ; Sat, 28 Apr 2007 00:38:15 -0400 (EDT) 14, 17 -- Message-ID: {4632CFAE.90801-at-ncsu.edu} 14, 17 -- Date: Sat, 28 Apr 2007 00:38:06 -0400 14, 17 -- From: John Mackenzie {john_mackenzie-at-ncsu.edu} 14, 17 -- User-Agent: Thunderbird 1.5.0.10 (Windows/20070221) 14, 17 -- MIME-Version: 1.0 14, 17 -- To: Microscopy-at-microscopy.com 14, 17 -- Subject: Re: Scanners for TEM Negatives 14, 17 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 14, 17 -- Content-Transfer-Encoding: 7bit 14, 17 -- X-Virus-Scanned: Symantec AntiVirus Scan Engine ==============================End of - Headers==============================
==============================Original Headers============================== 38, 24 -- From keith.morris-at-ucl.ac.uk Sat Apr 28 06:42:39 2007 38, 24 -- Received: from smtp1.global.net.uk (smtp1.global.net.uk [80.189.94.53]) 38, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3SBgcNP014469 38, 24 -- for {Microscopy-at-microscopy.com} ; Sat, 28 Apr 2007 06:42:39 -0500 38, 24 -- Received: from 162.123.gr5.adsl.brightview.com ([80.189.123.162] helo=loungepc) 38, 24 -- by smtp1.global.net.uk with esmtp (Exim 4.42) 38, 24 -- id 1HhlK4-0006J2-Ka 38, 24 -- for Microscopy-at-microscopy.com; Sat, 28 Apr 2007 12:42:40 +0100 38, 24 -- From: "Keith Morris" {keith.morris-at-ucl.ac.uk} 38, 24 -- To: {Microscopy-at-microscopy.com} 38, 24 -- References: {200704280442.l3S4gbQV013302-at-ns.microscopy.com} 38, 24 -- Subject: RE: [Microscopy] Re: Scanners for TEM Negatives 38, 24 -- Date: Sat, 28 Apr 2007 12:42:39 +0100 38, 24 -- Message-ID: {000c01c7898a$5341ecb0$0301a8c0-at-loungepc} 38, 24 -- MIME-Version: 1.0 38, 24 -- Content-Type: text/plain; 38, 24 -- charset="iso-8859-1" 38, 24 -- X-Mailer: Microsoft Office Outlook 11 38, 24 -- In-reply-to: {200704280442.l3S4gbQV013302-at-ns.microscopy.com} 38, 24 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.3028 38, 24 -- Thread-Index: AceJT6ePEBeTQ9hcTRKANwzrfkHvZgAL0pgg 38, 24 -- Authenticated-Sender: 38, 24 -- Content-Transfer-Encoding: 8bit 38, 24 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l3SBgcNP014469 ==============================End of - Headers==============================
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Email: {PRICE-at-gw.med.sc.edu} Name: "Bob Price"
Organization: University of South Carolina
Title-Subject: [Filtered] Confocal Workshop
Question: The University of South Carolina Instrumentation Resource Facility and the South Carolina EPSCoR (NSF) and INBRE (NIH) programs will be hosting a hands on workshop in Basic Confocal Microscopy from June 18-21. Full details concerning the workshop and registration information can be found at http://www.scepscor.org/outreach/workshops/confocal-microscopy/home.html
The workshop will consist of both lectures and laboratory sessions and is directed towards beginning and intermediate level users of confocal microscopy. Topics covered will include the basics of fluorescence and selection of probes, specimen fixation and staining, proper set up of operating parameters for collection of images, composition of digital confocal images, the use of image enhancement and analysis programs such as Photoshop and Metamorph for confocal images, and the use of 3-D reconstruction programs such as AMIRA.
Instructors for the workshop will include Dr. Jay Jerome from Vanderbilt University, Dr. John Mackenzie from North Carolina State University, Dr. Tom Trusk from the Medical University of South Carolina, and Drs. Bob Price and John Fuseler from the USC School of Medicine.
A variety of confocal microscopes including the BD CARV spinning disk, Leica, Nikon, Olympus, and Zeiss systems are scheduled to be on hand for the laboratory sessions. Applications experts from each of the companies will also be on hand for discussion of their specific systems. Participants are encouraged to bring their own specimens for sample preparation and imaging sessions, but a variety of cell culture and tissue sections will also be available for laboratory sessions on fluorescent staining and imaging.
For further information or questions please contact Bob Price (Price-at-med.sc.edu)
Workshop on Basic Confocal Microscopy Monday June 18, 2007 8:30-9:00 Registration and Continental Breakfast, Bldg 1, Room B59 USC School of Medicine 9:00-9:15 Welcome and Logistics 9:15-10:15 Introduction and Overview of Confocal Microscopy: Jay Jerome (Vanderbilt University) and Bob Price (USC School of Medicine) 10:15-10:45 Break and Discussion 10:45-12:00 Basics of Fluorescence, Dye Characteristics: Jerome and Price 12:00-1:00 Lunch (Provided) and discussion 1:00-3:00 Specimen Preparation: Jerome and Price 3:00-3:30 Break and Discussion 3:30-?? Lab * Specimen Preparation and processing: Processing of own samples or samples will be provided. Fixation through overnight primary antibody incubation Dinner on your own
Tuesday June 19, 2007 8:30-9:00 Continental Breakfast * Room B59 9:00-10:00 Lab - Wash specimens and secondary antibody incubation 10:00-10:30 Break and Discussion 10:30-12:00 Components, proper set up of operating parameters, and types of confocal microscopes: Jerome and Price 12:00-1:30 Lunch (Provided) and Specimen Processing 1:30-2:30 Digital Images in Confocal Microscopy * Jerome and Price 2:30-4:00 Lab * finish specimen processing 4:00-6:00 Time on Instruments (BD CARV, Leica, Nikon, Olympus, Zeiss systems are scheduled to be available) Dinner on your own
Wednesday June 20, 2007 8:30-9:00 Continental Breakfast * Room B59 9:00-12:00 Photoshop, etc with confocal images (John Mackenzie * North Carolina State University) 12:00-1:00 Lunch 1:00-5:00 Time on Confocal Instruments and Software 6:00-?? Reception on Lake Murray
Thursday June 21, 2007 8:30-9:00 Continental Breakfast * Room B59 9:00-11:00 3-D reconstruction of confocal data sets with AMIRA (Tom Trusk * Medical University of South Carolina) 11:00-12:00 Ethics in Use of Confocal Images, Available Resources, Discussion 12:00 - 4:00 Lunch and additional time on Confocal Instruments and Software (Photoshop, AMIRA, Metamorph)
Robert L. Price, PhD Research Professor and Director, Instrumentation Resource Facility School of Medicine, USC Bldg 1 Room B60 6439 Garner's Ferry Road Columbia, SC 29208
It is in fact quite important to insure that the negatives are held at a uniform 1mm distance. This is why we went to trouble to machine special holders. It is odd to make a $150 holder for a $450 scanner but 3.25 by 4.00 is an odd size. It was particularly upsetting as the first prototypes didn't work and would only scan the center area. You can buy extra 4 x 5 holders for $5.00 but you have to watch that the free edge is not bent.
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
Gary Gaugler wrote: } Good selections. Don't get a UMAX scanner. I have } UMAX III SCSI and it has failed too many times. } Factory returns garner marginal improvements. } } I think that any unit other than UMAX could be } a good solution. Caveat emptor. } } gary g. } } } At 08:40 PM 4/27/2007, 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 } } ---------------------------------------------------------------------------- } } } } } } Folks: } } } } I've been away from my desk and missed this thread. I don't like to jump } } in at the end but I don't agree that the Microtek is the way to go. The } } two problems that I have with the Microtek have been speed and } } resolution. Although they may have inproved the resolution some the } } speed issue sounds about the same. The glassless idea is ok but the only } } reason that you might be worried about the resolution is if you are } } working in the 2000 ppi range. We do do our work prints at 1200 dpi but } } high res scanns are at 4800. I have actually measured the resolution in } } transparency mode at a true 2000 pixels per inch with the 4870 and 4990 } } Epsons. The Epson D750 Pro is actuall better but has not been thru } } testing as yet. (It sits on my desk next to the 4990) The big difference } } is that the Epsons are 5 to 10 times faster at scanning. I was a } } Microtek fan but nothing has come close to the Epsons in about 5 years. } } } } I don't have alot of time but would like to put forth a few critically } } important facts that we have* measured.* } } } } 1. The Silverfast driver is worthless.(The upgrade is worse than the } } free teaser one) It is fast and inaccurate not good features for } } scientific imaging. We use the slower and accurate twain driver that } } comes with the Epson.for free } } 2. Always scan negatives as positive transparencies. If you don't think } } this is true...Do it. The scanning range for scanning negatives is } } smaller than the scanning range in the positive transparency mode. } } 3. We have been working on holders that fully support 4 negatives. } } 4. We normally have a 4990 or 4870 on each computer so that scanning can } } be done while we do other things The D750 for --|8k x 8k } } 5. We insure that all "automatic" adjustments are turned off by hitting } } reset before every scan. } } 6. We insure that all filters and all histograms are truly set to zero } } and/or max/min This is an insidious problem. The manufacturers are } } turning on filters and not resetting to zero or to max in the defaults. } } Don't ask me why cause I don't know. They change this with every version } } of the drivers so you must constantly check. } } } } If you want really high resolution then the D750 has a liquid immersion } } system that allows you to get higher than anything but the drum } } scanners. } } } } Using the Epson we can prove that we get 8k by 8 k in 16 bit mode we see } } --|3.7 O.D.The negative has 16k by 16k at 12 bits The D750 with } } immersion } } might get us really close to this.(Remember the 4990 is only $450) } } } } I will point out to those of you who may think that you don't need to } } calibrate your scanners that I found one of mine was getting less than } } half the resolution on one axis and was even worse on the other axis.An } } astigmatic scanner I currently have about $3,500 in test targets that } } I've been using to check. The very high end testing by MTF (modulation } } transfer function on slanted lines) requires a unique system that is } } currently not commercially available. } } } } If people on the list are interested I'll try to flesh out some of this } } stuff soon } } } } 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 } } } } -- } } 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 } } } } } } } } ==============================Original } } Headers============================== } } 14, 17 -- From john_mackenzie-at-ncsu.edu Fri Apr 27 23:38:16 2007 } } 14, 17 -- Received: from ms-smtp-04.southeast.rr.com } } (ms-smtp-04.southeast.rr.com [24.25.9.103]) } } 14, 17 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with } } ESMTP id l3S4cGUu006801 } } 14, 17 -- for {Microscopy-at-microscopy.com} ; Fri, 27 Apr 2007 } } 23:38:16 -0500 } } 14, 17 -- Received: from [127.0.0.1] } } (cpe-065-190-163-148.nc.res.rr.com [65.190.163.148]) } } 14, 17 -- by ms-smtp-04.southeast.rr.com (8.13.6/8.13.6) with } } ESMTP id l3S4cELh029021 } } 14, 17 -- for {Microscopy-at-microscopy.com} ; Sat, 28 Apr 2007 } } 00:38:15 -0400 (EDT) } } 14, 17 -- Message-ID: {4632CFAE.90801-at-ncsu.edu} } } 14, 17 -- Date: Sat, 28 Apr 2007 00:38:06 -0400 } } 14, 17 -- From: John Mackenzie {john_mackenzie-at-ncsu.edu} } } 14, 17 -- User-Agent: Thunderbird 1.5.0.10 (Windows/20070221) } } 14, 17 -- MIME-Version: 1.0 } } 14, 17 -- To: Microscopy-at-microscopy.com } } 14, 17 -- Subject: Re: Scanners for TEM Negatives } } 14, 17 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed } } 14, 17 -- Content-Transfer-Encoding: 7bit } } 14, 17 -- X-Virus-Scanned: Symantec AntiVirus Scan Engine } } ==============================End of - } } Headers============================== }
==============================Original Headers============================== 8, 20 -- From john_mackenzie-at-ncsu.edu Sat Apr 28 12:32:16 2007 8, 20 -- Received: from ms-smtp-03.southeast.rr.com (ms-smtp-03.southeast.rr.com [24.25.9.102]) 8, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3SHWG6i012750 8, 20 -- for {Microscopy-at-microscopy.com} ; Sat, 28 Apr 2007 12:32:16 -0500 8, 20 -- Received: from [127.0.0.1] (cpe-065-190-163-148.nc.res.rr.com [65.190.163.148]) 8, 20 -- by ms-smtp-03.southeast.rr.com (8.13.6/8.13.6) with ESMTP id l3SHVw0l014291; 8, 20 -- Sat, 28 Apr 2007 13:31:59 -0400 (EDT) 8, 20 -- Message-ID: {46338505.4010603-at-ncsu.edu} 8, 20 -- Date: Sat, 28 Apr 2007 13:31:49 -0400 8, 20 -- From: John Mackenzie {john_mackenzie-at-ncsu.edu} 8, 20 -- User-Agent: Thunderbird 1.5.0.10 (Windows/20070221) 8, 20 -- MIME-Version: 1.0 8, 20 -- To: Gary Gaugler {gary-at-gaugler.com} 8, 20 -- CC: Microscopy-at-microscopy.com 8, 20 -- Subject: Re: [Microscopy] Re: Scanners for TEM Negatives 8, 20 -- References: {200704280440.l3S4eAmN009669-at-ns.microscopy.com} {200704280517.l3S5HcUu016547-at-server-autogenerated.uni06mr.unity.ncsu.edu} 8, 20 -- In-Reply-To: {200704280517.l3S5HcUu016547-at-server-autogenerated.uni06mr.unity.ncsu.edu} 8, 20 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 8, 20 -- Content-Transfer-Encoding: 7bit 8, 20 -- X-Virus-Scanned: Symantec AntiVirus Scan Engine ==============================End of - Headers==============================
Dear all, I am wonder anyone have been using the SEM-EDS to study the characteristic of suspended particulate(0.45um filter paper retained suspended particulate) in water column relate to suspended particulate trace metals or any reference on this topic, cause mostly i get from journal is airborne particle. i already have a set of data on EDS of my sample and SEM image on my sample as well. I am having a hard time to find the journal report the suspended particulate in water column. It will be great to heard respond from u all. Thanks for your valuable time.
Have a nice day.
regard William Wong
==============================Original Headers============================== 4, 26 -- From williamchai1979-at-gmail.com Sat Apr 28 20:39:26 2007 4, 26 -- Received: from qb-out-0506.google.com (qb-out-0506.google.com [72.14.204.239]) 4, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3T1dQKB001046 4, 26 -- for {Microscopy-at-microscopy.com} ; Sat, 28 Apr 2007 20:39:26 -0500 4, 26 -- Received: by qb-out-0506.google.com with SMTP id q12so4912638qbq 4, 26 -- for {Microscopy-at-microscopy.com} ; Sat, 28 Apr 2007 18:39:26 -0700 (PDT) 4, 26 -- DKIM-Signature: a=rsa-sha1; c=relaxed/relaxed; 4, 26 -- d=gmail.com; s=beta; 4, 26 -- h=domainkey-signature:received:received:message-id:date:from:to:subject:mime-version:content-type:content-transfer-encoding:content-disposition; 4, 26 -- b=B4wMPWm8POfO4HV7l8svhE5wLRBErgz+EyirjyxunMPTuJ5mf07U+HQfBnFhLF8t4zghz6w4tstzaWnH0MtW6dT5wQ/b7QuXutThPSuiJ/N8ZR9x6RNNwRnrbb0oHqXna56KTI1zjhzmQBZ63cXMf3sliblm8h+A9YNgxyoAgvU= 4, 26 -- DomainKey-Signature: a=rsa-sha1; c=nofws; 4, 26 -- d=gmail.com; s=beta; 4, 26 -- h=received:message-id:date:from:to:subject:mime-version:content-type:content-transfer-encoding:content-disposition; 4, 26 -- b=aud4oucRHdJmRxlyffL3Xps1ocuQejEQAFs9dAfsnqSoaE+Wz08NX3MEG58LLeMng/f6v+hGhVBxSHu3AAtlQMBA27OJM/rWtkRnM/cbBOVTo0uBDAg1tfst1dKgwS+hbk9GL6eo8oT5v29sbjxiOUjaN9ZTDShAa6gHAKlUSr0= 4, 26 -- Received: by 10.70.39.11 with SMTP id m11mr8174421wxm.1177810766243; 4, 26 -- Sat, 28 Apr 2007 18:39:26 -0700 (PDT) 4, 26 -- Received: by 10.70.53.6 with HTTP; Sat, 28 Apr 2007 18:39:26 -0700 (PDT) 4, 26 -- Message-ID: {57a6a78b0704281839n370e3e1bif4f76390f72b5595-at-mail.gmail.com} 4, 26 -- Date: Sun, 29 Apr 2007 09:39:26 +0800 4, 26 -- From: "william wong" {williamchai1979-at-gmail.com} 4, 26 -- To: Microscopy-at-microscopy.com 4, 26 -- Subject: SEM-EDS for suspended particulate in water column 4, 26 -- MIME-Version: 1.0 4, 26 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 4, 26 -- Content-Transfer-Encoding: 7bit 4, 26 -- Content-Disposition: inline ==============================End of - Headers==============================
I agree it is important to use the holders for important scans at high resolution (greater than 800 dpi), although these modern flatbed film scanners do have a pretty good depth of field for low resolution scans - you can happily scan toys, leaves, seeds etc..).
Everyone here shuns the film holders as they tend to do 800 dpi scans only (rarely even 1200 dpi). I mention the holders to all our users but they always decline on the grounds that the images look fine for their purpose (thus they basically can't be bothered - even after spending weeks getting the samples to the TEM negative stage). However invariably in such cases they generally only require a very small photo for a publication or talk, the magnification being done quite rightly at the TEM rather than when scanning the film. I only invariably use the film holders when scanning TEM negatives above 800 dpi (and that's only been for the odd scanning film article I have written) - my interest is almost exclusively the scanning of 35mm colour slide film, that come in their own little holders, or 35mm colour negatives that have to be fitted into the 35mm film strip holder unless you want a really bad time in Photoshop.
Those interested in things like counting gold particles or measuring distances with TEM images via MetaMorph can occasionally be coerced into using the 4x5" holders. As I run the confocal/time-lapse/laser dissection light microscopy side of things at the institute, the scanner we use is provided for the TEM users merely as an occasional service for old film. Apple fans here still stick to our old Agfa [Microtek] 2550T scanner with it's glass sandwich system, as it's attached to an cuddly Apple (plus they paid the £5,000 for it back in 2000).
Their TEM microscope has now been converted to a rather expensive Gatan digital camera system and wet EM film processing has largely ceased. I have to say that the TEM film doesn't flatten completely in the Epson 4x5" holders, the film being only held on three sides. Plus they can only hold two smaller Kodak negatives at a time and obscure bits of the image. I would think that our workshop would easily be able to make a better version to hold more negatives fairly easily, but the requirement for scanning TEM film is now on the wane here. My main concern would be the long term archiving and storage of these very large digital images, as modern TEM film was such a stable medium in comparison, but that isn't really my problem.
Regards
Keith
-----Original Message----- X-from: john_mackenzie-at-ncsu.edu [mailto:john_mackenzie-at-ncsu.edu] Sent: 28 April 2007 18:40 To: keith.morris-at-ucl.ac.uk
Gary:
It is in fact quite important to insure that the negatives are held at a uniform 1mm distance. This is why we went to trouble to machine special holders. It is odd to make a $150 holder for a $450 scanner but 3.25 by 4.00 is an odd size. It was particularly upsetting as the first prototypes didn't work and would only scan the center area. You can buy extra 4 x 5 holders for $5.00 but you have to watch that the free edge is not bent.
John
John M. Mackenzie, Jr., PhD Professor of Microbiology Coordinator, Center for Electron Microscopy North Carolina State University Phone (919) 515-2664 Fax (919) 515-8293
==============================Original Headers============================== 18, 22 -- From keith.morris-at-ucl.ac.uk Sun Apr 29 08:53:41 2007 18, 22 -- Received: from smtp2.global.net.uk (smtp2.global.net.uk [80.189.94.52]) 18, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3TDrePF005386 18, 22 -- for {Microscopy-at-microscopy.com} ; Sun, 29 Apr 2007 08:53:41 -0500 18, 22 -- Received: from 135.186.adsl.brightview.com ([80.189.186.135] helo=loungepc) 18, 22 -- by smtp2.global.net.uk with esmtp (Exim 4.42) 18, 22 -- id 1Hi9qN-0005ze-Q1 18, 22 -- for Microscopy-at-microscopy.com; Sun, 29 Apr 2007 14:53:39 +0100 18, 22 -- From: "Keith Morris" {keith.morris-at-ucl.ac.uk} 18, 22 -- To: {Microscopy-at-microscopy.com} 18, 22 -- Subject: FW: [Microscopy] Scanners for TEM Negatives 18, 22 -- Date: Sun, 29 Apr 2007 14:53:43 +0100 18, 22 -- Message-ID: {000401c78a65$ccfc1130$0201a8c0-at-loungepc} 18, 22 -- MIME-Version: 1.0 18, 22 -- Content-Type: text/plain; 18, 22 -- charset="iso-8859-1" 18, 22 -- X-Mailer: Microsoft Office Outlook 11 18, 22 -- X-Mimeole: Produced By Microsoft MimeOLE V6.00.2900.3028 18, 22 -- Thread-Index: AceJvFNv1dhyj4vRT0CRALhngBBB/QAgku0gAAnF/iA= 18, 22 -- Authenticated-Sender: 18, 22 -- Content-Transfer-Encoding: 8bit 18, 22 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l3TDrePF005386 ==============================End of - Headers==============================
We have just acquired a Hitachi S4300 SE/N and I was wondering if anyone had captured the video signal from one of these SEM's. If so, what method was used and found to be effective - capture to tape and then conversion to digital or direct capture to PC via a frame grabber.
Any ideas, pit-falls, card types etc would be much appreciated. I believe the output is NTSC.
Cheers and thank you.
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
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==============================Original Headers============================== 9, 28 -- From Colin.Veitch-at-csiro.au Mon Apr 30 00:29:33 2007 9, 28 -- Received: from act-MTAout6.csiro.au (act-MTAout6.csiro.au [150.229.7.43]) 9, 28 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3U5TVdE032721 9, 28 -- for {microscopy-at-msa.microscopy.com} ; Mon, 30 Apr 2007 00:29:32 -0500 9, 28 -- DomainKey-Signature: s=email; d=csiro.au; c=nofws; q=dns; b=idOoQ93qExsE0Aee2cmw3k5NnJKeizlphV/QVmRlbKRFbAZQKdJmHUFP9vqWKGKFX6MrpUiCkoCxTVCIoKKbbFOaIuic9LzS6AG6z2C57Fk+CTtMBybZDEmRlogpu/09; 9, 28 -- Received: from exgw1-mel.nexus.csiro.au ([138.194.3.56]) 9, 28 -- by act-ironport-int.csiro.au with ESMTP; 30 Apr 2007 15:29:33 +1000 9, 28 -- X-IronPort-AV: i="4.14,467,1170594000"; 9, 28 -- d="scan'208"; a="155855520:sNHT169288252" 9, 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); 9, 28 -- Mon, 30 Apr 2007 15:29:29 +1000 9, 28 -- X-MimeOLE: Produced By Microsoft Exchange V6.0.6603.0 9, 28 -- content-class: urn:content-classes:message 9, 28 -- MIME-Version: 1.0 9, 28 -- Content-Type: text/plain; 9, 28 -- charset="us-ascii" 9, 28 -- Subject: Video out from a Hitachi S4300SE/N 9, 28 -- Date: Mon, 30 Apr 2007 15:29:29 +1000 9, 28 -- Message-ID: {32CDDDAA7161394599F002549491574922B85D-at-exvic5-gex.nexus.csiro.au} 9, 28 -- X-MS-Has-Attach: 9, 28 -- X-MS-TNEF-Correlator: 9, 28 -- Thread-Topic: Video out from a Hitachi S4300SE/N 9, 28 -- Thread-Index: AceK6IZmauEAJ24hSzOoNHfnLkQ0Tw== 9, 28 -- From: {Colin.Veitch-at-csiro.au} 9, 28 -- To: {microscopy-at-msa.microscopy.com} 9, 28 -- X-OriginalArrivalTime: 30 Apr 2007 05:29:29.0599 (UTC) FILETIME=[8671A8F0:01C78AE8] 9, 28 -- Content-Transfer-Encoding: 8bit 9, 28 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l3U5TVdE032721 ==============================End of - Headers==============================
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Email: charlesping-at-hotmail.com Name: Charles
Organization: University of York
Title-Subject: [Filtered] The pins definition for Varian gun unit
Question: Dear all
I am going to use the Varian electron gun control unit 981-2745 and scanning sample positioner 981-2757A to control the electron gun we built. In the manual we didn't find any information for the pin definition for the cable connecting the gun control unit with the electron gun , either the gun scanning unit with the electron gun. Did anyone use this two units before? Could you tell me the pins definition of these two units' connectors to the gun? Many thanks
Charles Department of Electronics University of York UK
Drinking water Waste water Surface runoff Inland Surface water bodies Ocean water bodies
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: williamchai1979-at-gmail.com [mailto:williamchai1979-at-gmail.com] Sent: Saturday, April 28, 2007 9:46 PM To: ph2-at-sprynet.com
Dear all, I am wonder anyone have been using the SEM-EDS to study the characteristic of suspended particulate(0.45um filter paper retained suspended particulate) in water column relate to suspended particulate trace metals or any reference on this topic, cause mostly i get from journal is airborne particle. i already have a set of data on EDS of my sample and SEM image on my sample as well. I am having a hard time to find the journal report the suspended particulate in water column. It will be great to heard respond from u all. Thanks for your valuable time.
Have a nice day.
regard William Wong
==============================Original Headers============================== 4, 26 -- From williamchai1979-at-gmail.com Sat Apr 28 20:39:26 2007 4, 26 -- Received: from qb-out-0506.google.com (qb-out-0506.google.com [72.14.204.239]) 4, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3T1dQKB001046 4, 26 -- for {Microscopy-at-microscopy.com} ; Sat, 28 Apr 2007 20:39:26 -0500 4, 26 -- Received: by qb-out-0506.google.com with SMTP id q12so4912638qbq 4, 26 -- for {Microscopy-at-microscopy.com} ; Sat, 28 Apr 2007 18:39:26 -0700 (PDT) 4, 26 -- DKIM-Signature: a=rsa-sha1; c=relaxed/relaxed; 4, 26 -- d=gmail.com; s=beta; 4, 26 -- h=domainkey-signature:received:received:message-id:date:from:to:subject:mime -version:content-type:content-transfer-encoding:content-disposition; 4, 26 -- b=B4wMPWm8POfO4HV7l8svhE5wLRBErgz+EyirjyxunMPTuJ5mf07U+HQfBnFhLF8t4zghz6w4ts tzaWnH0MtW6dT5wQ/b7QuXutThPSuiJ/N8ZR9x6RNNwRnrbb0oHqXna56KTI1zjhzmQBZ63cXMf3 sliblm8h+A9YNgxyoAgvU= 4, 26 -- DomainKey-Signature: a=rsa-sha1; c=nofws; 4, 26 -- d=gmail.com; s=beta; 4, 26 -- h=received:message-id:date:from:to:subject:mime-version:content-type:content -transfer-encoding:content-disposition; 4, 26 -- b=aud4oucRHdJmRxlyffL3Xps1ocuQejEQAFs9dAfsnqSoaE+Wz08NX3MEG58LLeMng/f6v+hGhV BxSHu3AAtlQMBA27OJM/rWtkRnM/cbBOVTo0uBDAg1tfst1dKgwS+hbk9GL6eo8oT5v29sbjxiOU jaN9ZTDShAa6gHAKlUSr0= 4, 26 -- Received: by 10.70.39.11 with SMTP id m11mr8174421wxm.1177810766243; 4, 26 -- Sat, 28 Apr 2007 18:39:26 -0700 (PDT) 4, 26 -- Received: by 10.70.53.6 with HTTP; Sat, 28 Apr 2007 18:39:26 -0700 (PDT) 4, 26 -- Message-ID: {57a6a78b0704281839n370e3e1bif4f76390f72b5595-at-mail.gmail.com} 4, 26 -- Date: Sun, 29 Apr 2007 09:39:26 +0800 4, 26 -- From: "william wong" {williamchai1979-at-gmail.com} 4, 26 -- To: Microscopy-at-microscopy.com 4, 26 -- Subject: SEM-EDS for suspended particulate in water column 4, 26 -- MIME-Version: 1.0 4, 26 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 4, 26 -- Content-Transfer-Encoding: 7bit 4, 26 -- Content-Disposition: inline ==============================End of - Headers==============================
I know that this topic arises at lease once a year, but software advances so quickly nowadays that one- or two-year-old information is often completely useless.
I'm seeking opinions and information on current methods for largely automated particle classification based on color and shape/size in images. I don't need an image acquisition system, and a gallery or database feature just would be a bonus.
As I mentioned, I fear most of my information is outdated and comes mostly from advertisements or product brochures. Before we were just exploring what was out there, but now we have a real project for using such software.
I know of a few options, like Soft Imaging System's analySIS software, but its steep price tag to get all those bells and whistles is likely going to be too high for the project.
I'd be interesting in hearing from colleagues about their experiences (good and bad) with any programs in particular, and any sales people can feel free to contact me with information as well.
Thanks all in advance,
Ellery
-------------------- Ellery E. Frahm Research Fellow & Manager Electron Microprobe Laboratory University of Minnesota - Twin Cities Department of Geology & Geophysics Lab Website: http://probelab.geo.umn.edu Personal Website: http://umn.edu/~frah0010
} The question of providing cooling water for an oil diffusion pump is } discussed in some detail in Sec. 5.8.1 (p. 215) of my book 'Vacuum } Methods in Electron Microscopy' (available from SPI Supplies, M. E. } Taylor, Ladd, etc). Basically, the primary requirement is to keep } the temperature of the body of the diffusion pump at a temperature } low enough to condense the vapor of the diffusion pump fluid as it } strikes the wall of the pump . The pressure and } flow rates are only secondary matters of concern as long as this } requirement is met. I suggest you check the temperature of the } cooling coils at the top of the pump, and of the water coming out of } the water line. The water should be entering the pump at the top of } the cooling coils and exiting from the bottom, so that the top of } the pump wall is coolest. If the temperature there is "cool to the } touch" it is probably all right, because most modern diffusion pump } oils have rather low vapor pressures and readily condense at } moderately cool temperatures - liquid nitrogen is not needed. The } water flowing out of the bottom of the pump may be slightly warm to } the touch, and the pump can still function properly. } } I do not recommend using water-glycol mixtures, for reasons given in } my book. Instead I think it is better to use just distilled water, } adding a bit of algaecide to prevent algae from growing in the system. We used the algaecide Chloramine-T (the sodium salt of N-chloro-p-toluenesulphonamide, available from specialty chemical companies such as Alfa-Aldrich, Sigma, etc) at the level of 1 gram per gallon for years with very satisfactory results. Algaecides are also available from water bed and swimming pool suppliers. Using } tubes and hoses in the circulating system that are } completely opaque to light, and covering the reservoir to exclude light, will also effectively limit growth of algae in the system. You should, of course, have a good filter on the inlet to the supply line to catch any } algae (or other debris) that does happen to get into the system. -- -- 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
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==============================
All, For what it's worth, here's my two cents. I have found, after many years of experimenting with all sorts of solutions and chemicals, that the best coolant is pure distilled water.
But what about algae one might ask. Here's the secret: simply make sure to replace all lines with black (opaque) rubber hoses and that all sight glasses, flow meters and other light "windows" have a removable cover of some sorts.
If you remove all sources of light, that green scummy algae will never grow in your system! On the other hand when even a little light is present, I've had algae grow in pure ethylene glycol! The stuff mutates effortlessly it seems.
I've run half a dozen instruments this way for over ten years and never seen any algae buildup (maybe just a few tiny rust flakes here and there at the bottom of the chiller reservoir).
John J. Donovan donovan-at-uoregon.edu University of Oregon (541) 346-4632 (office) 1260 Franklin Blvd (541) 346-4655 (probe) Eugene, OR (541) 346-4778 (SEM) 97403-1272 (541) 346-4692 (FAX)
Of all the various methods and opinions that arise relative to this subject, I have to agree with you. Use distilled water, change it frequently, clean the recirculating filter, for extended chiller life, and the instruments won't be routinely stress-tested with overheating.
Bob Roberts EM Lab Services, Inc. 449 NW 62nd St. Topeka, Kansas 66617-1780 785.246.1232 voice 785.246.0168 fax www.emlabservices.com
==============================Original Headers============================== 5, 27 -- From emlabservices-at-cox.net Mon Apr 30 18:04:29 2007 5, 27 -- Received: from eastrmmtao106.cox.net (eastrmmtao106.cox.net [68.230.240.48]) 5, 27 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3UN4T8K002795 5, 27 -- for {microscopy-at-microscopy.com} ; Mon, 30 Apr 2007 18:04:29 -0500 5, 27 -- Received: from eastrmimpo02.cox.net ([68.1.16.120]) 5, 27 -- by eastrmmtao106.cox.net 5, 27 -- (InterMail vM.7.05.02.00 201-2174-114-20060621) with ESMTP 5, 27 -- id {20070430230429.EJCW12193.eastrmmtao106.cox.net-at-eastrmimpo02.cox.net} 5, 27 -- for {microscopy-at-microscopy.com} ; Mon, 30 Apr 2007 19:04:29 -0400 5, 27 -- Received: from EMLabServices ([24.255.213.54]) 5, 27 -- by eastrmimpo02.cox.net with bizsmtp 5, 27 -- id tb4K1W00d1AzDvc0000000; Mon, 30 Apr 2007 19:04:28 -0400 5, 27 -- Message-ID: {059201c78b84$62927da0$6400a8c0-at-EMLabServices} 5, 27 -- From: "EM Lab Services" {emlabservices-at-cox.net} 5, 27 -- To: {Microscopy-at-microscopy.com} 5, 27 -- Subject: [Microscopy] Re: Diffusion Pmp cooling water 5, 27 -- Date: Mon, 30 Apr 2007 18:05:09 -0600 5, 27 -- MIME-Version: 1.0 5, 27 -- Content-Type: text/plain; 5, 27 -- format=flowed; 5, 27 -- charset="iso-8859-1"; 5, 27 -- reply-type=original 5, 27 -- Content-Transfer-Encoding: 7bit 5, 27 -- X-Priority: 3 5, 27 -- X-MSMail-Priority: Normal 5, 27 -- X-Mailer: Microsoft Outlook Express 6.00.2900.3028 5, 27 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.3028 ==============================End of - Headers==============================
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
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