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From: Philip.Koeck-at-biosci.ki.se
Date: Mon, 2 Jul 2007 02:39:43 -0500
Subject: [Microscopy] RE: kV or keV?

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Don't understand the response below. Everything I received on this topic came through the Listserver plus it seems that the original request for info was also to the Listserver and that responses are of interest to everyone.

Ted

----- Original Message ----
X-from: Jim Quinn www.matscieng.sunysb.edu}
To: drteddunne-at-yahoo.com
Sent: Saturday, June 30, 2007 11:52:50 PM

I heartily agree! Make them suffer!)

-----Original Message-----


Except that students in my microprobe course who try to use these
terms interchangeably loose points. The same happens in the TEM
course taught at our university. It is the same difference between
potential and potential energy -- subtle but non-trivial.

Best,
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
==========================


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7, 25 -- To: {frah0010-at-umn.edu} , {microscopy-at-microscopy.com}
7, 25 -- Subject: RE: [Microscopy] kV or keV?
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From: jacques.faerber-at-ipcms.u-strasbg.fr
Date: Mon, 2 Jul 2007 04:15:52 -0500
Subject: [Microscopy] kV or keV?

Contents Retrieved from Microscopy Listserver Archives
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I would loose much points in your courses !! Is it necessary to be so
rigorous ?

As soon as one put an electrostatic element in an electron optical
device, one have to handel with eV. An electrostatic lens changes the
kinetcic energy of an electron, and SEM and TEM have much such elements
(wehnelt, extraction (FE) and accelerating anodes, SE or BE detectors,
electron filtering).

From a purist point of view, it could be said that only the electronic
engeneer is concerned with the kV notation, in the design/mainteance of
the HV circuit. The user has more to deal with the kinetic energy.

In the practise, it seems that biologists and mineralogist use more kV,
and physicists use more keV. None is wrong !

So be kind with your studens, there is much enough stuff which is
difficult to understand !

J. Faerber
IPCMS-GSI
(Institut de Physique et Chimie des Matériaux de Strasbourg
Groupe Surface et Interfaces)
23, rue de Loess ; BP43
67034 Strasbourg CEDEX 2
France

Tel 00 33(0)3 88 10 71 01
Fax 00 33(0)3 88 10 72 48
E-mail Jacques.Faerber-at-ipcms.u-strasbg.fr



Philip.Koeck-at-biosci.ki.se a écrit :
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} I heartily agree! Make them suffer!)
}
} -----Original Message-----
}
}
} Except that students in my microprobe course who try to use these
} terms interchangeably loose points. The same happens in the TEM
} course taught at our university. It is the same difference between
} potential and potential energy -- subtle but non-trivial.
}
} Best,
} 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
} ==========================
}
}
} ==============================Original Headers==============================
} 7, 25 -- From Philip.Koeck-at-biosci.ki.se Mon Jul 2 02:39:43 2007
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} 7, 25 -- Subject: RE: [Microscopy] kV or keV?
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From: anne.vaahtokari-at-helsinki.fi
Date: Mon, 2 Jul 2007 07:16:16 -0500
Subject: [Microscopy] Virus infected microscope samples

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html

Hello All,

Our imaging core facility needs to have rules of how to handle viral-
infected (replication competent) specimen in the microscopy room. I cannot
simply forbid such samples as people need virus infection in order to
efficiently express fluorescent proteins.

In the virus core facility, Barrydin (cocospropylendiaminguanidindiacetat,
didecyloxyethylmethylammoniumpropionat; Pan Biotech) is used for
disinfection. Does anybody know if this agent can safely be used for
cleaning microscopes in case of a spill? How about objectives? As both
retro/lenti- and adenoviruses are being used, ethanol is not enough.

Perhaps somebody has already devised guidelines for handling infectious
microscope samples and is willing to share them?

Regards,
Anne
--
Anne Vaahtokari, Ph.D
Imaging Coordinator
Molecular Imaging Unit
Biomedicum Helsinki
P.O. Box 63 (Haartmaninkatu 8)
FIN-00014 University of Helsinki
Finland
Tel: +358-9-191 25579
Fax: +358-9-191 25554
E-mail: anne.vaahtokari-at-helsinki.fi
MIU web site: www.miu.helsinki.fi




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From: nizets2-at-yahoo.com
Date: Mon, 2 Jul 2007 07:58:57 -0500
Subject: [Microscopy] Re: Virus infected microscope samples

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html

Hello!

I suppose you are talking about light microscopes.
I can't answer directly to your question, but I would
definitively avoid any contact with the objectives, it
would make things a lot easier to handle. As far as
possible use coverslips and desinfect them carefully
before microscoping. Handling replication-competent
human retroviruses otherwise (outside of a proper
hood) is unconsciousness for me. And, if not stored,
the samples must be autoclaved afterwards of course.

Gluraldehydes and formaldehydes are also desinfecting
agents, I don't think they could damade objectives but
they need hours to work efficiently and I am not sure
that soaking objectives for hours in aqueous solutions
is very good.

Now an alternative to chemical decontamination is the
possibility to UV-B decontaminate. I think UV-B kill
also virus.

Probably the best partner to discuss the sentivity of
the objectives to various chemicals is the
constructor.

Best regards,

Stephane




--- anne.vaahtokari-at-helsinki.fi wrote:

}
}
}
}
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} Hello All,
}
} Our imaging core facility needs to have rules of how
} to handle viral-
} infected (replication competent) specimen in the
} microscopy room. I cannot
} simply forbid such samples as people need virus
} infection in order to
} efficiently express fluorescent proteins.
}
} In the virus core facility, Barrydin
} (cocospropylendiaminguanidindiacetat,
} didecyloxyethylmethylammoniumpropionat; Pan Biotech)
} is used for
} disinfection. Does anybody know if this agent can
} safely be used for
} cleaning microscopes in case of a spill? How about
} objectives? As both
} retro/lenti- and adenoviruses are being used,
} ethanol is not enough.
}
} Perhaps somebody has already devised guidelines for
} handling infectious
} microscope samples and is willing to share them?
}
} Regards,
} Anne
} --
} Anne Vaahtokari, Ph.D
} Imaging Coordinator
} Molecular Imaging Unit
} Biomedicum Helsinki
} P.O. Box 63 (Haartmaninkatu 8)
} FIN-00014 University of Helsinki
} Finland
} Tel: +358-9-191 25579
} Fax: +358-9-191 25554
} E-mail: anne.vaahtokari-at-helsinki.fi
} MIU web site: www.miu.helsinki.fi
}
}
}
}
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} {anne.vaahtokari-at-helsinki.fi}
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From: frah0010-at-umn.edu
Date: Mon, 2 Jul 2007 08:31:11 -0500
Subject: [Microscopy] kV or keV?

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html

Philip Koeck wrote:

} I heartily agree! Make them suffer!)

Jacques Faerber wrote:

} I would loose much points in your courses!! Is it necessary to be
} so rigorous? ... So be kind with your studens, there is much enough
} stuff which is difficult to understand!


Jacques, Philip, and everyone,

I do not grade such details just to be nasty or make the students
suffer (although they might disagree). Many of these students will
go on to use our electron microprobe for their research, and they
will be presenting posters, giving conference talks, and submitting
papers to publish. If one of the students mentions using "an
accelerating voltage of 15 keV" or observing "X-ray peaks at 4.57 and
7.14 kV," there will be reviewers or audience members who notice such
mistakes and may start to doubt the student's understanding of the
analytical technique and, consequently, their results and
conclusions. I grade for such details so that the students to do
make the same mistakes later and embarrass themselves (and me as
their teacher) when it really matters. Hopefully they understand
that my rigorous grading is in their best interests -- I think that
most of them do, and I hear far more grumbling about the essays that
I make them write on quizzes and exams.

Best,
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

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From: anne.vaahtokari-at-helsinki.fi
Date: Mon, 2 Jul 2007 08:38:27 -0500
Subject: [Microscopy] Re: Virus infected microscope samples

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html

Hi,

A correction and clarification for my posting: the virus infected samples
are NOT replication competent, and they are for live cell imaging (both
wide-field and confocal microscopes are used).

Anne
--
Anne Vaahtokari, Ph.D
Imaging Coordinator
Molecular Imaging Unit
Biomedicum Helsinki
P.O. Box 63 (Haartmaninkatu 8)
FIN-00014 University of Helsinki
Finland
Tel: +358-9-191 25579
Fax: +358-9-191 25554
E-mail: anne.vaahtokari-at-helsinki.fi
MIU web site: www.miu.helsinki.fi




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From: jacques.faerber-at-ipcms.u-strasbg.fr
Date: Tue, 3 Jul 2007 02:52:36 -0500
Subject: [Microscopy] kV or keV?

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html

Hi Ellery and all

Of coarse, I agree that students have to learn the right words/units to
express the right concepts. But what I wanted to point out, is that, in
the description of the SEM/TEM working principle, the two concepts have
their place.

I understand than one accept, in the following sentances, either :

"We have observed sample xyz with a an accelerating voltage of 15 kV"
or :
"We have observed sample xyz with a an electron beam energy set to
15 keV"

and not the contrary or a unprecise mixing of the two formulations. But
one cannot reject the formulation using the energy concept. I don't
speak here from x-rays, only from the electrons in the microscope.

In my former mail I mentionned the electrostatic elements of the
microscope. In the electron/matter interaction too, it's the energy
aspect which plays a role, not the voltage difference used to
accelerate/decelerate these electrons. In diffraction, the equivalence
is between wavelength and energy.

Jacques

J. Faerber
IPCMS-GSI
(Institut de Physique et Chimie des Matériaux de Strasbourg
Groupe Surface et Interfaces)
23, rue de Loess ; BP43
67034 Strasbourg CEDEX 2
France

Tel 00 33(0)3 88 10 71 01
Fax 00 33(0)3 88 10 72 48
E-mail Jacques.Faerber-at-ipcms.u-strasbg.fr



Ellery Frahm a écrit :
} Philip Koeck wrote:
}
} } I heartily agree! Make them suffer!)
}
} Jacques Faerber wrote:
}
} } I would loose much points in your courses!! Is it necessary to be so
} } rigorous? ... So be kind with your studens, there is much enough
} } stuff which is difficult to understand!
}
}
} Jacques, Philip, and everyone,
}
} I do not grade such details just to be nasty or make the students
} suffer (although they might disagree). Many of these students will go
} on to use our electron microprobe for their research, and they will be
} presenting posters, giving conference talks, and submitting papers to
} publish. If one of the students mentions using "an accelerating
} voltage of 15 keV" or observing "X-ray peaks at 4.57 and 7.14 kV,"
} there will be reviewers or audience members who notice such mistakes
} and may start to doubt the student's understanding of the analytical
} technique and, consequently, their results and conclusions. I grade
} for such details so that the students to do make the same mistakes
} later and embarrass themselves (and me as their teacher) when it
} really matters. Hopefully they understand that my rigorous grading is
} in their best interests -- I think that most of them do, and I hear
} far more grumbling about the essays that I make them write on quizzes
} and exams.
}
} Best,
} 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

==============================Original Headers==============================
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12, 29 -- Subject: Re: [Microscopy] RE: kV or keV?
12, 29 -- References: {200707020747.l627lmVV012877-at-ns.microscopy.com} {4FDB017F-1557-43CB-8E0E-64C7E4A36319-at-umn.edu}
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From: catherine.bougerol-at-cea.fr
Date: Tue, 3 Jul 2007 07:12:51 -0500
Subject: [Microscopy] viaWWW: SEM of particles on quartz

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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
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Email: catherine.bougerol-at-cea.fr
Name: bougerol

Organization: CNRS

Title-Subject: [Filtered] SEM of particles on quartz

Question: I have a sample consisting of metallic particles (about 15nm diameter) deposited on a quartz or glass slide. I wonder if and how it is possible to get an image by SEM, with a FEG microscope.

---------------------------------------------------------------------------

==============================Original Headers==============================
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From: nizets2-at-yahoo.com
Date: Tue, 3 Jul 2007 07:16:09 -0500
Subject: [Microscopy] Re: SEM: Wet SEM capsule

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html

Thanks for the infos!
It looks very much like a revolution to me! (if it
really works as it is presented ;-))
One can expected a deep penetration of the beam into
cells (but a larger dispersion too of course).

The only drawback I noticed: in the FAQ, they do not
answer to their own question "What is the price of the
QX-202C capsules?" and this is usually bad sign :-D

Regards

Stephane


--- kraftpiano-at-gmail.com wrote:

}
}
}
}
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}
} I was recently perusing one of the supply catalogs
} for SEM supplies,
} and I found a wet-cell capsule that has an "Electron
} transparent and
} vacuum tight window."
}
(http://www.emsdiasum.com/microscopy/products/sem/wet/technical.aspx)
} I was just wondering if anybody has used these, and
} what kind of
} results they are getting.
}
} The second part of my question is a bit more
} abstract. What quality
} makes something electron transparent, and what
} materials are electron
} transparent?
}
} --Justin A. Kraft
}
} ==============================Original
} Headers==============================
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} 09:50:21 2007
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From: frah0010-at-umn.edu
Date: Tue, 3 Jul 2007 08:35:55 -0500
Subject: [Microscopy] Re: kV or keV?

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html

Hi Jacques and all,

Oh, yes, I absolutely agree that both units/concepts (potential/
voltage and energy) have their place in electron microscopy, just as
energy and wavelength are both valid ways to think about X-ray
analysis -- I didn't mean to suggest otherwise. I simply meant that
students must use the proper terminology and units together and in
the most appropriate situations, and we agree on that too. Like I
said in the previous message, what is unacceptable is, for example,
saying "an accelerating voltage of 15 keV" or describing X-ray peaks
in units of kV. But students should be familiar with both ways of
thinking about the technique.

Best,
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


On Jul 3, 2007, at 3:00 AM, jacques.faerber-at-ipcms.u-strasbg.fr wrote:

} Hi Ellery and all
}
} Of coarse, I agree that students have to learn the right words/
} units to
} express the right concepts. But what I wanted to point out, is
} that, in
} the description of the SEM/TEM working principle, the two concepts
} have
} their place.
}
} I understand than one accept, in the following sentances, either :
}
} "We have observed sample xyz with a an accelerating voltage of
} 15 kV"
} or :
} "We have observed sample xyz with a an electron beam energy
} set to
} 15 keV"
}
} and not the contrary or a unprecise mixing of the two formulations.
} But
} one cannot reject the formulation using the energy concept. I don't
} speak here from x-rays, only from the electrons in the microscope.
}
} In my former mail I mentionned the electrostatic elements of the
} microscope. In the electron/matter interaction too, it's the energy
} aspect which plays a role, not the voltage difference used to
} accelerate/decelerate these electrons. In diffraction, the equivalence
} is between wavelength and energy.
}
} Jacques
}
} J. Faerber
} IPCMS-GSI
} (Institut de Physique et Chimie des Matériaux de Strasbourg
} Groupe Surface et Interfaces)
} 23, rue de Loess ; BP43
} 67034 Strasbourg CEDEX 2
} France
}
} Tel 00 33(0)3 88 10 71 01
} Fax 00 33(0)3 88 10 72 48
} E-mail Jacques.Faerber-at-ipcms.u-strasbg.fr
}
}
}
} Ellery Frahm a écrit :
} } Philip Koeck wrote:
} }
} } } I heartily agree! Make them suffer!)
} }
} } Jacques Faerber wrote:
} }
} } } I would loose much points in your courses!! Is it necessary to
} } } be so
} } } rigorous? ... So be kind with your studens, there is much enough
} } } stuff which is difficult to understand!
} }
} }
} } Jacques, Philip, and everyone,
} }
} } I do not grade such details just to be nasty or make the students
} } suffer (although they might disagree). Many of these students
} } will go
} } on to use our electron microprobe for their research, and they
} } will be
} } presenting posters, giving conference talks, and submitting papers to
} } publish. If one of the students mentions using "an accelerating
} } voltage of 15 keV" or observing "X-ray peaks at 4.57 and 7.14 kV,"
} } there will be reviewers or audience members who notice such mistakes
} } and may start to doubt the student's understanding of the analytical
} } technique and, consequently, their results and conclusions. I grade
} } for such details so that the students to do make the same mistakes
} } later and embarrass themselves (and me as their teacher) when it
} } really matters. Hopefully they understand that my rigorous
} } grading is
} } in their best interests -- I think that most of them do, and I hear
} } far more grumbling about the essays that I make them write on quizzes
} } and exams.
} }
} } Best,
} } 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
}
} ==============================Original
} Headers==============================
} 12, 29 -- From jacques.faerber-at-ipcms.u-strasbg.fr Tue Jul 3
} 02:52:35 2007
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} Microscopy-at-microscopy.com
} 12, 29 -- Subject: Re: [Microscopy] RE: kV or keV?
} 12, 29 -- References:
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} {4FDB017F-1557-43CB-8E0E-64C7E4A36319-at-umn.edu}
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From: randy-nessler-at-uiowa.edu
Date: Tue, 3 Jul 2007 12:44:41 -0500
Subject: [Microscopy] Micro IR and Ramam systems

Contents Retrieved from Microscopy Listserver Archives
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I'm posting for a colleague who is interested in finding various vendors
of the technology listed below. Obviously, vendors are encouraged to
reply. Anybody have vendor/system recommendations?
Randy


Who manufactures micro IR and Raman systems (various combinations of
NIR, mid IR, far IR, Raman) for our existing light microscopes? It
shouldn't necessarily be a system that is mounted right on top of the
microscope (as in Smiths detection case). There are bench top systems
that can be coupled to a light microscope as well.

-----------------------------------------------------------
Jonas Baltrusaitis, Ph.D.
Postdoctorate Research Fellow
Department of Chemistry
and
Central Microscopy Research Facility
85 EMRB
University of Iowa
Iowa City, IA 52242
phone: 319-335-8142


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From: damondewsy-at-yahoo.com
Date: Tue, 3 Jul 2007 21:17:01 -0500
Subject: [Microscopy] viaWWW: Phillips EM201 Installations manual

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This Question/Comment was submitted to the Microscopy Listserver
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Email: damondewsy-at-yahoo.com
Name: Damon Dewsbury

Organization: University of Toronto

Title-Subject: [Filtered] Phillips EM201 Installations manual

Question: Hello,
I'm trying to reassemble a Phillips EM201 TEM. I have the other 3 parts of the manual (including Pre-installation). I was wondering if anyone might be able to tell me who I might get this manual from or if someone might have a pdf copy of it. Thanks.

Damon Dewsbury
MSc candidate
UofT- Dept. of EEB
damondewsy-at-yahoo.com

---------------------------------------------------------------------------

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From: Jane.LaGoy-at-Bodycote.com
Date: Thu, 5 Jul 2007 08:46:37 -0500
Subject: [Microscopy] Problem with EDS signal on SEM

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


Hello microscopists,

I'm hoping someone can explain a strange phenomenon that's occurred with
my EDAX EDS - JEOL SEM system. Actually, two things: 1) There is a
huge 'noise' peak at the far left of the EDS energy spectrum that
doesn't correspond to elements present, and 2) The count rate (CPS)
while gathering a spectrum is abnormally high, but the dead time (DT%)
is very low - this is the opposite of what normally occurs. It happens
in both secondary and backscattered electron modes. I'm looking at
metal samples, and I gathered a spectrum from the aluminum sample holder
to make sure what I was seeing was real. Unfortunately, it is. Any
ideas of what's going on, or what troubleshooting I should try, will be
greatly appreciated.

Thank you,

Jane L. LaGoy
Laboratory Services Manager/
Development Engineer
Bodycote North America
155 River Street
Andover, MA 01810
978-470-1620 x450
FAX: 978-475-2951
jane.lagoy-at-bodycote.com
The only people to get even with are those who have helped you.

This Email and all attachments hereto, if any, are covered by the provisions of the U.S. Electronic Communications Privacy Act. Additionally, all contents of this communication and all attachments hereto shall be deemed to be confidential and the contents thereof are proprietary to Bodycote Thermal Processing, Inc. ("Bodycote"). If this communication has been sent to you in error, please reply to the sender that you received it and then delete the message immediately. Any dissemination, distribution, copying or reproduction of this message other than by its intended recipient is strictly prohibited.



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From: NWWhite-at-bwxt.com
Date: Thu, 5 Jul 2007 09:37:14 -0500
Subject: [Microscopy] Problem with EDS signal on SEM

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html

Hello Jane,

A number of possibilities come to mind, assuming you have a UTW, LN2
cooled detector. Imaging mode (SEI vs. BSE) should have no bearing on
the EDS noise unless (remote possibility) a detector is generating
electrical noise.

1. There could be a problem with the pulse processor. Low-end
noise is suppressed when working properly.

2a. LN2 boiling due to ice crystals in dewar.

2b. LN2 boiling due to detector vacuum leak (typically from bad UTW)


3. Extraneous light source. Usually from a vacuum gauge or the LED
illumination from a chamber (TV) scope.

4. Detector bias overvoltage, damaging detector/preamp.

5. Damaged detector from warm-up with bias applied.

6. Missing electron trap on end of detector.

7. Ground loop in signal wires. E.G: broken cable shield,
disconnected ground wires. (Has the wiring layout changed or any
electrically noisy new equipment been installed anywhere nearby?)

...All that comes to mind at the moment. Good Luck!

Woody

NW (Woody) White Jr
BWXT Services
Lynchburg, VA
http://www.bwxt.com/operations/semlab.html


-----Original Message-----
X-from: Jane.LaGoy-at-Bodycote.com [mailto:Jane.LaGoy-at-Bodycote.com]
Sent: Thursday, July 05, 2007 9:48 AM
To: White, Woody N.


Hello microscopists,

I'm hoping someone can explain a strange phenomenon that's occurred with
my EDAX EDS - JEOL SEM system. Actually, two things: 1) There is a
huge 'noise' peak at the far left of the EDS energy spectrum that
doesn't correspond to elements present, and 2) The count rate (CPS)
while gathering a spectrum is abnormally high, but the dead time (DT%)
is very low - this is the opposite of what normally occurs. It happens
in both secondary and backscattered electron modes. I'm looking at
metal samples, and I gathered a spectrum from the aluminum sample holder
to make sure what I was seeing was real. Unfortunately, it is. Any
ideas of what's going on, or what troubleshooting I should try, will be
greatly appreciated.

Thank you,

Jane L. LaGoy
Laboratory Services Manager/
Development Engineer
Bodycote North America
155 River Street
Andover, MA 01810
978-470-1620 x450
FAX: 978-475-2951
jane.lagoy-at-bodycote.com
The only people to get even with are those who have helped you.

This Email and all attachments hereto, if any, are covered by the
provisions of the U.S. Electronic Communications Privacy Act.
Additionally, all contents of this communication and all attachments
hereto shall be deemed to be confidential and the contents thereof are
proprietary to Bodycote Thermal Processing, Inc. ("Bodycote"). If this
communication has been sent to you in error, please reply to the sender
that you received it and then delete the message immediately. Any
dissemination, distribution, copying or reproduction of this message
other than by its intended recipient is strictly prohibited.



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From: smalinskas-at-yahoo.com
Date: Thu, 5 Jul 2007 12:34:29 -0500
Subject: [Microscopy] Re: Micro IR and Ramam systems

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html

Randy,

My colleague is very knowledgable on Raman systems.
He suggested looking into Kaiser Optical Systems
(http://www.kosi.com/).

Kestutis Smalinskas, P.E.
Sr. Metallurgist
SKF USA
Plymouth, Michigan
(734) 414-6862

--- randy-nessler-at-uiowa.edu wrote:
} I'm posting for a colleague who is interested in
} finding various vendors
} of the technology listed below. Obviously, vendors
} are encouraged to
} reply. Anybody have vendor/system recommendations?
} Randy
}
}
} Who manufactures micro IR and Raman systems (various
} combinations of
} NIR, mid IR, far IR, Raman) for our existing light
} microscopes? It
} shouldn't necessarily be a system that is mounted
} right on top of the
} microscope (as in Smiths detection case). There are
} bench top systems
} that can be coupled to a light microscope as well.
}
}
-----------------------------------------------------------
} Jonas Baltrusaitis, Ph.D.
} Postdoctorate Research Fellow
} Department of Chemistry
} and
} Central Microscopy Research Facility
} 85 EMRB
} University of Iowa
} Iowa City, IA 52242
} phone: 319-335-8142
}
}
} ==============================Original
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} 12:44:41 2007
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From: gary-at-gaugler.com
Date: Thu, 5 Jul 2007 21:09:38 -0500
Subject: [Microscopy] Problem with EDS signal on SEM

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html

At 06:38 AM 7/5/2007, you wrote:




} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America

No. This would produce high cps and 100% DT.


} 4. Detector bias overvoltage, damaging detector/preamp.

Unlikely.


} 5. Damaged detector from warm-up with bias applied.

Also unlikely.


} 6. Missing electron trap on end of detector.

What? It just fell off? Also not likely.


} 7. Ground loop in signal wires. E.G: broken cable shield,
} disconnected ground wires. (Has the wiring layout changed or any
} electrically noisy new equipment been installed anywhere nearby?)

Did something change in the wiring or layout? This can have
a negative impact.



} ...All that comes to mind at the moment. Good Luck!
}
} Woody
}
} NW (Woody) White Jr
} BWXT Services
} Lynchburg, VA
} http://www.bwxt.com/operations/semlab.html


Peaks at low eV can be due to noise and/or shifts
in BLM. Change to a lower uS time constant and
see what happens. If the same condition persists,
then you have a hardware problem. Hopefully, you
have a maintenance contract with EDAX. That should
fix it.

gary g.




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27, 20 -- To: NWWhite-at-bwxt.com
27, 20 -- From: Gary Gaugler {gary-at-gaugler.com}
27, 20 -- Subject: Re: [Microscopy] RE: Problem with EDS signal on SEM
27, 20 -- Cc: MSA listserver {microscopy-at-microscopy.com}
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From: dsherman-at-purdue.edu
Date: Thu, 5 Jul 2007 21:10:26 -0500
Subject: [Microscopy] Sorvall JB-4 knife breaker

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html

Listers,

We have a Sorvall knife breaker used to break the thicker "Ralph knives" for
use with a Sorvall JB-4 microtome. It is having problems so I am looking
for a manual that may give some use instructions and schematics.

Alternatively we may need to find someone to repair it. I would appreciate
hearing from or about anyone who works on these instruments.

I might also be interested in buying a replacement if anyone has one sitting
around not being used.

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



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From: youngwk-at-snu.ac.kr
Date: Sun, 8 Jul 2007 20:52:07 -0500
Subject: [Microscopy] EDS switching in F20

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html

Listers,

We have problems of switching EDS from the embedded to the stand-alone
operation in our F20 STEM.
We used to use EDS as a embedded system through TIA interface, and we now
want to use switching capability in the TIA software, which provides a
checkbox to switch from the embedded to stand-alone, external EDS collection
system. The checkbox, however, did not work. To switch, we have to change
parameters in engineering mode then restart the control computer, which
means you have to begin from the emission start.
Is there anyone who are using the EDS switching function in the TIA
software? Or information about any method other than restarting the computer
would be appreciated.


Young-Woon Kim
Associate Professor
Seoul National University
School of Materials Science and Engineering
Tel) +82-2-880-7977
Fax) +82-2-883-8197
e-mail) youngwk-at-snu.ac.kr






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From: kraftpiano-at-gmail.com
Date: Sun, 8 Jul 2007 21:16:42 -0500
Subject: [Microscopy] SEM Donation, Round 2, and a part request.

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html

Well, I've received two offers from promising schools locally who
would be willing to take on both me and my lab. They are high school
programs, so I will be able to pick up right where I left off on
developing my microscopy course.

I have acquired another donation of an SEM, this time a JEOL JSM-35C.
I'll post pictures and the website to them when it gets here. This
time, I've got an EDS system with it, so I might actually be able to
do some imaging!

Now, the request. I also have a Topcon/ISI ABT-SX40A that needs a
little help. I have reduced the problem down to a fault in the HV
stablizer board, part number N110HC01. If anybody has this part
hanging around, I would appreciate it.

The second option is for me to go ahead and digitize the scope. I
know what the base voltages are going to the HV tank, and I have been
able to control the emission current manually with a DC power supply
injecting voltage into the circuit, but I don't have a D/A conversion
board that would be able to control the outputs. Any suggestions on
cheap ones?

Thanks,

Justin A. Kraft

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From: paul_hazelton-at-umanitoba.ca
Date: Mon, 9 Jul 2007 08:50:06 -0500
Subject: [Microscopy] specimen holder repair

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html

As time goes by, and equipment gets older, the need to repair gets
greater. Does anyone have a good source for repairing the wire side bar
for the old Philips specimen holders?

help would be very greatly appreciated.

Paul

Paul R. Hazelton, PhD
Electron Microscope Unit
University of Manitoba
Department of Medical Microbiology
531 Basic Medical Sciences Building
730 William Avenue
Winnipeg, Manitoba, Canada, R3E 0W3
e-mail: paul_hazelton-at-umanitoba.ca
Phone:204-789-3313
Pager:204-931-9354
Cell:204-781-1502
Fax:204-789-3926


==============================Original Headers==============================
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From: colijn.1-at-osu.edu
Date: Mon, 9 Jul 2007 09:52:35 -0500
Subject: [Microscopy] Re: specimen holder repair

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html

Hi Paul,

We've been sending our holders to Tom Schmeltzer for repair and been
quite happy with his work.

Tom Schmeltzer
TGS Technologies
900 Glenwood Ct.
Cranberry Township, PA 16066
(724) 453-3865

...just a satisfied customer...

Cheers,
Henk


At 09:52 AM 07/09/07, paul_hazelton-at-umanitoba.ca wrote:



} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America

Hendrik O. Colijn colijn.1-at-osu.edu
Campus Electron Optics Facility Ohio State University
(614) 292-0674 http://www.ceof.ohio-state.edu
Time is that quality of nature which keeps events from happening all
at once. Lately it doesn't seem to be working.


==============================Original Headers==============================
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12, 26 -- Date: Mon, 09 Jul 2007 10:53:32 -0400
12, 26 -- From: "Hendrik O. Colijn" {colijn.1-at-osu.edu}
12, 26 -- Subject: Re: [Microscopy] specimen holder repair
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From: Geoffrey_Williams-at-brown.edu
Date: Mon, 9 Jul 2007 10:28:10 -0500
Subject: [Microscopy] Re: specimen holder repair

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html

I will publicly second the recommendation for Tom's work. The holder
for a CM10 was repaired beautifully, in a timely manner at a reasonable
cost.

Again, no connection, other than a satisfied customer.

Geoff Williams
Leduc Bioimaging Facility Manager
Brown University

http://www.brown.edu/Facilities/Leduc_Bioimaging_Facility/

-----Original Message-----
X-from: colijn.1-at-osu.edu [mailto:colijn.1-at-osu.edu]
Sent: Monday, July 09, 2007 10:56 AM
To: Williams, Geoffrey

Hi Paul,

We've been sending our holders to Tom Schmeltzer for repair and been
quite happy with his work.

Tom Schmeltzer
TGS Technologies
900 Glenwood Ct.
Cranberry Township, PA 16066
(724) 453-3865

...just a satisfied customer...

Cheers,
Henk


At 09:52 AM 07/09/07, paul_hazelton-at-umanitoba.ca wrote:



} -----------------------------------------------------------------------
-----
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of
America

Hendrik O. Colijn colijn.1-at-osu.edu
Campus Electron Optics Facility Ohio State University
(614) 292-0674 http://www.ceof.ohio-state.edu
Time is that quality of nature which keeps events from happening all
at once. Lately it doesn't seem to be working.


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12, 26 -- Subject: Re: [Microscopy] specimen holder repair
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==============================Original Headers==============================
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From: jcai-at-nanostellar.com
Date: Mon, 9 Jul 2007 12:23:54 -0500
Subject: [Microscopy] commercial lab with state of the art FEG-STEM

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html

Dear All,

I am looking for commercial labs equipped with good FEG-STEM, which is
able to do EDS analysis.  The STEM probe size is preferred to be ~0.5-1
nm, and it would be good to have high angle annular dark filed image
available.

If you have any information about it, please let me know.  Your help
will be highly appreciated.


Juan



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From: TindallR-at-missouri.edu
Date: Mon, 9 Jul 2007 15:23:28 -0500
Subject: [Microscopy] Summary: SEM/TEM--Carbon nanoparticles in tissue

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Here at long last is my promised summary of replies to my question about how to image carbon nanoparticles that have been taken up by organisms into tissue. Hope I didn't miss anybody.

Many thanks again to all who replied.



X-from Mhairi Gass: "A colleague has brought my attention to your question on visualising carbon nanos in tissue. I have been working on a similar project with Alex Porter (Cambridge University) where we have been exploiting a range of techniques to visualise C60 and SWNTs in cellular materials. We have found that staining the specimens can often add confusion, but is useful to identify the different parts of a cell. As for differentiating between the carbonaceous materials we have found that a ratio of plasmon energies in EFTEM works very well but this must be on unstained sections (i.e. EFTEM images at 26eV and 22eV ish, or even using the pi to pi* transition at 6eV for crystalline materials). I have attached a paper that was published earlier this year which may be of interest. Also HAADF imaging can enhance the contrast from the crystalline material as it has a higher density. We currently have a paper in review on unstained imaging at high resolution which involves preparing much thinner samples than are traditionally used for biological samples to minimise the 3D information, they are not easy to work with but the resolution is much improved." (The paper attached was: Visualizing the Uptake of C60 to the Cytoplasm and Nucleus of Human
Monocyte-Derived Macrophage Cells Using Energy-Filtered Transmission Electron Microscopy and Electron Tomography. Alexandrae Porter, Mhairi Gass, Karin Muller, Jeremyn Skepper, Paul Midgley, and Mark Welland. Environ. Sci. Technol. 2007, 41, 3012-3017. RT)
*************************

X-from Hendrik Colijn: "I don't think the nano-thingys will take much stain. You may have better luck staining the tissue and working with a sort of "negative stain" If the nanothingys are crystalline enough, you may be able to do dark field imaging."
**************************

X-from Ephram Mark Shizgal: "Randy, I would love to give those sections a look under our low voltage TEM (see http://www.lv-em.com/ RT--no connection with me, etc., but interesting information) to see if the extra contrast low kV provides enhances the nanomaterials. How nano are they? I know that you guys don't have the LVEM5, but if it was something you'd want to try we would be happy to look at them at our applications lab. Let me know."
********************************

X-from Phil Oshel: "The only way I see is to incorporate C-14 into the nanobits when they're made, then use old-fashioned radio-immuno techniques. Autoradiography. Don't see how this would help with SEM, but would work for TEM and LM."
********************************

X-from Barbara Foster: "This is another one of those "TOMO" applications.

As mentioned earlier, I have a movie made by Dr.deWith of the Dutch Polymer Institute (TU/E) showing the distribution of carbon nanotubes in epoxy. Because the AFM uses local differences in elasticity, there is none of the CNT in C-based tissue problem described below. Again, although I am no longer involved in this project, I am happy to forward a copy of the film to anyone who is interested. (It will come via www.YouSendIt.com, because of the large file size).

Dr. Anton Efimov of NT-MDT is the developer of this technique and is usually willing to run samples, schedule permitting. See his email in the CC: above.

NTA is no longer NT-MDT's agent here in the US. If you are interested in following this further, I recommend that you contact Dr. Yuri Bobrov, who is now with ABeam (Dealer principle, Dr. Sergey Babin - see email above).

As always, if there are further questions, please don't hesitate to call/email.

(Note: MME is no longer involved with this vendor)."
***********************************

X-from Petra Rudolf: "Bruce Weisman (weisman-at-rice.edu) has very nicely shown that it is possible to identify the type of CNT inside tissue from fluorescence spectroscopy - one can actually see fluorescence from a single tube and from the frequency identify what tube it is.He has also shown how to do statistics of the distribution of nanotubes in different types of tissue.
I heard his talk on this subject at the 2006 IWEPNM in Kirchberg, Austria."
*************************************

X-from Dale Callaham: "I have looked at carbon nanotubes a bit. I have looked at them applied to thin carbon films - no neg stain necessary - and they stand out well - very easy to image - especially the multiwall ones. I have also sectioned them in epoxy and melamine resins and it was no problem to see them easily in normal 60nm sections. In the presence of complex tissue it could be more difficult to see them. But the cells may react in some way to partition them and make them easy to see?

And I have seen many different shapes/lengths. The particular sample from a source is fairly consistent - I have never purchased so I don't know what people see about what they get, but they are often surprised when I show them the images. I went into it thinking that I was going to see little soda straws, but I have seen nearly any conformation you can imagine. Some disperse easily but I had some that took much sonication to disperse, and just a little swirl and they came out of suspension; those were the long curly ones - they tangle easily when swirled. So get a sample from your associates and see what you are dealing with."
********************************

X-from Stephane Nizet: "Been not involved in carbon nanotube research I dont know exactly how dense they are under the beam of a TEM. However I don't think you can compare the density of the carbon present in a tissue with the density of carbon in nanotubes. The difference MUST be visible.
I wouldn't modify the nanotubes themselves because you will definitely modify their behaviour in an uncontrolled manner.
Now here is what I would do:
I would incubate cell monolayers with the nanoparticles and flat embed them (control=cells incubated at 4°C, no uptake) after ferrocyanide-osmium post-fixation. I am pretty sure the cells will internalize the particles (take hepatocytes like HepG2 they are very effecient at uptaking and very nice to show). After sectionning, I would try different contrasting intensities and also without any contrasting at all!
This way you can define your technical parameters to obtain the best contrast possible in "control"
samples.

Finding these particles in organs is another story, requiring mainly eternities of patience and dedication.
But, if I may give my personal opinion, this is a very interesting study and probably a very rewarding one.
Few have had the heart to start it."
****************************************


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


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From: paul_hazelton-at-umanitoba.ca
Date: Wed, 11 Jul 2007 08:23:56 -0500
Subject: [Microscopy] Thanks - Specimen Holder Repair

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Thanks to all who responded to my problem with repair of specimen
holders. A number of very good sources for repair and replacement were
sent.

Once again, thanks

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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From: donovan-at-uoregon.edu
Date: Wed, 11 Jul 2007 14:29:28 -0500
Subject: [Microscopy] Instrument Service Engineer Position at the University of

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Position closes July 31st!

Electron/Ion Beam Instrument Service Engineer
The University of Oregon's Center for Advanced Materials
Characterization in Oregon (CAMCOR) is seeking applications for a
full time staff position to begin September 2007. A strong background
in maintaining, trouble shooting and upgrading electron/ion beam
instruments and associated high voltage, vacuum, mechanical and
electrical systems is required. Experience with x-ray diffraction
instrumentation is also desirable. Salary range $60K-90K
commensurate with experience.

This position will be located in the new Lorry Lokey Integrated
Science Laboratory, a state of the art nano and micro science
analytical instrument facility designed specifically for exceptional
nano-science performance. It will house the latest electron, ion and
x-ray beam instrumentation available including a Zeiss Ultra TFEM,
FEI Quanta 200 E-SEM, Cameca SX50 and SX100 microprobes, Ion-TOF
SIMS, Philips Auger Spectrometer, Philips TEM, Kratos XPS and various
assorted coaters, etchers, and other vacuum deposition systems.

The successful candidate with have a BS in a beam microscopy related
field and an extensive background in instrument field service with
significant practical experience troubleshooting high vacuum electron
and ion beam instrumentation at both the system and PC board levels.
Must be able to read and understand schematics for electronic
circuits and systems. The successful applicant will be involved in
modifying/improving instrumentation capabilities to enable the
equipment to more fully support unique research needs and will be
expected to work intimately with the scientific staff and research
faculty. We seek candidates with a demonstrated commitment to working
effectively with students, faculty and staff from diverse backgrounds.

Interested persons should send a resume with a detailed description
of work experience and skills, and arrange for two letters of
recommendation to be sent to: CAMCOR Instrument Engineer Search
Committee, 1253 University of Oregon, Eugene, OR, 97403-1253. To be
assured of full consideration, application materials must be received
by July 31, 2007, but the search will remain open until the position
is filled. For further information, contact John Donovan (donovan-at-uoregon.edu).

University of Oregon is an AA/EEO employer committed to cultural diversity.


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From: gas19-at-daimlerchrysler.com
Date: Wed, 11 Jul 2007 18:15:02 -0500
Subject: [Microscopy] viaWWW: Micro IR and Ramam systems

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Email: gas19-at-daimlerchrysler.com
Name: Gerald Shulke

Organization: DaimlerChrysler

Title-Subject: [Microscopy] Re: Micro IR and Ramam systems

Question: I just saw a presentation from Diane Allen from Bruker Optics. Check out the website brukeroptics.com.

Gerald Shulke
DaimlerChrysler Corporation

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From: WLehman-at-bu.edu
Date: Wed, 11 Jul 2007 18:15:29 -0500
Subject: [Microscopy] viaWWW: EM position available

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Email: WLehman-at-bu.edu
Name: Tori Hatch

Organization: Boston University

Title-Subject: [Filtered] EM position available

Question: POSITION AVAILABLE
Senior Electron Microscopy Technician

A Senior Electron Microscopy Technician is sought to perform duties in support of research work on the structure of actin filament complexes that (1) are associated with cardiac and skeletal muscle regulatory proteins to control muscle activity and (2) interact with smooth muscle actin-binding proteins to modulate the assembly of the smooth muscle cytoskeleton. Applicants must have experience in high-resolution EM work and first-rate facility with computer-assisted image analysis. The candidate should have a Bachelorís degree and several years of experience. Prior familiarity with preserving and recording macromolecular assemblies in negative stain and by cryo-EM methods would be invaluable. Experience in supervising graduate students and post-doctoral fellows would also be important.

Interested applicants should send their CV with three references to:

Dr. William Lehman
Professor of Physiology & Biophysics
Department of Physiology & Biophysics
Boston University School of Medicine
715 Albany Street
Boston, MA 02118

Or email:

wlehman-at-bu.edu

Dr. Lehmanís research program is summarized on his home page:

http://biophysics.bumc.bu.edu/faculty/lehman/index.html



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From: medvitz-at-bostwicklaboratories.com
Date: Wed, 11 Jul 2007 18:15:53 -0500
Subject: [Microscopy] viaWWW: Uranyl acetate disposal

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Email: medvitz-at-bostwicklaboratories.com
Name: Neil Medvitz

Organization: Bostwick Laboratories

Title-Subject: [Filtered] Uranyl acetate disposal

Question: Does anyone know of a company that will take my uranyl acetate & uranyl nitrate waste in the Richmond Virginia area?? As usual as soon as someone hears uranyl they freak out.

Thanks,
Neil Medvitz

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From: fittonro-at-luther.edu
Date: Thu, 12 Jul 2007 08:39:35 -0500
Subject: [Microscopy] viaWWW: Gatan 791 manual wanted

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Email: fittonro-at-luther.edu
Name: Robert Fitton

Organization: Luther College

Title-Subject: [Filtered] Gatan 791 manual wanted

Question: Greetings, I'm looking for someone who would be willing to make a copy of the Gatan 791 manual that includes installation instructions and send it to me either snail mail or pdf. I'm specifically interested in the pneumatic connections and psi specifications. Thanks! Robert Fitton/Luther College/Decorah, IA


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From: lamiller-at-uiuc.edu
Date: Thu, 12 Jul 2007 12:43:08 -0500
Subject: [Microscopy] Taking the gluteraldehyde out

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Dear Folks on the Microscopy List,


I have a researcher who wants to do immuno labeling, but the sample is
already in gluteraldehyde.

I've always gone on the premise that gluteraldehyde is permanent, and
always suggest 4% parafromaldehyde or parafromaldehyde with the barest
littlest amount of gluteraldehyde ( {1%).


I am not familiar with any retrieval protocols, do you know of any for
gluteraldehyde, where the Ag site actually is affected by the
gluteraldehyde?



Thanks,


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


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14, 17 -- From: Lou Ann Miller {lamiller-at-uiuc.edu}
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From: lamiller-at-uiuc.edu
Date: Thu, 12 Jul 2007 13:04:40 -0500
Subject: [Microscopy] Re: Taking the gluteraldehyde out

Contents Retrieved from Microscopy Listserver Archives
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Thanks Paul,

Yes, this is what I told the person, I remember learning from Bendyan
that insulin is this way. ( and don't we wish that all antigens were
that strong!!)


I will talk to the researcher more, I suspect they may be asking because
their immuno didn't work as he was concerned about retrieval, but I'll
find out if he tried it anyway.

Lou Ann

Webster, Paul wrote:
} Dear Lou Ann,
}
} The rule that glutaraldehyde should not be used for immunolabeling is not
} absolute. Before attempting all sorts of different retrieval techniques why
} not give the immunolabeling a try. If the antigen is fairly abundant and not
} surrounded by lots of other proteins, you may find it works.
}
} Once you have a result from the simple immunolabeling experiment, and it
} shows not signal, then try complicating things by attempting antigen
} retrieval.
}
} There are lots of examples in the literature where high concentrations of
} glutaraldhyde were used for immunolabeling (and even Epo-embedded tissues).
}
} Paul.
}
}
}
}
} } From: {lamiller-at-uiuc.edu}
} } Reply-To: {lamiller-at-uiuc.edu}
} } Date: Thu, 12 Jul 2007 12:54:43 -0500
} } To: {pwebster-at-hei.org}
} } Subject: [Microscopy] Taking the gluteraldehyde out
} }
} }
} }
} }
} } ----------------------------------------------------------------------------
} } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
} } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
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} }
} } Dear Folks on the Microscopy List,
} }
} }
} } I have a researcher who wants to do immuno labeling, but the sample is
} } already in gluteraldehyde.
} }
} } I've always gone on the premise that gluteraldehyde is permanent, and
} } always suggest 4% parafromaldehyde or parafromaldehyde with the barest
} } littlest amount of gluteraldehyde ( {1%).
} }
} }
} } I am not familiar with any retrieval protocols, do you know of any for
} } gluteraldehyde, where the Ag site actually is affected by the
} } gluteraldehyde?
} }
} }
} }
} } Thanks,
} }
} }
} } 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
} }
} }
} } ==============================Original Headers==============================
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--
~~~~~~~~~~~~~~~~~~~~~~~~~~~~
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


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From: dkoleary-at-verizon.net
Date: Thu, 12 Jul 2007 13:50:07 -0500
Subject: [Microscopy] LM Workshop on Digital Photomicrography

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html

New York Microscopical Society
Bernard Friedman Memorial Workshop
Digital Photomicrography with the Light Microscope
August 21, 22 & 23, 2007

A hands-on workshop using professional microscopes and cameras designed for
photomicrography combined with the theory of their use.

This workshop teaches the fundamentals of taking digital micrographs through
transmitted light microscopes for illustrative purposes. Typical subjects
covered will be:

Getting the Microscope Ready for Image Recording
Centering of Objectives and Condenser Koehler Illumination
Significance of the Aperture Magnification and Resolution
Parfocality of Camera and Visualized Image

Digital Camera and Driver
Electronic image sensors Histograms Gamma
Settings
White Balance Background Subtraction Compression of Image
Files

Imaging Software used in the Course
Photoshop Elements:
ImageJ: Measurements of length and area, scale bar


Integration of Digital Images into Documents
Microsoft Word:
Importing images into MS Word


Instructors
Jan Hinsch, formerly of Leica Microsystems, Dr. Angela Klaus,
Seton Hall University
Mel Pollinger, Boston Scientific, Don O'Leary, NYMS .

The Workshop will be held at NYMS Headquarters at 30 N. Mountain Avenue,
Montclair, NJ from 9:00 AM to 5:00 PM. Lunch will be supplied
Cost will be $ 550 For Members $ 580 for nonmembers (includes membership)
Attendance is limited to the first 12 applicants.

For further information e-mail dkoleary-at-verizon.net or call (201) 368-8849

To register please return the form below with payment to: NYMS, C/o Donald
O'Leary, 10 Sampson Street. Unit 113, Saddle Brook, NJ 07663
_____________________________________________________
Digital Photomicrography with the Light Microscope
August 21, 22 & 23, 2007

Name_________________________________________________________________
Address_______________________________________________________________
City____________________________State_____________Zip_____________
Home Phone__________________________Wrk Phone____________________
e-Mail__________________________________________________________

Don O'Leary
Treasurer EAS 2007


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From: cammer-at-aecom.yu.edu
Date: Thu, 12 Jul 2007 16:22:27 -0500
Subject: [Microscopy] Zeiss Axioskop II uneven illumination

Contents Retrieved from Microscopy Listserver Archives
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We've been having problems for years getting flat illumination across
our images with a Zeiss AxioSkopII with a Zeiss Axiocam.

The unevenness is not noticeable when we have high contrast
histological stains, but with very low contrast samples, the corners
of the images are noticeably dark.

We've identified one source of the unevenness that causes problems
specifically at the right edges of our images. The ND filters in the
illumination light path reflect off each other (examples at
http://www.aecom.yu.edu/aif/temp/ZeissAxioSkopII/ ). If we take out
the ND filters, the field is far more even; however, the light is too
bright and turning down the voltage to the halogen lamp throws off
the color temperature.

Also, the unevenness is not gone ocmpletely.

Has anybody else noticed this problem and, if so, how have you solved it?

Thanks.

-Michael
____________________________________________________________________________
Michael Cammer Analytical Imaging Facility Albert Einstein Coll. of Med.
URL: http://www.aecom.yu.edu/aif/


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From: jmkrupp-at-ucsc.edu
Date: Thu, 12 Jul 2007 16:57:31 -0500
Subject: [Microscopy] Funding equipment

Contents Retrieved from Microscopy Listserver Archives
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Greetings

A few weeks ago I posed a question about how to take a look at some
bones for a quasi-CSI type project. I received several kind and
thoughtful replies, including some that suggested using an
environmental SEM.

Well, duh, what a good idea. Problem is, I don't have an ESEM and am
at a loss as to how to get one. Thus today's question.

How are new instruments funded? I have always assumed that some big
time PI would get grants and new instruments would just come rolling
in. Maybe this works if you have high rollers, and maybe it works for
their own labs, but what about a low rolling central campus facility
that does a little of this and a little of that for everyone?

We really don't have the critical mass needed to leverage much of an
extramural grant and our instruments are all approaching 25 years
old. They still work, but are not modern or up to date by any means.

If I ask for $$ from the Dean, the answer is usually 'Wow! I didn't
know they cost so much. Can you get by with what you have? There is
an emergency in blah blah's lab and we are out of money for this
year.' Kind of a management by crisis strategy. Is it like this
everywhere? Does anyone really have any long term infrastructural
planning in place?

So, does anyone know if there are actual institutions that plan for
replacing old equipment with institutional funds or are all
microscopes funded by extramural grants? I know about cost sharing,
but without the significant projects and faculty interest, we never
get that far. Sometimes I think I know how that slide rule salesman
must have felt when HP calculators hit the market.

Today I am faced with the prospect of spending $10K or so if I can
get the funding from the Dean to repair a 25 year old SEM or replace
it with a 'new', only 15 year old, used SEM I picked up as a donation
for about the same cost. Neither one would be an ESEM, and neither
one could be considered up to date.

What's a body to do? Enquiring minds want to know!

Thanks

Jon
--

Jonathan Krupp
Microscopy & Imaging Lab
C230 Earth & Marine Science
University of California
Santa Cruz, CA 95064
(831) 459-2477
jmkrupp-at-ucsc.edu

I rode in AIDS/Lifecycle 6 SF to LA June 3 - 9, 2007 to raise money
for the San Francisco AIDS Foundation. Visit
http://www.aidslifecycle.org for more information about the ride or
http://www.aidslifecycle.org/5482 to make a donation.

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From: anita.garg-at-grc.nasa.gov
Date: Thu, 12 Jul 2007 21:55:29 -0500
Subject: [Microscopy] viaWWW: Veleta: TEM CCD camera??

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Email: anita.garg-at-grc.nasa.gov
Name: Anita Garg

Organization: NASA GRC

Title-Subject: [Filtered] Veleta: TEM CCD camera??

Question: We are planning to buy a side mounted CCD camera for our CM200 TEM. We are considering the "2k X 2k Veleta" from Olympus SIS. We would like to hear the comments from the experienced users. Any suggestions regarding the "Diffraction Software" would be highly appreciated too.
TIA
Anita

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From: cvierret-at-umr.edu
Date: Thu, 12 Jul 2007 21:56:09 -0500
Subject: [Microscopy] viaWWW: Measuring Beam Diameter

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Email: cvierret-at-umr.edu
Name: Clarissa Wisner

Organization: University of Missouri At Rolla, for now

Title-Subject: [Filtered] Measuring Beam Diameter

Question: Hello,

I have been charged with measuring the beam diameter in our scopes. I have researched some methods and most involve a Si knife. Do any of you have a method that you have used to measure beam diameter and if so where did you obtain your materials. Any and all information would be appreciated.

Thanks

Clarissa Wisner
SEM Specialist
G6 MRC
University of Missouri
Rolla, Mo 65409
cvierret-at-umr.edu
573-341-4393



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From: nizets2-at-yahoo.com
Date: Fri, 13 Jul 2007 08:42:02 -0500
Subject: [Microscopy] Taking the gluteraldehyde out

Contents Retrieved from Microscopy Listserver Archives
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Hi All

First of all, as a neutral using all sorts of SEM from all the
manufacturers, if you have very little money or operating time to spend with
the instrument do not get hung up on an ESEM.

There are a mass of simple techniques that allow us to look at even the most
difficult specimens on an "ordinary" SEM. A conventional instrument is far
more likely to be available than a dedicated ESEM and most well known
models, even 15 to 20 years old, will serve you very well.

I have just run a course on a 20 year old Camscan in its new (second hand)
home, complete with EDS, it is more than capable of imaging at 30,000X.

Good luck

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: {jmkrupp-at-ucsc.edu}
To: {protrain-at-emcourses.com}
Sent: Thursday, July 12, 2007 11:00 PM

Hi Lou Ann,

The chances are weak but it is definitely worth a try.
However one important problem with glutar fixation is
that it gives autofluorescence, so use another
detection method. It is claimed that NaBH4 can block
the autofluorescence but it didn't really work with me
(probably depends on the intensity and pattern of the
specific labeling).

Stephane


--- lamiller-at-uiuc.edu wrote:

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}
} Thanks Paul,
}
} Yes, this is what I told the person, I remember
} learning from Bendyan
} that insulin is this way. ( and don't we wish that
} all antigens were
} that strong!!)
}
}
} I will talk to the researcher more, I suspect they
} may be asking because
} their immuno didn't work as he was concerned about
} retrieval, but I'll
} find out if he tried it anyway.
}
} Lou Ann
}
} Webster, Paul wrote:
} } Dear Lou Ann,
} }
} } The rule that glutaraldehyde should not be used
} for immunolabeling is not
} } absolute. Before attempting all sorts of different
} retrieval techniques why
} } not give the immunolabeling a try. If the antigen
} is fairly abundant and not
} } surrounded by lots of other proteins, you may find
} it works.
} }
} } Once you have a result from the simple
} immunolabeling experiment, and it
} } shows not signal, then try complicating things by
} attempting antigen
} } retrieval.
} }
} } There are lots of examples in the literature where
} high concentrations of
} } glutaraldhyde were used for immunolabeling (and
} even Epo-embedded tissues).
} }
} } Paul.
} }
} }
} }
} }
} } } From: {lamiller-at-uiuc.edu}
} } } Reply-To: {lamiller-at-uiuc.edu}
} } } Date: Thu, 12 Jul 2007 12:54:43 -0500
} } } To: {pwebster-at-hei.org}
} } } Subject: [Microscopy] Taking the gluteraldehyde
} out
} } }
} } }
} } }
} } }
} } }
}
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} } }
} } } Dear Folks on the Microscopy List,
} } }
} } }
} } } I have a researcher who wants to do immuno
} labeling, but the sample is
} } } already in gluteraldehyde.
} } }
} } } I've always gone on the premise that
} gluteraldehyde is permanent, and
} } } always suggest 4% parafromaldehyde or
} parafromaldehyde with the barest
} } } littlest amount of gluteraldehyde ( {1%).
} } }
} } }
} } } I am not familiar with any retrieval protocols,
} do you know of any for
} } } gluteraldehyde, where the Ag site actually is
} affected by the
} } } gluteraldehyde?
} } }
} } }
} } }
} } } Thanks,
} } }
} } }
} } } 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
} } }
} } }
} } } ==============================Original
} Headers==============================
} } } 14, 17 -- From lamiller-at-uiuc.edu Thu Jul 12
} 12:43:08 2007
} } } 14, 17 -- Received: from expredir4.cites.uiuc.edu
} (expredir4.cites.uiuc.edu
} } } [128.174.5.187])
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} (8.12.11.20060308/8.12.8) with ESMTP id
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} 12 Jul 2007 12:43:08 -0500
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} (redkite.cvm.uiuc.edu
} } } [130.126.18.157])
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} (8.14.1/8.14.1) with ESMTP id
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} 12 Jul 2007 12:43:08 -0500
} } } (CDT)
} } } 14, 17 -- Message-ID: {4696682D.20101-at-uiuc.edu}
} } } 14, 17 -- Date: Thu, 12 Jul 2007 12:43:09 -0500
} } } 14, 17 -- From: Lou Ann Miller
} {lamiller-at-uiuc.edu}
} } } 14, 17 -- Reply-To: lamiller-at-uiuc.edu
} } } 14, 17 -- User-Agent: Thunderbird 2.0.0.4
} (Macintosh/20070604)
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} } } 14, 17 -- Subject: Taking the gluteraldehyde out
} } } 14, 17 -- Content-Type: text/plain;
} charset=ISO-8859-1; format=flowed
} } } 14, 17 -- Content-Transfer-Encoding: 7bit
} } } ==============================End of -
} Headers==============================
} } }
} }
} }
} }
}
} --
} ~~~~~~~~~~~~~~~~~~~~~~~~~~~~
} Lou Ann Miller, MT(ASCP)
} Service Supervisor
} Center for Microscopic Imaging
} College of Veterinary Medicine
} Rm 1204 VMBSB
} 2001 S Lincoln Ave
} Urbana, IL 61802
}
} 217/244-1567
} http://treefrog.cvm.uiuc.edu
}
}
} ==============================Original
} Headers==============================
} 9, 19 -- From lamiller-at-uiuc.edu Thu Jul 12 13:04:40
} 2007
} 9, 19 -- Received: from expredir4.cites.uiuc.edu
} (expredir4.cites.uiuc.edu [128.174.5.187])
} 9, 19 -- by ns.microscopy.com
} (8.12.11.20060308/8.12.8) with ESMTP id
} l6CI4djI008640
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} Jul 2007 13:04:40 -0500
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} 9, 19 -- Thu, 12 Jul 2007 13:04:39 -0500 (CDT)
} 9, 19 -- Message-ID: {46966D38.1090401-at-uiuc.edu}
} 9, 19 -- Date: Thu, 12 Jul 2007 13:04:40 -0500
} 9, 19 -- From: Lou Ann Miller {lamiller-at-uiuc.edu}
} 9, 19 -- Reply-To: lamiller-at-uiuc.edu
} 9, 19 -- User-Agent: Thunderbird 2.0.0.4
} (Macintosh/20070604)
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} 9, 19 -- To: "Webster, Paul" {PWebster-at-hei.org} ,
} microscopy-at-microscopy.com
} 9, 19 -- Subject: Re: [Microscopy] Taking the
} gluteraldehyde out
}
=== message truncated ===




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From: lcgould-at-med.cornell.edu
Date: Fri, 13 Jul 2007 08:43:25 -0500
Subject: [Microscopy] Re: Funding equipment

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html

Dear Jon-
It is an issue for all of us. I have a grant pending at the NIH for
a new TEM to replace my "baby", which was installed in 1982. Even
with really good projects, I'm on pins-and-needles waiting for our
score. The only other ways I've heard for people to get major
instruments (EMs, confocals, etc) without submitting shared
instrumentation grant applications were pooling of funds by a group
of investigators who then share the instrument exclusively, or being
fortunate enough to have generous private donations to your
institution earmarked for a specific piece of equipment. For the
latter, you need a really strong fund-raising group and/or a core
group of generous people of means seeking meaningful ways to get tax
deductions ;-).
In the meantime, try asking around at other institutions in your
region. Someone may have an ESEM with time available. Our local
orthopedic hospital (which has a research department) has one.
I will be interesting to read any other advice you receive about
alternate funding sources.
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

voice (212)746-6146
fax (212)746-8175
http://www.cornellcelldevbiology.org
http://www.cornellbiochem.org

==============================Original Headers==============================
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From: nizets2-at-yahoo.com
Date: Fri, 13 Jul 2007 08:48:53 -0500
Subject: [Microscopy] Re: viaWWW: Veleta: TEM CCD camera??

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html

I use the Megaview III from SIS with most
satisfaction.
The support I received from this company is not common
(in terms of speed and efficiency) and it is a
pleasure for me to promote it if I can.
No other interest in this company other than to make
other customers happy ;-)

Stephane


--- anita.garg-at-grc.nasa.gov wrote:

}
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} Email: anita.garg-at-grc.nasa.gov
} Name: Anita Garg
}
} Organization: NASA GRC
}
} Title-Subject: [Filtered] Veleta: TEM CCD camera??
}
} Question: We are planning to buy a side mounted CCD
} camera for our CM200 TEM. We are considering the "2k
} X 2k Veleta" from Olympus SIS. We would like to hear
} the comments from the experienced users. Any
} suggestions regarding the "Diffraction Software"
} would be highly appreciated too.
} TIA
} Anita
}
}
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} ==============================Original
} Headers==============================
} 6, 11 -- From zaluzec-at-microscopy.com Thu Jul 12
} 21:55:29 2007
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} 6, 11 -- Subject: viaWWW: Veleta: TEM CCD camera??
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}



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==============================Original Headers==============================
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From: bingber-at-srrc.ars.usda.gov
Date: Fri, 13 Jul 2007 09:53:19 -0500
Subject: [Microscopy] Information on Confocal Microscopy Systems

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html

I need information on confocal systems-pricing, accessories, etc. Vendors may contact me offline at my gov address. Users may contact me either off or online. Thank you for your assistance.

Bruce F. Ingber, Biologist
Electron Microscopy
USDA-ARS, SRRC
CSQ-EM
1100 Robert E. Lee Blvd.
New Orleans, LA 70124
bingber-at-srrc.ars.usda.gov
bingber46-at-hotmail.com
504-286-4270 desk phone
504-782-6323 cell



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From: cammer-at-aecom.yu.edu
Date: Fri, 13 Jul 2007 12:18:12 -0500
Subject: [Microscopy] Re: Zeiss Axioskop II uneven illumination

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html

Thank you for the software suggestions (also Image Arithmetic).

I think the biggest problems are the reflections and an aperture that
doesn't match the rest of the optics. Perhaps it's a simple parts
mismatch. I doubt I'm being too picky expecting a high end research
microscope to have even illumination edge-to-edge.

-Michael

At 11:34 AM 07/13/07, you wrote:
} I thought I would mention it in case you weren't familiar with it. In
} the Axiovison software (V4.?) there is a software button you can enable
} called "shading correction"
} It appears to even out the image illumination.
} I have heard rumors in the lab that it is sometimes used instead of
} properly aligning the microscope: )

____________________________________________________________________________
Michael Cammer Analytical Imaging Facility Albert Einstein Coll. of Med.
URL: http://www.aecom.yu.edu/aif/


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From: phillipst-at-missouri.edu
Date: Fri, 13 Jul 2007 15:15:08 -0500
Subject: [Microscopy] paraffin dewaxing

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html

I work mostly with plastic resin embedded tissue and only sporadically with
paraffin. Is there a rule of thumb for how many slides can be dewaxed per
volume before it is necessary to switch solutions. I understand the concept
of moving Xylene bath 3 to bath 2 and bath 2 to bath 1 and fresh into bath
3 but at what point? Thanks, Tom



Thomas E. Phillips, PhD
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400

573-882-4712 (office)
573-882-0123 (fax)
PhillipsT-at-missouri.edu
http://www.biology.missouri.edu/faculty/phillips.html
http://www.biotech.missouri.edu/mcc/


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From: rcmoretz-at-gmail.com
Date: Fri, 13 Jul 2007 15:46:36 -0500
Subject: [Microscopy] Re: paraffin dewaxing

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html

Tom:

On the automatic Leica stainers used (and in the manual procedures set
up in our SOPs) the magic number was 400 slides, volume of each
station approx 450ml, 3x xylene, 2x 100% EtOH, 2x 95% EtOH. Hope that
helps.

Roger Moretz, Ph.D.
Principally from nowhere, retired from the grind as of July 4th but
not dead yet.

On 7/13/07, phillipst-at-missouri.edu {phillipst-at-missouri.edu} wrote:
}
}
}
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} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
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} I work mostly with plastic resin embedded tissue and only sporadically with
} paraffin. Is there a rule of thumb for how many slides can be dewaxed per
} volume before it is necessary to switch solutions. I understand the concept
} of moving Xylene bath 3 to bath 2 and bath 2 to bath 1 and fresh into bath
} 3 but at what point? Thanks, Tom
}
}
}
} Thomas E. Phillips, PhD
} Professor of Biological Sciences
} Director, Molecular Cytology Core
} 2 Tucker Hall
} University of Missouri
} Columbia, MO 65211-7400
}
} 573-882-4712 (office)
} 573-882-0123 (fax)
} PhillipsT-at-missouri.edu
} http://www.biology.missouri.edu/faculty/phillips.html
} http://www.biotech.missouri.edu/mcc/
}
}
} ==============================Original Headers==============================
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From: Sven.Terclavers-at-med.kuleuven.be
Date: Fri, 13 Jul 2007 16:07:20 -0500
Subject: [Microscopy] paraffin dewaxing

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html

Hi Thomas,

In our lab, we change the solutions every 300, maximum 400 slides (500 ml)
or once per week, depending on which limit is reached first.

Sven Terclavers

-----Original Message-----
X-from: phillipst-at-missouri.edu [mailto:phillipst-at-missouri.edu]
Sent: Friday, July 13, 2007 10:18 PM
To: sven.terclavers-at-med.kuleuven.be

I work mostly with plastic resin embedded tissue and only sporadically with
paraffin. Is there a rule of thumb for how many slides can be dewaxed per
volume before it is necessary to switch solutions. I understand the concept
of moving Xylene bath 3 to bath 2 and bath 2 to bath 1 and fresh into bath
3 but at what point? Thanks, Tom



Thomas E. Phillips, PhD
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400

573-882-4712 (office)
573-882-0123 (fax)
PhillipsT-at-missouri.edu
http://www.biology.missouri.edu/faculty/phillips.html
http://www.biotech.missouri.edu/mcc/


==============================Original Headers==============================
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From: levilr-at-hughes.net
Date: Sat, 14 Jul 2007 18:07:43 -0500
Subject: [Microscopy] Manuals for ISI/Topcon SX30

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html

I have an ISI/Topcon SX30 microscope with no manuals/schematics. If anyone
has a set of manuals/schematics as pdfs or as paper versions I'd appreciate
a copy. Please contact me.

Thank you

Lee Levine
Process Solutions Consulting
8009 George Road,
New Tripoli, PA 18066
mobile 610-248-2002, fax 610-285-4575
email levilr-at-hughes.net


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From: tivol-at-caltech.edu
Date: Sat, 14 Jul 2007 18:41:19 -0500
Subject: [Microscopy] Re: viaWWW: Measuring Beam Diameter

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On Jul 12, 2007, at 7:57 PM, cvierret-at-umr.edu wrote:

} I have been charged with measuring the beam diameter in our scopes. I
} have researched some methods and most involve a Si knife. Do any of
} you have a method that you have used to measure beam diameter and if
} so where did you obtain your materials. Any and all information would
} be appreciated.
}
Dear Clarissa,
When I did this, I exposed a piece of film at a high enough mag that
the maximum intensity did not saturate the film, and the beam still fit
onto the film. I then scanned the film and got a measure of intensity
vs. distance from the beam center. I was interested in determining the
minimum probe size I could get, which happened at the highest Wehnelt
bias and first condenser lens settings, so the intensity was low enough
to do this. You could try the same thing with a CCD by first going to
a high enough mag so that the CCD was not saturated, then taking other
images at lower mags, if necessary, to include all the beam. You might
have to use the beam stop to intercept the brightest part of the beam
at these lower mags.
Yours,
Bill Tivol, PhD
EM Scientist
Electron Cryo-Microscopy Facility
Broad Center, Mail Code 114-96
California Institute of Technology
Pasadena CA 91125
(626) 395-8833
tivol-at-caltech.edu


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From: zaluzec-at-aaem.amc.anl.gov
Date: Sun, 15 Jul 2007 17:17:37 -0500
Subject: [Microscopy] Administrivia: Spam Message

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html

Colleagues

A bit of SPAM Email got through the filters today.

I am investigating where the failure in my filter software
is, so please ignore a possible forged Email from microscopytoday.

Nestor
Your Friendly Neighborhood SysOp

--

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5, 14 -- From: "Nestor J. Zaluzec" {zaluzec-at-aaem.amc.anl.gov}
5, 14 -- Subject: Administrivia: Spam Message
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From: zaluzec-at-microscopy.com
Date: Sun, 15 Jul 2007 19:13:45 -0500
Subject: [Microscopy] Administrivia: Spam Update

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html

Colleagues

I believe I have found the loop hole and plugged it.

In case your interested the forged Email
originated from a site in Russia, who forged a rr.com
Email address from Microscopy-Today headers.


Nestor


==============================Original Headers==============================
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From: mike-at-bitplane.com
Date: Mon, 16 Jul 2007 10:31:00 -0500
Subject: [Microscopy] Positions Available in the USA

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html

Bitplane is expanding its US team and asks for applications for the
following two positions. Feel free to forward this e-mail to anyone who
might be interested.

Position #1 - Central and South Eastern Region Sales Representative US /
Canada

Bitplane is looking for a biologist with strong computer skills and at least
one year of hands-on experience using a confocal or similar 3D advanced
light microscope.
The duties of this position include:

. Customer visits and analysis of customer's imaging needs.
. Demonstration of the software and onsite work with the customer
. Organization of exhibitions and workshops
. Sales support of existing customers

The candidate is expected to have an outgoing personality with strong
communication skills and should look forward to increased responsibility. At
least 50% travel will be required. Representative is required to live in the
territory they cover. Benefits include a base salary, performance based
commission, 401K plan, healthcare, and vacation. Representative will work
out of a home office. We offer a team of 18 people, fun to work with, and a
truly international environment that provides the resources required to
grow. We don't mind if the candidate does not have much business experience
and we are prepared to show him/her the sales skills at the job.

Position #2 - Sales Development Coordinator

Bitplane is looking for a candidate with a science background and knowledge
of the microscope community. The duties of this position include

. Identifying potential new regional customers via web searches, literature
searches, marketing campaigns, trade shows, and customer referrals.
. Introduction of Bitplane products and services to potential customers via
email, phone, and Webex.
. Understanding potential customers needs related to products offered by
Bitplane.
. Organizing workshops and demonstrations for regional sales representatives


The candidate is expected to have an outgoing personality, is detail
oriented, articulates clearly, exhibits a high level of professionalism, and
exceptional organization.Travel is not required. Representative is/will be
based in the Minneapolis / Saint Paul area of MN. Benefits include a base
salary, 401K plan, healthcare, and vacation. Representatives will work out
of a home office and travel is not required on routine basis. We offer a
team of 18 people, fun to work with, and a truly international environment
that provides the resources required to grow.

Applicants should reply with a letter stating what position they are
interested in, why they are qualified for the position, and enclose their
CV.

With best regards,
Mike


Bitplane Inc.
Michael C. Wussow
Vice President and General Manager Bitplane Inc.

Cell Phone: 651-336-4600
Fax: 866-691-9112
Toll Free: 1-888-3D-BITPX (332-4879)
Visit Our Web Site At: www.bitplane.com





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From: fittonro-at-luther.edu
Date: Mon, 16 Jul 2007 14:15:50 -0500
Subject: [Microscopy] Gatan 791 manual

Contents Retrieved from Microscopy Listserver Archives
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Greetings,

I could use either a paper or a pdf copy of the manual for a Gatan
791 CCD camera for a TEM that includes installation instructions.
Or, specific advice on the air line connection and pressure for a
JEOL 1200.

Thanks in advance!

Robert


Robert Fitton
Teaching Associate/Director of Labs
Luther College
Department of Biology
700 College Drive
Decorah, IA 52101
563-387-1559 Voice
563-387-1080 FAX
fittonro-at-luther.edu





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From: tina-at-pbrc.hawaii.edu
Date: Mon, 16 Jul 2007 19:41:40 -0500
Subject: [Microscopy] pipeting resin components

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html

Hi, all-

Here's kind of a dumb question. How do you all measure out embedding resin
catalyst? I've only been doing it for 31 years... I used to use tuberculin
syringes - had to hurry because they sort of melt. The current crop of
facility users seem dumbfounded that I would use anything but a
pipetter. But I have now had another new pipetter get gummed up with some
resin component, probably catalyst. Does anyone have a favorite method???

Aloha with exasperation,
Tina

****************************************************************************
* Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu *
* Biological Electron Microscope Facility * (808) 956-6251 *
* University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf*
****************************************************************************


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From: dsherman-at-purdue.edu
Date: Mon, 16 Jul 2007 20:03:20 -0500
Subject: [Microscopy] Re: pipeting resin components

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html

Tina,

Ah yes, I remember those days of plugged pipetters!

To eliminate this we weigh all resin components into a dispo beaker. We can
get close enough with a decent 3 decimal place scale to get consistent
results and use only disposable pipettes for all components.

Our scale sits in a hood so is protected by a draft shield. This has gotten
rather messy over the years as students occasionally use acetone on the
polyethylene shield to clean up spills........but still works. A piece of
aluminum foil on the pan has worked to prevent resin from getting onto scale
components that would affect function.

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



On 7/16/07 8:45 PM, "tina-at-pbrc.hawaii.edu" {tina-at-pbrc.hawaii.edu} wrote:

}
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}
} Hi, all-
}
} Here's kind of a dumb question. How do you all measure out embedding resin
} catalyst? I've only been doing it for 31 years... I used to use tuberculin
} syringes - had to hurry because they sort of melt. The current crop of
} facility users seem dumbfounded that I would use anything but a
} pipetter. But I have now had another new pipetter get gummed up with some
} resin component, probably catalyst. Does anyone have a favorite method???
}
} Aloha with exasperation,
} Tina
}
} ****************************************************************************
} * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu *
} * Biological Electron Microscope Facility * (808) 956-6251 *
} * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf*
} ****************************************************************************
}
}
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From: shah0211-at-umn.edu
Date: Mon, 16 Jul 2007 21:00:20 -0500
Subject: [Microscopy] viaWWW: Diamond knife

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Email: shah0211-at-umn.edu
Name: Neha Shah

Organization: U of Minnesota

Title-Subject: [Filtered] Diamond knife

Question: Our lab is looking to buy a diamond knife and we are debating between PELCO and DiaTome. Our EM facility currently uses DiaTome and it works well. Does anyone have information about PELCO diamond knife and whether they work well with biological samples?



---------------------------------------------------------------------------

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From: nizets2-at-yahoo.com
Date: Mon, 16 Jul 2007 23:51:13 -0500
Subject: [Microscopy] pipeting resin components

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html

+1

Stephane

} Tina,
}
} Ah yes, I remember those days of plugged pipetters!
}
} To eliminate this we weigh all resin components into
} a dispo beaker. We can
} get close enough with a decent 3 decimal place scale
} to get consistent
} results and use only disposable pipettes for all
} components.




____________________________________________________________________________________
Be a better Heartthrob. Get better relationship answers from someone who knows. Yahoo! Answers - Check it out.
http://answers.yahoo.com/dir/?link=list&sid=396545433

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From: oshel1pe-at-cmich.edu
Date: Tue, 17 Jul 2007 07:44:16 -0500
Subject: [Microscopy] Re: pipeting resin components

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html

Retry -- the spam filter rejected me.

Glass transfer pipettes and rubber bulbs. Measure by weight.

Phil

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From: nizets2-at-yahoo.com
Date: Tue, 17 Jul 2007 08:07:41 -0500
Subject: [Microscopy] microcalorimetric EDS

Contents Retrieved from Microscopy Listserver Archives
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Hi!

I wondered where is the development of this technique.
It obviously uses supraconductivity, so it needs
temperatures lower than LN2, how is this temperature
maintained?
Is anyone in the field, or have had the chance to use
this technology?

Best regards,

Stephane



____________________________________________________________________________________
TV dinner still cooling?
Check out "Tonight's Picks" on Yahoo! TV.
http://tv.yahoo.com/

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From: lcgould-at-med.cornell.edu
Date: Tue, 17 Jul 2007 08:18:59 -0500
Subject: [Microscopy] Re: pipeting resin components

Contents Retrieved from Microscopy Listserver Archives
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Aloha, Tina-
I use a pipettor, but I use the big ones (1000microliters) to
measure out 300-600 microliters of catalyst so that I don't risk
pulling the gooey stuff up into the works. If I need to make a large
volume, I measure out the catalyst in 2 shots. Simple, but fairly
effective. I haven't had a gummed-up pipettor in a while. (Now
watch, it will happen this week!)
Lee
--
Leona Cohen-Gould, M.S.
Sr. Staff Associate
Director, Electron Microscopy & Histology Core Facility
Manager, Optical Microscopy Core Facility
Weill Cornell Medical College

voice (212)746-6146
fax (212)746-8175
http://www.cornellcelldevbiology.org
http://www.cornellbiochem.org

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From: fli-at-estee.com
Date: Tue, 17 Jul 2007 08:30:11 -0500
Subject: [Microscopy] AskAMicroscopist: FEI Phenom Electron-light microscope does it

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This Question was submitted to Ask-A-Microscopist by (fli-at-estee.com)
from http://www.microscopy.org/Ask-A-Microscopist/Ask-A-Microscopist.html on Tuesday, July 17, 2007 at 08:08:29
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Email: fli-at-estee.com
Name: Foo Li

Organization: Estee Lauder

Education: Graduate College

Location: Melville, NY 11747

Title: FEI Phenom Electron-light microscope

Question: Is the Phenom the real deal? 30 nm resolution is not all that bad especially when dealing with emulsions. I need an opinion of this product from actual Phenom users out there.

---------------------------------------------------------------------------

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From: dac-at-research.umass.edu
Date: Tue, 17 Jul 2007 09:12:08 -0500
Subject: [Microscopy] pipeting resin components

Contents Retrieved from Microscopy Listserver Archives
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From the kitchen of "Cowboy Cooking"....

Trying to weigh the catalyst in the fume hood, I have a problem of air
flow of the hood disturbing the balance reading. I once held my nose
(away from the hood) and measured the number of drops of catalyst from a
particular smallish PE disposable pipet required to make the desired
weight. It is very repeatable if you let the drops form (don't squirt)
and use the same style pipet. I often mix resin batches in the 50cc
range that require 0.2g of catalyst. I weigh the main components out in
the lab (scissor off the tip of larger dispo plastic pipet for the
viscous resins), mix well, then move to the hood and add the drops of
catalyst. For my pipets and DMP-30 or DMAE I use 12 drops for 0.2g.

Since Spurr's resin works fine with half the normal catalyst for "Long
Pot Life" mix, the tiny variations in the drop method certainly will
have very little effect on the final product.

Cheers,

Dale

lcgould-at-med.cornell.edu wrote:
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} Aloha, Tina-
} I use a pipettor, but I use the big ones (1000microliters) to
} measure out 300-600 microliters of catalyst so that I don't risk
} pulling the gooey stuff up into the works. If I need to make a large
} volume, I measure out the catalyst in 2 shots. Simple, but fairly
} effective. I haven't had a gummed-up pipettor in a while. (Now
} watch, it will happen this week!)
} Lee

==============================Original Headers==============================
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From: nicholas.ritchie-at-nist.gov
Date: Tue, 17 Jul 2007 09:27:26 -0500
Subject: [Microscopy] Re: Microcalorimetric EDS

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html

Stephane,
We've been working with one of the earliest microcalorimeters for
about 5 years. They are beautiful spectrometers if you are willing to
work around their shortcomings. Ours is capable of collecting energy
dispersive spectra over energy ranges between a few hundred eV and 8 keV
with resolutions of around 6 eV. Count rates are low - around one to
two hundred counts per second.
The energy dispersion is achieved by measuring the temperature
increase in a block of normal (non-superconducting) metal. The
thermometer is a superconductor held right at the temperature at which
it makes the transition from normal conduction to superconduction. The
change in resistance with change in temperature is large and can be read
out using SQUIDS (superconducting quantum interference devices).
The device is maintained at liquid He temperatures (~4 degrees) all
the time and cooled to ~100 millikelvin using an adiabatic
demagnetization refrigerator for operation. It can remain at 100
millikelvin for about 10 hours.
To the best of my knowledge there are not currently any vendors
selling microcalorimeters although there were in the past. There is at
least one commercial unit in development. The cost is likely to be in
the hundreds of thousands of dollar range per unit.
Microcalorimeters are nothing like Si(Li) or SDD in terms of ease of
use. Ours takes a couple of hours to cycle down to temperature and to
lock the detector onto the edge of the superconducting transition. They
require constant fiddling with half-a-dozen or more amplifiers and field
coils. Someday they may be refined enough for routine analysis but
today they remain laboratory experiments.

Sincerely,

Nicholas Ritchie


nizets2-at-yahoo.com wrote:
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Hi!

I wondered where is the development of this technique.
It obviously uses supraconductivity, so it needs
temperatures lower than LN2, how is this temperature
maintained?
Is anyone in the field, or have had the chance to use
this technology?

Best regards,

Stephane



____________________________________________________________________________________

TV dinner still cooling? Check out "Tonight's Picks" on Yahoo! TV.
http://tv.yahoo.com/

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From: smentkow-at-crd.ge.com
Date: Tue, 17 Jul 2007 11:35:36 -0500
Subject: [Microscopy] new short course at MM2007 - space still available

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A new short course "X06-Surface Analysis" is being offered at this years
MM meeting in Ft. Lauderdale Florida (Sun, Aug 5, 2007) at 9am in room
222 of the convention center. We have been informed that space is still
available. This is an excellent opportunity to learning more about state
of the art surface analysis instrumentation through examples.

Course summary:
For many biological and materials systems, the properties of the
specimen's outer surface dictate its performance. Surface analysts are
being asked to detect specimen components present in ever lower
concentrations and within smaller spatial dimensions. This short course
will describe and contrast the main surface analytical instruments:
Auger Electron Spectroscopy (AES), X-ray Photoelectron Spectroscopy
(XPS), Secondary Ion Mass Spectrometry (SIMS), and Scanning Probe
Microscopy (SPM). Emphasis will be placed on the principles of each
technique and on data interpretation. The limitations of each technique
will be discussed. Upon completion of this short course, the attendee
will be able to properly select which state-of-the-art surface analysis
technique(s) should be used to address spectroscopy, imaging, and depth
profiling analysis requests from their colleagues/customers.

Vincent S. Smentkowski
GE Global Research
1 Research Circle
Niskayuna, NY 12309 USA
General Electric Company
T 518 387 5467
F 518 387 6972
E smentkow-at-crd.ge.com




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From: jd-at-laddresearch.com
Date: Tue, 17 Jul 2007 13:08:17 -0500
Subject: [Microscopy] Re: viaWWW: Diamond knife

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html

The Pelco and Diatome are both high quality knives which should work
very well with biological samples.

Having said that, we feel the Drukker knife is the best value and
quality knife on the market. You might want to check out the UL3 for
the best in both quality and value.

http://www.laddresearch.com/New_Products/Drukker_Diamond_Knives/drukker_diamond_knives.html

John Arnott

Disclaimer: Ladd Research sells diamond knives and other EM supplies
and accessories

Ladd Research
83 Holly Court
Williston, VT 05495

On-line Catalog: www.laddresearch.com

Telephone: 1-802-658-4961 (anywhere)
Toll Free 1-800-451-3406 (US)
Fax: 1-802-660-8859

e-mail: sales-at-laddresearch.com

At 10:07 PM 7/16/2007, you wrote:



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From: tina-at-pbrc.hawaii.edu
Date: Tue, 17 Jul 2007 16:53:40 -0500
Subject: [Microscopy] Re: pipeting resin components

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html

I had to giggle at the many replies to my cry of frustration about
measuring catalyst for resin! Like Tamara, over the years I have become
adept at sequentially pouring the resin components into a tri-cornered
beaker, stirring magnetically, then adding catalyst. Many years ago I used
to measure out the components with syringes and then laboriously clean the
syringes and keep them for the next use. Once I switched to weighing I
kept using a tuberculin syringe to measure catalyst because my crappy
balance couldn't weigh small amounts and did not do well in the fume
hood; the draft shield was stolen years ago. Then I got mentally
stuck. My syringes are desperately old, and the black rubber seal has
been melting when in contact with the catalyst.

Now when I train people they look at me like I'm crazy when I pour, so I
let them transfer components to the beaker on the balance with disposable
plastic transfer pipets. I *do not allow* glass pipets! How many of you
have sectioned glass chips?

The current crop of students don't seem to know how to use graduated
pipets and two- or three-button bulbs, so I have been letting them use the
pipetters to measure catalyst. They all know how to use pipetters, so it
was an easy fix. In my cranky old age, though, I've been insisting they
learn other ways to measure! I don't know how resin got up into the
pipetters; I would have thought it was impossible, but it keeps happening.

I keep all wastes: transfer pipets, tips, syringes, and beakers, and
polymerize them, thinking it makes it all less toxic when it goes to the
landfill or waste-to-energy plant.

Tina's Tip: I put my molds and labels in the oven overnight to bake out,
and now I never get bubbles!

Aloha,
Tina

****************************************************************************
* Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu *
* Biological Electron Microscope Facility * (808) 956-6251 *
* University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf*
****************************************************************************


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From: PWebster-at-hei.org
Date: Tue, 17 Jul 2007 19:21:10 -0500
Subject: [Microscopy] Re: pipeting resin components

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html

Dear All,

It seems that in many ways my lab may be unique in its way of handling
resin. Many years ago I threw out the concept of solutions A and B , and the
WPE values and went instead to a method first proposed to me by Sus Ito at
Harvard.

The method has worked consistently on three different continents and on a
few EM course in different parts of the world. It consists of mixing three
resin ingredients (but not the catalyst) in a large volume, aliquoting the
mixture into small glass, screw-top vials and storing them, tightly closed,
at -20 degrees for as long as it takes to finish the batch.

As resin is needed, a vial containing from 4 to 6 ml of resin is removed
from the freezer and warmed in the oven. The actual amount of resin in the
vial can be determined by comparing the resin level with a calibrated (and
marked) black vial). It is important to know the amount of resin in the
vail because this will determine the amount of catalyst to use.

Difficult to embed specimens can be soaked in the resin without catalyst for
many days, or the resin can be mixed with catalyst and used immediately. The
advantages are:

1. no messy resin preparation needed for each experiment.
2. no wasted resin components.
3. less handling of toxic components.
4. idiot-proof system ensuring reproducibility.
5. consistent results - the first vial is the same as the last vial.
6. no difficult measuring of catalyst (or any of the other components).
7. cleaner than other methods.

Want to know the method? Here it is:

Epon Resin (simplified recipe for making large batches of premixed resin)

Mix the following in a large glass jar (see below):

NMA 110 mL
DDSA 130 mL
Eponate 12 230 mL

When the ingredients have been well mixed, the resin is poured into small
tubes in aliquots of 4 to 6 mL and stored frozen. For use, warm a tube and,
using a glass pasteur pipette, add four drops of BDMA per milliliter (or 2
drops of DMP-30 if preferred). Mix well, use immediately and discard unused
resin (polymerization is a good idea).

Preparing and storing resin this way will ensure that resin ingredients are
not wasted and all the tubes will have resin of the same consistency. The
mixture can be tested before use by polymerizing one tube of resin. All
subsequent tubes of resin will have the same polymerization and sectioning
qualities as this first tube.

Glass jar tip: A simple tip for measuring the resin components is to
pre-calibrate a glass jar. In a clean jar, pour in 110mL of water, mark the
meniscus. Add an extra 13mL of water and mark the new meniscus, and finally,
add 230mL of water and mark the meniscus. (For those in the USA, Smuckers
jars work well - UK, Robinson's orange marmalade - it just has to be clear
glass!)

Pour out all the water and dry the jar well. It is now ready to be used for
measuring out the resin components and mixing them. Mix the three components
well before pouring the resin into the small vials. As the volume is
estimated just prior to the resin being used, there is no need to get into
complicated pipetting methods, just pour carefully.

As for glass pipettes (and I don't mean to be confrontational with the
following comments), I have been using them for many years in my specimen
preparation protocols. In addition to distributing catalyst, they are very
useful for collecting scraped cells from culture dishes, and for
transferring solutions from around very small specimens.

I am aware that glass fragments are supposed to enter embedded specimen
blocks and damage diamond knives, but most people I know also trim their
specimen blocks using glass knives, which is a more reasonable source of
glass contamination.

In all the years I have been using glass pipettes I have never come across
glass fragments in embedded tissues or cell pellets. I have seen hairs, dust
and large chinks of keratin, but no glass. If glass were present in the
blocks, I am sure it would be obvious in the initial trimming and facing
off, which, incidentally we do with an old diamond knife.

I send this method out for all to try. If anyone is brave enough to try it,
let me know if it was useful - I promise that it will change your life,
especially is you are part of a multi-user facility.

Best regards,

Paul Webster.


Paul Webster, Ph.D
House Ear Institute
2100 West Third Street
Los Angeles, CA 90057
(213) 273 8026
pwebster-at-hei.org





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I had to giggle at the many replies to my cry of frustration about
measuring catalyst for resin! Like Tamara, over the years I have become
adept at sequentially pouring the resin components into a tri-cornered
beaker, stirring magnetically, then adding catalyst. Many years ago I used
to measure out the components with syringes and then laboriously clean the
syringes and keep them for the next use. Once I switched to weighing I
kept using a tuberculin syringe to measure catalyst because my crappy
balance couldn't weigh small amounts and did not do well in the fume
hood; the draft shield was stolen years ago. Then I got mentally
stuck. My syringes are desperately old, and the black rubber seal has
been melting when in contact with the catalyst.

Now when I train people they look at me like I'm crazy when I pour, so I
let them transfer components to the beaker on the balance with disposable
plastic transfer pipets. I *do not allow* glass pipets! How many of you
have sectioned glass chips?

The current crop of students don't seem to know how to use graduated
pipets and two- or three-button bulbs, so I have been letting them use the
pipetters to measure catalyst. They all know how to use pipetters, so it
was an easy fix. In my cranky old age, though, I've been insisting they
learn other ways to measure! I don't know how resin got up into the
pipetters; I would have thought it was impossible, but it keeps happening.

I keep all wastes: transfer pipets, tips, syringes, and beakers, and
polymerize them, thinking it makes it all less toxic when it goes to the
landfill or waste-to-energy plant.

Tina's Tip: I put my molds and labels in the oven overnight to bake out,
and now I never get bubbles!

Aloha,
Tina

****************************************************************************
* Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu *
* Biological Electron Microscope Facility * (808) 956-6251 *
* University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf*
****************************************************************************


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From: wpchan-at-u.washington.edu
Date: Tue, 17 Jul 2007 19:45:10 -0500
Subject: [Microscopy] gatan camera

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html

Hi

I encountered a strange problem with a Gatan 689 Slow-Scan CCD camera
mounted in the 35 mm port above the viewing chamber of a Philips CM100
TEM. When I insert the camera, I can only see a uniform grey image. The
histogram shows a single line in the middle. Changing the exposure time
or increasing the illumination via condenser 2 has no effect. Allowing
the shutter to be normally closed, or putting the shutter to open also has
no effect.

There seemed to be no mechanical problem for inserting or retracting the
camera because the beam was blocked and blank as it should be.

We are still using DigitalMicrograph 2.5 on an old quadra 840 with system
7.1. I have trashed the preferences and used another copy of the software
with no improvement. When the camera is out, I can see the raw image in
unprocessed view; basically the dark reference image. With high tension
off, I used to see the same image when I insert the camera. But now I
only see a grey image. I don't think there is anything wrong with the CCD
or scintillator because I can see an after-image of the image I should be
seeing when I retract the camera. This suggested that the CCD was
exposed and formed an image but somehow not transferred to the computer.

Any suggestion or insight to solve this problem will be much appreciated.

--
Pang (Wai Pang Chan, wpchan-at-u.washington.edu, PAB A087, 206-685-1519)
The Biology Imaging Facility (http://staff.washington.edu/wpchan/if/)

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From: walck-at-southbaytech.com
Date: Tue, 17 Jul 2007 22:54:37 -0500
Subject: [Microscopy] Microcalorimetric EDS

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html

I just wanted to add a little information from what I learned in a
Statistical Physics course about adiabatic demagnetization 32 years ago as
an undergraduate. I worked in a low temperature Physics lab for a summer
and I just thought that the application of this was just so cool. (Please,
don't pardon the pun. It was intended.) Anyway, the original question
asked how it was maintained so I thought that I would add this little
description.

The adiabatic demagnetization process uses a paramagnetic salt. When the
high magnetic field is turned on, the magnetic moments of the salt aligns
with the field. In the Statistical Physics course there was a description
of the population of aligned moments over a "kT" term in an exponetional
term. The salt is thermally insulated well both mechanically and
radiation-wise by the liquid He cooled radiation shield. When the magnetic
field is suddenly switched, the population of moments is inverted. The
statistical system requires energy to switch the moments and align them with
the new magnetic field direction. Since the system is insulated, the only
place to take the heat from is from the phonon vibrations of the material
itself. The switching of the population into the aligned condition cools the
sample by taking the heat from the salt itself. You keep switching the
field and it will asymptotically reach a very low equilibrium temperature in
the milli-Kelvin range.

Cool, Isn't it. I know, I know. I'm a nerd.


-Scott

Scott D. Walck, Ph.D.
Technical Director
South Bay Technology, Inc.
1120 Via Callejon
San Clemente, CA 92673

US Toll Free: 1-800-728-2233
Tel: (949) 492-2600
Fax: (949) 492-1499

www.southbaytech.com
walck-at-southbaytech.com



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From: kenner.rita-at-marshfieldclinic.org
Date: Wed, 18 Jul 2007 00:37:52 -0500
Subject: [Microscopy] viaWWW: The CAP checklist & ultramicrotomes

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Email: kenner.rita-at-marshfieldclinic.org
Name: Rita Kenner

Organization: Marshfield Clinic, Marshfield WI. 715-387-9159

Title-Subject: [Filtered] The CAP checklist & ultramicrotomes

Question: Greetings!
Does anyone out there have a Leica UC6 ultramicrotome? My lab recently bought one (I love it!), but now I have to rewrite my procedure in accordance with the CAP checklist question ANP.53000. My old ultramicrotome required a lengthy maintenance procedure of cleaning and lubricating parts etc etc. The UC6 is virtually maintenance-free (keep the dust and resin flakes wiped off, and clean the oculars & touch screen as needed).
How do you word this in your CAP procedure manual?
Thanks for enlightening me -
Rita Kenner
Electron Microscopist
Marshfield Clinic
Marshfield, WI 54449

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From: nizets2-at-yahoo.com
Date: Wed, 18 Jul 2007 03:14:58 -0500
Subject: [Microscopy] pipeting resin components

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html

Dear Paul,

Please forgive me for disturbing your feeling of
uniqueness ;-) but we also freeze large batch of resin
the same way or almost.
The only difference in that we aliquot the resin in
syringes, which are already calibrated. The syringes
are then closed with parafilm and frozen at -20°C.
Syringes are useful because you can use force and you
make not mistakes with the volumes (unlike pipetting)
and also there will be no remainings sticking to the
sides.
I must say that I don't even care about warming them
up in oven, I just take them out of the freezer and
the resin is fluid after 30 min R.T (just the time to
concentrate on your experiment while you prepare a cup
of coffee).
Hope it is useful.
Best regards,

Stephane


--- PWebster-at-hei.org wrote:

}
}
}
}
----------------------------------------------------------------------------
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} Microscopy Society of America
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}
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}
} Dear All,
}
} It seems that in many ways my lab may be unique in
} its way of handling
} resin. Many years ago I threw out the concept of
} solutions A and B , and the
} WPE values and went instead to a method first
} proposed to me by Sus Ito at
} Harvard.
}
} The method has worked consistently on three
} different continents and on a
} few EM course in different parts of the world. It
} consists of mixing three
} resin ingredients (but not the catalyst) in a large
} volume, aliquoting the
} mixture into small glass, screw-top vials and
} storing them, tightly closed,
} at -20 degrees for as long as it takes to finish the
} batch.
}
} As resin is needed, a vial containing from 4 to 6 ml
} of resin is removed
} from the freezer and warmed in the oven. The actual
} amount of resin in the
} vial can be determined by comparing the resin level
} with a calibrated (and
} marked) black vial). It is important to know the
} amount of resin in the
} vail because this will determine the amount of
} catalyst to use.
}
} Difficult to embed specimens can be soaked in the
} resin without catalyst for
} many days, or the resin can be mixed with catalyst
} and used immediately. The
} advantages are:
}
} 1. no messy resin preparation needed for each
} experiment.
} 2. no wasted resin components.
} 3. less handling of toxic components.
} 4. idiot-proof system ensuring reproducibility.
} 5. consistent results - the first vial is the same
} as the last vial.
} 6. no difficult measuring of catalyst (or any of the
} other components).
} 7. cleaner than other methods.
}
} Want to know the method? Here it is:
}
} Epon Resin (simplified recipe for making large
} batches of premixed resin)
}
} Mix the following in a large glass jar (see below):
}
} NMA 110 mL
} DDSA 130 mL
} Eponate 12 230 mL
}
} When the ingredients have been well mixed, the resin
} is poured into small
} tubes in aliquots of 4 to 6 mL and stored frozen.
} For use, warm a tube and,
} using a glass pasteur pipette, add four drops of
} BDMA per milliliter (or 2
} drops of DMP-30 if preferred). Mix well, use
} immediately and discard unused
} resin (polymerization is a good idea).
}
} Preparing and storing resin this way will ensure
} that resin ingredients are
} not wasted and all the tubes will have resin of the
} same consistency. The
} mixture can be tested before use by polymerizing one
} tube of resin. All
} subsequent tubes of resin will have the same
} polymerization and sectioning
} qualities as this first tube.
}
} Glass jar tip: A simple tip for measuring the resin
} components is to
} pre-calibrate a glass jar. In a clean jar, pour in
} 110mL of water, mark the
} meniscus. Add an extra 13mL of water and mark the
} new meniscus, and finally,
} add 230mL of water and mark the meniscus. (For those
} in the USA, Smuckers
} jars work well - UK, Robinson's orange marmalade -
} it just has to be clear
} glass!)
}
} Pour out all the water and dry the jar well. It is
} now ready to be used for
} measuring out the resin components and mixing them.
} Mix the three components
} well before pouring the resin into the small vials.
} As the volume is
} estimated just prior to the resin being used, there
} is no need to get into
} complicated pipetting methods, just pour carefully.
}
} As for glass pipettes (and I don't mean to be
} confrontational with the
} following comments), I have been using them for many
} years in my specimen
} preparation protocols. In addition to distributing
} catalyst, they are very
} useful for collecting scraped cells from culture
} dishes, and for
} transferring solutions from around very small
} specimens.
}
} I am aware that glass fragments are supposed to
} enter embedded specimen
} blocks and damage diamond knives, but most people I
} know also trim their
} specimen blocks using glass knives, which is a more
} reasonable source of
} glass contamination.
}
} In all the years I have been using glass pipettes I
} have never come across
} glass fragments in embedded tissues or cell pellets.
} I have seen hairs, dust
} and large chinks of keratin, but no glass. If glass
} were present in the
} blocks, I am sure it would be obvious in the initial
} trimming and facing
} off, which, incidentally we do with an old diamond
} knife.
}
} I send this method out for all to try. If anyone is
} brave enough to try it,
} let me know if it was useful - I promise that it
} will change your life,
} especially is you are part of a multi-user facility.
}
} Best regards,
}
} Paul Webster.
}
}
} Paul Webster, Ph.D
} House Ear Institute
} 2100 West Third Street
} Los Angeles, CA 90057
} (213) 273 8026
} pwebster-at-hei.org
}
}
}
}
}
}
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}
} I had to giggle at the many replies to my cry of
} frustration about
} measuring catalyst for resin! Like Tamara, over the
} years I have become
} adept at sequentially pouring the resin components
} into a tri-cornered
} beaker, stirring magnetically, then adding catalyst.
} Many years ago I used
} to measure out the components with syringes and then
} laboriously clean the
} syringes and keep them for the next use. Once I
} switched to weighing I
} kept using a tuberculin syringe to measure catalyst
} because my crappy
} balance couldn't weigh small amounts and did not do
} well in the fume
} hood; the draft shield was stolen years ago. Then I
} got mentally
} stuck. My syringes are desperately old, and the
} black rubber seal has
} been melting when in contact with the catalyst.
}
} Now when I train people they look at me like I'm
} crazy when I pour, so I
} let them transfer components to the beaker on the
} balance with disposable
}
=== message truncated ===




____________________________________________________________________________________
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From: edelmare-at-muohio.edu
Date: Wed, 18 Jul 2007 08:24:21 -0500
Subject: [Microscopy] Re: pipeting resin components

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html

Tina:

We weigh out all our resin components. And use a disposable pipet
to weigh out dropwise.

When working with a new resin mix for the first time if it does not
have weights (only volumes) we carefully weighout each
volumeterically measured component and right it down.

On 16 Jul 2007 at 19:43, tina-at-pbrc.hawaii.edu wrote:

}
}
}
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} ----------------------------------------------------------------------------
}
} Hi, all-
}
} Here's kind of a dumb question. How do you all measure out embedding resin
} catalyst? I've only been doing it for 31 years... I used to use tuberculin
} syringes - had to hurry because they sort of melt. The current crop of
} facility users seem dumbfounded that I would use anything but a
} pipetter. But I have now had another new pipetter get gummed up with some
} resin component, probably catalyst. Does anyone have a favorite method???
}
} Aloha with exasperation,
} Tina
}
} ****************************************************************************
} * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu *
} * Biological Electron Microscope Facility * (808) 956-6251 *
} * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf*
} ****************************************************************************
}
}
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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


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From: lcgould-at-med.cornell.edu
Date: Wed, 18 Jul 2007 09:29:58 -0500
Subject: [Microscopy] Re: pipeting resin components

Contents Retrieved from Microscopy Listserver Archives
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Dear Stephane,

You make no mention of when you add cataalyst to the resin. Do youincorporate ti into the final mixture that you freeze, or do you add it just before you use the resin?

There is a bg difference. If you add it to the resin before freezing, the resin will slowly harder, even at -20 degrees. This means that the properties of the resin are changing during storage. It also means the resin mixture can't be stored for long periods. We are able to store our uncatalysed resin for more than a year.

A possible complication of storing resin in plastic is that some plastics contain unincorporated plasticizer that could potentially inhibit, or otherwise alter the polymerization of the resin. Usually this is only a problem when setting up a new lab where the consumables are not exactly the same.

One usinque example of plastiziser interfering with polymerization is when polymerization of Lowicryl is attempted in new Eppendorf tubes. The new tubes have a volatile substace present that completely inhibits polymerization. The solution is to either autoclave the tubes, heat them in an oven overnight, or leave them in a cupboard for a few years.

My apologies for the errors in the last post, my finers refuse to learn how to spell.

Regards,

Paul.


House Ear Institute
2100 West Third Street
Los Angeles, CA 90057.

-----Original Message-----
X-from: Stephane Nizet [mailto:nizets2-at-yahoo.com]
Sent: Wed 7/18/2007 1:14 AM
To: Webster, Paul
Cc: microscopy-at-microscopy.com

In my earlier reply to Tina, I addressed only her immediate question:
how to measure out the catalyst. In response to Paul's posting, I do
a very similar thing, except that I use a method taught to me by my
first boss, the late Tom Robinson: I store the resin in the freezer,
in syringes. I cap the end (you can buy syringe caps from the
suppliers), and wrap the tips with parafilm. Works like a charm. If
you use Spurr's resin, you can even store it with the catalyst mixed
in for a few weeks at -20.
The syringes, obviously, make measuring the volume quite easy.
Lee
--
Leona Cohen-Gould, M.S.
Sr. Staff Associate
Director, Electron Microscopy & Histology Core Facility
Manager, Optical Microscopy Core Facility
Weill Cornell Medical College

voice (212)746-6146
fax (212)746-8175
http://www.cornellcelldevbiology.org
http://www.cornellbiochem.org

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From: rcommon-at-msu.edu
Date: Wed, 18 Jul 2007 09:42:28 -0500
Subject: [Microscopy] pipeting resin components

Contents Retrieved from Microscopy Listserver Archives
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My lab has been using a system similar to Dr. Webster's for 30 years. The
"Epon" resin components, minus the accelerator, are made up in batches of
200-400ml, thoroughly mixed and placed in glass vials in commonly used
aliquots (5,10,15,20,30,40ml). Some vials are left half filled to allow for
1:1 resin mixes. They are tightly sealed and stored at -20 degrees. Before
use the vials are allowed to come to room temperature and DMP is added. The
DMP is measured in disposable 1mm BD tuberculin syringes, without needles.
At about a nickel each, why wash glass syringes? As pointed out by others,
the resin can be stored much longer without the accelerator. And even
before resin stored with the accelerator added becomes unusable, it will be
partially polymerized and more viscous than it should be. We also use
disposable glass pipettes without problems.

Ralph Common
Michigan State University


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From: TindallR-at-missouri.edu
Date: Wed, 18 Jul 2007 09:55:59 -0500
Subject: [Microscopy] Pipetting resins: Question

Contents Retrieved from Microscopy Listserver Archives
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I am following this discussion with great interest, since I've heard
before about storing resins sans accelerator and adding this during the
final infiltration steps. We also store resins in the freezer, but we
use so much that it usually never has the chance to polymerize
significantly before it is used, so we store it with the accelerator
pre-mixed but there are times when it would be very convenient to make
huge resin batches or smaller batches of infrequently used resins, and
know that a month later they would still be predictably usable.

My questions are: regarding resins with and without the accelerator,
what are your infiltration schedules, and if the resin/accelerator mix
is added at the last step, do you always get consistent infiltration of
the accelerator into the tissue? In other words, isn't is possible to
get inconsistently hardened blocks if the the accelerator is added
during the last one or two changes of pure resin and doesn't get
completely and uniformly distributed in the tissue?

Thanks much,
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


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From: stefan.diller-at-t-online.de
Date: Wed, 18 Jul 2007 11:25:47 -0500
Subject: [Microscopy] Airfuge rotor

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html

Dear All,
I just bought a used Beckman Airfuge which came with this rotor:

www.elektronenmikroskopie.info/airfuge_rotor/

Can anybody tell me which rotor this is?
For what purposes it will normally be used?
What vials / inlays are needed?
The Airfuge spins up nicely with this rotor...

I would like to centrifuge virus particles on TEM grids. I understand, that
I may need another rotor. Which one would be best? Is a used one available
out there for "not so much" money?

Thanks,
Stefan


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From: Ian.Lamswood-at-leica-microsystems.com
Date: Wed, 18 Jul 2007 18:16:17 -0500
Subject: [Microscopy] viaWWW: Leica-Microsystems Job Opportunity

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Email: Ian.Lamswood-at-leica-microsystems.com
Name: Ian Lamswood

Organization: Leica Microsystems

Title-Subject: [Filtered] Job Opportunity

Question: Leica Microsystems, Vienna has an exciting opportunity in our Business Unit for a Product Manager.
Joining the team, you would be responsible for the complete life cycles of the cryo-instruments in our EM sample preparation product line. These include high pressure freezer and freeze substitution instruments.
The position is based in Vienna with international travel.

Experience
We are looking for someone with a biological background and commercial training, with a minimum of 3 years work experience in the areas of sales, marketing and/or product management. Knowledge of sample preparation equipment would be an advantage.
A readiness to travel is also required along with a good understanding of MS Office.
Fluent spoken and written English is essential.

If you are interested in this position please send your CV by e-mail to Otmar.Kases-at-leica-microsystems.com


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From: Elliott-at-arizona.edu
Date: Wed, 18 Jul 2007 20:35:06 -0500
Subject: [Microscopy] immuno-EM of Red Blood Cells

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html

Hi all
I am doing immuno-EM on human Red Blood Cells. In the past I have
used cryo-sections and probing on the grid. I would like to plastic
embed the cells and do antibody labeling on the grid. I used my
normal protocol and embedded in unicryl. Then my problem occurred.
The unicryl block polymerizes (UV for a few days at -10°C) just fine
except in the sample pellet. I assume the UV light is being absorbed
by the RBCs, and thus not penetrating the sample.

So, my question is, what should I try next?

Thank you
David




_____________________

David Elliott Ph.D.
Assistant Professor - Department of Cell Biology and Anatomy
Director, Research Microscopy Core Service
University of Arizona College of Medicine
PO Box 245004
Tucson, AZ 85724

Voice: 520-626-7870
Fax: 520-626-2097

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From: mmcheath-at-syr.edu
Date: Thu, 19 Jul 2007 08:54:04 -0500
Subject: [Microscopy] JEOL 733's and 8600's: position of vacuum gauge

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html

Hi,
Why is the vacuum gauge positioned at a right angle to the vertical tube
to the vacuum system on JEOL 733's and 8600's (and others in this line)?

Mike

********************************************************************
Michael M. Cheatham
312 Heroy Geology Laboratory Phone (315)-443-1261
Syracuse University Fax (315)-443-3363
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From: sougratr-at-mail.nih.gov
Date: Thu, 19 Jul 2007 09:05:20 -0500
Subject: [Microscopy] Re: immuno-EM of Red Blood Cells

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html

Hello David,

If the UV is the problem, you will solve it by using LR White resin with
the accelerator.
It will easily polymerizes at 4 degree using 1.5 microliter of
accelerator for 1 ml of resin.

Good luck.

Rachid

--
Rachid SOUGRAT
Cell Biology and Metabolism Branch
NICHD, NIH
Bldg. 18T, Rm. 101
18 Library Drive
Bethesda, MD 20892-5430
USA

Tel.: 301-594-3944
FAX 301-402-0078

http://rsougrat.googlepages.com/


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From: Elliott-at-arizona.edu
Date: Thu, 19 Jul 2007 11:37:49 -0500
Subject: [Microscopy] Re: immuno-EM of Red Blood Cells

Contents Retrieved from Microscopy Listserver Archives
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On Jul 19, 2007, at 5:25 AM, Philip Oshel wrote:

} David,
}
} First question: you don't mention osmium -- I suspect your samples
} are not osmicated, but just to be sure: are they?

They are not.

} Osmium interfers with polymerization of Unicryl and other acrylates.

It also interferes with antigenisity.

} If you're not, then I think you're right, the pellet is blocking
} the UV. I'd try dispersing the RBCs in agar, so the pellet isn't
} dense with cells.

The cells were dispersed in gelatin, but I could try making the cells
less concentrated.

} Otherwise: are you looking for a plasma membrane bound epitope? If
} yes, you should get good results with Au conjugated antibody
} labelling and FE-SEM, cryo or chemically fixed. Lot more to be seen
} this way, including patterns of distribution of the epitope on the
} membrane, something that's very difficult to find with sections.

No, I am looking at internal epitopes.

Thanx for the suggestions.

David
}
} Phil
}
} } ---------------------------------------------------------------------
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} } MicroscopyListserver
} } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} } ---------------------------------------------------------------------
} } -------
} }
} } Hi all
} } I am doing immuno-EM on human Red Blood Cells. In the past I have
} } used cryo-sections and probing on the grid. I would like to plastic
} } embed the cells and do antibody labeling on the grid. I used my
} } normal protocol and embedded in unicryl. Then my problem occurred.
} } The unicryl block polymerizes (UV for a few days at -10ƒC) just fine
} } except in the sample pellet. I assume the UV light is being absorbed
} } by the RBCs, and thus not penetrating the sample.
} }
} } So, my question is, what should I try next?
} }
} } Thank you
} } David
} }
} }
} }
} }
} } _____________________
} }
} } David Elliott Ph.D.
} } Assistant Professor - Department of Cell Biology and Anatomy
} } Director, Research Microscopy Core Service
} } University of Arizona College of Medicine
} } PO Box 245004
} } Tucson, AZ 85724
} }
} } Voice: 520-626-7870
} } Fax: 520-626-2097
} --
} Philip Oshel
} Microscopy Facility Supervisor
} Biology Department
} 024C Brooks Hall
} Central Michigan University
} Mt. Pleasant, MI 48859



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From: TindallR-at-missouri.edu
Date: Thu, 19 Jul 2007 12:03:19 -0500
Subject: [Microscopy] TEM: Bacteria in root nodules

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html

Dear Listers,

Holey Rhizobia, Batman!

We are running up against a deadline trying to image bacteria in root
nodules of a legume. Everything is working fine, except for the small
detail of myriads of holes in the resin wherever there are clusters of
bacteria.

First attempt was with Spurr's resin, using extended microwaving in the
processing and infiltration. By extended, I mean microwaving each
sample twice under vacuum at each infiltration step, followed by time on
a rocker (several hours to overnight) before going on to the next
infiltration step. This was a multi-day procedure. Holes, holes,
holes.

We next tried using LR White and an even more extended procedure (more
infiltration steps, more changes of pure resin) that has proven
successful at embedding whole retinas. Holes.......

The 100nm sections are on carbon-coated grids 200 mesh Cu grids.
Ultrastructure of the sample is fine and there are no holes anywhere
except where the bacteria are clustered, which, of course, is where we
need the pictures.

Any ideas? Anyone?

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


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From: connellyps-at-nhlbi.nih.gov
Date: Thu, 19 Jul 2007 13:09:17 -0500
Subject: [Microscopy] Re: TEM: Bacteria in root nodules

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html

Randy,

I had had problems with Listeria in monocytes back in the '80s. After much
grief of having holes inside the bacteria as well as peppering that formed
as the beam hit the bacterium (sorry to say published figures - the paper
was finished and went to press), I discovered that the cause was not only in
the infiltration, which I had already extended, but also in the dehydration.

The 'cure' was to increase time in all steps and add an additional exchange
of 100% EtOH (3 changes) for the bacteria were much more dense than the
surrounding tissue and tended to hold on to the moisture.

I was not using microwave so I do not know how much additional time would be
required for that technique.

I'm interested to see suggestions from others too.
Pat

Patricia Stranen Connelly
NHLBI Electron Microscopy Core
National Institutes of Health
14 Service Road South
Bldg. 14E ­ Rm. 111B MSC 5570
Bethesda, MD 20892-5570
Phone 301-496-3491
connellyps-at-mail.nih.gov


On 7/19/07 1:09 PM, "TindallR-at-missouri.edu" {TindallR-at-missouri.edu} wrote:

} Dear Listers,
}
} Holey Rhizobia, Batman!
}
} We are running up against a deadline trying to image bacteria in root
} nodules of a legume. Everything is working fine, except for the small
} detail of myriads of holes in the resin wherever there are clusters of
} bacteria.
}
} First attempt was with Spurr's resin, using extended microwaving in the
} processing and infiltration. By extended, I mean microwaving each
} sample twice under vacuum at each infiltration step, followed by time on
} a rocker (several hours to overnight) before going on to the next
} infiltration step. This was a multi-day procedure. Holes, holes,
} holes.
}
} We next tried using LR White and an even more extended procedure (more
} infiltration steps, more changes of pure resin) that has proven
} successful at embedding whole retinas. Holes.......
}
} The 100nm sections are on carbon-coated grids 200 mesh Cu grids.
} Ultrastructure of the sample is fine and there are no holes anywhere
} except where the bacteria are clustered, which, of course, is where we
} need the pictures.
}
} Any ideas? Anyone?
}
} 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
} ==============================End of - Headers==============================



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10, 24 -- Date: Thu, 19 Jul 2007 14:08:22 -0400
10, 24 -- Subject: Re: [Microscopy] TEM: Bacteria in root nodules
10, 24 -- From: Patricia Connelly {connellyps-at-nhlbi.nih.gov}
10, 24 -- To: Randy Tindall {TindallR-at-missouri.edu}
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From: PWebster-at-hei.org
Date: Thu, 19 Jul 2007 13:24:47 -0500
Subject: [Microscopy] immuno-EM of Red Blood Cells

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Dear David,

Fortunately you solution is easy - cut the gelatin embedded blocks of red blood cells into very thin slices and try your embedding again. The thinner slices will give more access tot he UV light and you will get the resin polymerized.

I have embedded gelatin-immobilized malaria-infected red blood cells in Lowicryl (polymerizing with UV light) on many occasions so I know the trick works.

Now, just don't ask me how to get rid of all the non-specific labeling over the haemoglobin!

Regards,

Paul.

House Ear Institute
2100 W 3rd St
Los Angeles
CA 90057


-----Original Message-----
X-from: Elliott-at-arizona.edu [mailto:Elliott-at-arizona.edu]
Sent: Wed 7/18/2007 6:39 PM
To: Webster, Paul

Hi all
I am doing immuno-EM on human Red Blood Cells. In the past I have
used cryo-sections and probing on the grid. I would like to plastic
embed the cells and do antibody labeling on the grid. I used my
normal protocol and embedded in unicryl. Then my problem occurred.
The unicryl block polymerizes (UV for a few days at -10°C) just fine
except in the sample pellet. I assume the UV light is being absorbed
by the RBCs, and thus not penetrating the sample.

So, my question is, what should I try next?

Thank you
David




_____________________

David Elliott Ph.D.
Assistant Professor - Department of Cell Biology and Anatomy
Director, Research Microscopy Core Service
University of Arizona College of Medicine
PO Box 245004
Tucson, AZ 85724

Voice: 520-626-7870
Fax: 520-626-2097

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From: brachfelds-at-mail.montclair.edu
Date: Thu, 19 Jul 2007 17:26:14 -0500
Subject: [Microscopy] AskAMicroscopist: Polarizing microscopes

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Email: brachfelds-at-mail.montclair.edu
Name: Stefanie Brachfeld

Organization: Montclair State University

Education: Graduate College

Location: City, State, Country

Title: Polarizing microscopes

Question: I am a faculty member purchasing a polarizing microscope for transmitted and reflected light analysis of geologic samples. I am new to light microscopy, and I am curious if anyone has experience and recommendations (performance, maintenance, frequency of repairs, quirks, etc.) on the different models of scopes that I am considering. They include the Leica DM2500, Zeiss Axioscope, Nikon LV100, and Olympus CX.
Thank you.

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From: kazmiruk-at-chtc.ru
Date: Thu, 19 Jul 2007 22:55:01 -0500
Subject: [Microscopy] confirmation

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Çäðàâñòâóéòå, Microscopy.



--
Ñ óâàæåíèåì,
Êàçüìèðóê mailto:kazmiruk-at-chtc.ru


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From: donc-at-asmicro.com
Date: Fri, 20 Jul 2007 01:04:30 -0500
Subject: [Microscopy] Re: [a] AskAMicroscopist: FEI Phenom Electron-light microscope -resolution

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Dear Foo Li,
You asked for confirmation of the stated 30 nm resolution of the FEI Phenom
table top SEM.

1) The general advice when buying any scientific instrument is that you
should confirm that it has the capabilities that you require for your work.
In this case, you should not rely on a nominal resolution figure that may
relate to materials very different from yours. So, I recommend you contact
the FEI Phenom group to arrange a demo on your materials.

2) I saw a demo of this unit at the Semicon West expo this week. The test
specimen is a 144-nm pitch, 2-dimensional grip of Aluminum bumps on Silicon.
The pattern was barely visible, with very poor contrast. In a good FE-SEM,
this pattern can easily be imaged. See
http://www.asmicro.com/Supplies/150-2D-Calibration-Specimen.htm and scroll
down to the SEM images.

3) Although the Phenom SEM could not make a useful image of the high
resolution pattern, I was impressed with the ease of use. They have
cleverly combined video OM imaging with a touch-screen display to provide
fingertip navigation on the specimen. Sample navigation is much more
natural here than in the Hitachi TM-1000, which made slightly better images
of the same pattern.

4) Disclaimer: I am not an expert on current SEM instrumentation. I'm an
AFM expert who uses SEM from time to time.You need to form your own opinion.
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: fli-at-estee.com
To: donc-at-asmicro.com
Sent: Tuesday, July 17, 2007 9:32 AM
Subject: [a] [Microscopy] AskAMicroscopist: FEI Phenom Electron-light
microscope does it





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Email: fli-at-estee.com
Name: Foo Li

Organization: Estee Lauder

Education: Graduate College

Location: Melville, NY 11747

Title: FEI Phenom Electron-light microscope

Question: Is the Phenom the real deal? 30 nm resolution is not all that
bad especially when dealing with emulsions. I need an opinion of this
product from actual Phenom users out there.

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From: dsherman-at-purdue.edu
Date: Fri, 20 Jul 2007 07:31:28 -0500
Subject: [Microscopy] Reference fo cryo-SEM

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Hi,

Does anyone have a general reference or review that is appropriate for
describing the technique of cryo-SEM? Here are lots of papers utilizing
cryo0SEM but I am looking for one that deals with introducing the technique
in addition to the resulting science.

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


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From: fab-at-tariffenet.it
Date: Fri, 20 Jul 2007 08:22:30 -0500
Subject: [Microscopy] rviaWWW: Light immunohistochemistry

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Email: fab-at-tariffenet.it
Name: Fabio

Organization: University of Catania

Title-Subject: [Filtered] Light immunohistochemistry

Question: Dear All,
I made an immunocytochemical reaction on paraffin section with HRP and DAB. Negative controls (just only without primary ab)are OK, but reaction slides show a diffuse and homoeneous cytoplasmic staining: What is the meaning of such results?

Thank You

fabio
UniCT

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From: phillipst-at-missouri.edu
Date: Fri, 20 Jul 2007 12:53:54 -0500
Subject: [Microscopy] QuickBoats

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I just saw something called "Quick Boats" on the EMS website. This is an
aluminum boat that clamps on to a glass knife with a set screw and silicon
rubber gasket. it is meant to be a reusable version of a Truf or metal tape
boat. Has anyone used one of these? Do they last with heavy use? Off-line
or Or-line responses welcomed. thanks, tom



Thomas E. Phillips, PhD
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400

573-882-4712 (office)
573-882-0123 (fax)
PhillipsT-at-missouri.edu
http://www.biology.missouri.edu/faculty/phillips.html
http://www.biotech.missouri.edu/mcc/


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From: nizets2-at-yahoo.com
Date: Fri, 20 Jul 2007 16:54:32 -0500
Subject: [Microscopy] Re: rviaWWW: Light immunohistochemistry

Contents Retrieved from Microscopy Listserver Archives
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Hi!

My first reaction would be to answer: your antigen has
a diffuse and homogeneous cytoplasmic pattern!

Now you must consider that HRP and DAB are enzymatic
reaction, which means that the product of the reaction
diffuses a bit. It is one of the major drawbacks of
this detection method. If you want precise
intracellular localisations, try immunofluorescence
and laser scanning confocal microscopy.

Best regards,

Stephane

--- fab-at-tariffenet.it wrote:

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}
} Email: fab-at-tariffenet.it
} Name: Fabio
}
} Organization: University of Catania
}
} Title-Subject: [Filtered] Light immunohistochemistry
}
} Question: Dear All,
} I made an immunocytochemical reaction on paraffin
} section with HRP and DAB. Negative controls (just
} only without primary ab)are OK, but reaction slides
} show a diffuse and homoeneous cytoplasmic staining:
} What is the meaning of such results?
}
} Thank You
}
} fabio
} UniCT
}
}
---------------------------------------------------------------------------
}
} ==============================Original
} Headers==============================
} 8, 11 -- From zaluzec-at-microscopy.com Fri Jul 20
} 08:22:29 2007
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} 8, 11 -- From: fab-at-tariffenet.it (by way of
} MicroscopyListserver)
} 8, 11 -- Subject: rviaWWW: Light
} immunohistochemistry
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}




____________________________________________________________________________________
Moody friends. Drama queens. Your life? Nope! - their life, your story. Play Sims Stories at Yahoo! Games.
http://sims.yahoo.com/

==============================Original Headers==============================
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10, 21 -- Subject: Re: [Microscopy] rviaWWW: Light immunohistochemistry
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From: dlowry-at-asu.edu
Date: Fri, 20 Jul 2007 17:36:25 -0500
Subject: [Microscopy] viaWWW: tannic acid

Contents Retrieved from Microscopy Listserver Archives
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Email: dlowry-at-asu.edu
Name: David Lowry

Organization: Arizona State University

Title-Subject: [Filtered] tannic acid

Question: Does tannic acid negatively affect antigenicity of epitopes for on-section immuno-labeling? I have used it previously during freeze-substitution for standard thin-section EM but am not aware of how it may affect labeling. If anyone may have a protocol or other advice they would be willing to share, it would be greatly appreciated.

---------------------------------------------------------------------------

==============================Original Headers==============================
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From: hainfeld-at-bnl.gov
Date: Fri, 20 Jul 2007 17:37:08 -0500
Subject: [Microscopy] viaWWW: gold labeling workshop

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Email: hainfeld-at-bnl.gov
Name: James F. Hainfeld

Organization: Brookhaven National Lab

Title-Subject: [Filtered] gold labeling workshop

Question: If you previously applied and have not been notified, please contact us again.

Gold Labeling Workshop
Sept. 11-13, 2007
Sponsored by the Brookhaven National Lab STEM (scanning transmission EM) Facility
A Hands-on Tutorial that will cover both theory and practice. Topics will include:
Gold nanoparticles: structure, synthesis, and properties
Molecular labeling strategies (to amines, thiols, genetic tags, active sites, complexes, nucleic acids, peptides)
Ni-NTA-gold
Sizes of gold from 0.8 nm to 30 nm
Isolation of labeled products (various chromatographies and other methods)
Unstained, negatively stained and Cryo samples
Comparison of STEM and TEM
Silver and gold enhancement
Image processing using gold labels
Problems and how to overcome them (e.g., labeling yield/occupancy, non-specific binding)
Other uses of gold nanoparticles (Western blots, light microscopy, medicine)
Laboratory participation to gold label a protein for EM
Whatís new

Limited Enrollment due to lab space: A course fee of $650 covers housing, meals (dorm, cafeteria), and materials.
Proposals to attend will be refereed: submit CV and letter describing you and your labís labeling interest.
Optionally describe a sample of yours that, if selected, would be labeled during the course.
Submit registration request to: Dr. James Hainfeld by email: hainfeld-at-bnl.gov
Brookhaven National Laboratory, Biology Dept., Bldg. 463, Upton, NY 11973, USA
Tel. 631-344-3372 * Fax

---------------------------------------------------------------------------


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From: ech-at-interchange.ubc.ca
Date: Sun, 22 Jul 2007 10:10:24 -0500
Subject: [Microscopy] viaWWW: M&M 2007 Family Affair

Contents Retrieved from Microscopy Listserver Archives
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Email: ech-at-interchange.ubc.ca
Name: Elaine Humphrey

Title-Subject: [Filtered] M&M 2007 Family Affair

Question: Dear all
Two years ago in Hawaii at the M&M meeting, Gracie Burke, then MSA President, initiated the "Family Affair" to make microscopy fun for the families who travelled with delegates of the conference and to give them an opportunity to visit the Exhibition floor. My talk was based around the theme of using microscopy to "Solve a Mystery." Since the next M&M is in Florida, it is expected that several families will be attending and we are extending the "Family Affair" to Monday and Tuesday afternoons.

The general theme of the talks will again be "Solve the Mystery" (different mysteries from Hawaii) using microscopy with a biological emphasis on Monday and a materials emphasis on Tuesday.

Since the convention center is located in the Port of Ft. Lauderdale, an ID must be shown for access to the port. Anyone can enter the port, then the convention center, after showing an ID. This is a reminder to make sure that when you bring the family to the convention centre they should all have IDs with them. This is not a problem for those families arriving by air, but families driving to Ft. Lauderdale may not think to pack IDs for the kids.

best wishes
Elaine



---------------------------------------------------------------------------

==============================Original Headers==============================
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10, 11 -- To: microscopy-at-microscopy.com
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10, 11 -- Subject: viaWWW: M&M 2007 Family Affair
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From: Gerd.Leitinger-at-meduni-graz.at
Date: Mon, 23 Jul 2007 03:51:58 -0500
Subject: [Microscopy] TEM: lateral resolution immunogold

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html

Dear list

I am wondering whether anybody knows a /citation /for the lateral
resolution of immunogold stains.
I have ± 20nm in my head (this should correspond to the lenth of two
antibody molecules if there is one primary and one secondary antibody
between the gold granules and the epitope) and don't know whether I read
this from a posting a while ago or in an article.

thank you - as always

Gerd


Dr Gerd Leitinger
Institute of Cell Biology, JHistology and Embryology
Medical University of Graz, Austria


==============================Original Headers==============================
7, 18 -- From Gerd.Leitinger-at-meduni-graz.at Mon Jul 23 03:51:58 2007
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From: stmccart-at-vt.edu
Date: Mon, 23 Jul 2007 08:47:12 -0500
Subject: [Microscopy] web based instrument sign up

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html

Hello: We are going to a web based sign up for a new characterization
facility that will contain about 10 instruments. We have done a search of
the archives and found 3 systems, FACES and one from Northwestern and U. of
Arizona. Are there others that people are using we should
consider. Thanks. Steve

Stephen McCartney
Senior Research Associate
Nanoscale Characterization and Fabrication Laboratory (NCFL)
Institute for Critical Technology and Applied Science (ICTAS)
1991 Kraft Drive
Va Tech
Blacksburg, VA 24061
540-231-9765


==============================Original Headers==============================
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3, 20 -- From: Stephen McCartney {stmccart-at-vt.edu}
3, 20 -- Subject: web based instrument sign up
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==============================End of - Headers==============================




From: zaluzec-at-aaem.amc.anl.gov
Date: Mon, 23 Jul 2007 09:13:17 -0500
Subject: [Microscopy] Re: web based instrument sign up

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


Stephen

At the ANL EM Center we use a commerical web based calendar system
for all our instruments, called Calcium. http://www.brownbearsw.com/

It is not designed for microscopy, but it works quite well for our needs.

You can see the on-line version of how we use this at the following URL

http://emc.msd.anl.gov/cgi-bin/Calcium39.pl

Since you do not have an account all you can do is view the calendar.

Neither I nor ANL have any financial interests in this software, other
than being users.

Nestor
Your Friendly Neighborhood SysOp


} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America


--
===========================================
Dr. Nestor J. Zaluzec
Argonne National Lab
Electron Microscopy Center
Materials Science Division/Bldg 212
9700 S. Cass Ave
Argonne, Illinois 60439 USA
Tel: 630-252-7901, Fax: 630-252-4798
Email: Zaluzec-at-aaem.amc.anl.gov
===========================================
TPMLab: http://tpm.amc.anl.gov
MMSite: http://www.amc.anl.gov
===========================================

The box said ...
"This program requires Win 95/98/NT or better..."
So I bought a Mac !

===========================================

==============================Original Headers==============================
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15, 16 -- Date: Mon, 23 Jul 2007 09:13:11 -0500
15, 16 -- To: microscopy-at-microscopy.com
15, 16 -- From: "Nestor J. Zaluzec" {zaluzec-at-aaem.amc.anl.gov}
15, 16 -- Subject: Re: [Microscopy] web based instrument sign up
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==============================End of - Headers==============================




From: dsherman-at-purdue.edu
Date: Mon, 23 Jul 2007 09:56:43 -0500
Subject: [Microscopy] web based instrument sign up

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html

Does anyone know of a calendar program appropriate for a microscopy lab that
will run on the MAC?

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


} From: {zaluzec-at-aaem.amc.anl.gov}
} Reply-To: {zaluzec-at-aaem.amc.anl.gov}
} Date: Mon, 23 Jul 2007 09:17:00 -0500
} To: {dsherman-at-purdue.edu}
} Subject: [Microscopy] Re: web based instrument sign up
}
}
}
}
} ----------------------------------------------------------------------------
} 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
} ----------------------------------------------------------------------------
}
}
} Stephen
}
} At the ANL EM Center we use a commerical web based calendar system
} for all our instruments, called Calcium. http://www.brownbearsw.com/
}
} It is not designed for microscopy, but it works quite well for our needs.
}
} You can see the on-line version of how we use this at the following URL
}
} http://emc.msd.anl.gov/cgi-bin/Calcium39.pl
}
} Since you do not have an account all you can do is view the calendar.
}
} Neither I nor ANL have any financial interests in this software, other
} than being users.
}
} Nestor
} Your Friendly Neighborhood SysOp
}
}
} } ----------------------------------------------------------------------------
} } 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: We are going to a web based sign up for a new characterization
} } facility that will contain about 10 instruments. We have done a search of
} } the archives and found 3 systems, FACES and one from Northwestern and U. of
} } Arizona. Are there others that people are using we should
} } consider. Thanks. Steve
} }
} } Stephen McCartney
} } Senior Research Associate
} } Nanoscale Characterization and Fabrication Laboratory (NCFL)
} } Institute for Critical Technology and Applied Science (ICTAS)
} } 1991 Kraft Drive
} } Va Tech
} } Blacksburg, VA 24061
} } 540-231-9765
} }
} }
} } ==============================Original Headers==============================
} } 3, 20 -- From stmccart-at-vt.edu Mon Jul 23 08:47:11 2007
} } 3, 20 -- Received: from lennier.cc.vt.edu (lennier.cc.vt.edu
} } [198.82.162.213])
} } 3, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with
} } ESMTP id l6NDlBen001511
} } 3, 20 -- for {microscopy-at-microscopy.com} ; Mon, 23 Jul 2007
} } 08:47:11 -0500
} } 3, 20 -- Received: from vivi.cc.vt.edu
} } (IDENT:mirapoint-at-evil-vivi.cc.vt.edu [10.1.1.12])
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} } ESMTP id l6NDlAZL019144
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} } 09:47:10 -0400
} } 3, 20 -- Received: from mri-system.vt.edu ([198.82.148.26])
} } 3, 20 -- by vivi.cc.vt.edu (MOS 3.8.2-GA)
} } 3, 20 -- with ESMTP id HTD69749;
} } 3, 20 -- Mon, 23 Jul 2007 09:47:10 -0400 (EDT)
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} } 3, 20 -- X-Sender: stmccart-at-pop.vt.edu
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} } 3, 20 -- Date: Mon, 23 Jul 2007 09:46:58 -0400
} } 3, 20 -- To: Microscopy Listserver {microscopy-at-microscopy.com}
} } 3, 20 -- From: Stephen McCartney {stmccart-at-vt.edu}
} } 3, 20 -- Subject: web based instrument sign up
} } 3, 20 -- Mime-Version: 1.0
} } 3, 20 -- Content-Type: text/plain; charset="us-ascii"; format=flowed
} } ==============================End of - Headers==============================
}
}
} --
} ===========================================
} Dr. Nestor J. Zaluzec
} Argonne National Lab
} Electron Microscopy Center
} Materials Science Division/Bldg 212
} 9700 S. Cass Ave
} Argonne, Illinois 60439 USA
} Tel: 630-252-7901, Fax: 630-252-4798
} Email: Zaluzec-at-aaem.amc.anl.gov
} ===========================================
} TPMLab: http://tpm.amc.anl.gov
} MMSite: http://www.amc.anl.gov
} ===========================================
}
} The box said ...
} "This program requires Win 95/98/NT or better..."
} So I bought a Mac !
}
} ===========================================
}
} ==============================Original Headers==============================
} 15, 16 -- From zaluzec-at-aaem.amc.anl.gov Mon Jul 23 09:13:16 2007
} 15, 16 -- Received: from aaem.amc.anl.gov (aaem.amc.anl.gov [146.139.72.3])
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} 15, 16 -- From: "Nestor J. Zaluzec" {zaluzec-at-aaem.amc.anl.gov}
} 15, 16 -- Subject: Re: [Microscopy] web based instrument sign up
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==============================Original Headers==============================
6, 23 -- From dsherman-at-purdue.edu Mon Jul 23 09:56:43 2007
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6, 23 -- Date: Mon, 23 Jul 2007 10:56:41 -0400
6, 23 -- Subject: Re: [Microscopy] Re: web based instrument sign up
6, 23 -- From: Debby Sherman {dsherman-at-purdue.edu}
6, 23 -- To: {zaluzec-at-aaem.amc.anl.gov} ,
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From: gmartens-at-interchange.ubc.ca
Date: Mon, 23 Jul 2007 10:07:00 -0500
Subject: [Microscopy] Re: web based instrument sign up

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html

Hi Debby and Steve,

Calcium will also run on a Mac based platform.

We have been running Calcium for several years now and have been very
happy with the software.

--
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==============================
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6, 29 -- From: Garnet Martens {gmartens-at-interchange.ubc.ca}
6, 29 -- Subject: Re: [Microscopy] web based instrument sign up
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From: jpchandl-at-mines.edu
Date: Mon, 23 Jul 2007 10:32:50 -0500
Subject: [Microscopy] Re: web based instrument sign up

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html

We also wanted to move from a paper based reservation system to an online
one. The calcium program that Nestor and others have mentioned is used by a
lot of satisfied customers, and we almost became one of them. It was at the
top of our list for purchase.

When we discussed the issues surrounding installation and maintenance of
such a program with our network services staff, we found that we already had
an online calendar on campus that might do what we wanted. It is part of
the Banner suite of software packages from SCT, and includes the campus-wide
finance, student records and campus portal system. The portal has a
calendar function that we could use.

Calendars for each instrument can be created by the portal administrator,
and then maintenance is up to people in the lab. Maintenance is simple and
straight forward and is web based. Signup is also web based, so checking
instrument availability and signup can be done from most anywhere. One of
the vendor's intended uses of this calendar is to set up meetings, so signup
consists of what amounts to setting up a meeting between yourself and an
instrument. Also, signup requires login privileges to the campus portal
system.

This is a simple system and does not have some features that might be
important to some labs, like logging of changes, restrictions to specific
times, maximum signup time allowed, etc. It simply replaced our paper
calendars, which were pretty low tech. It was also already available, and,
therefore, free.

--John
John Chandler, Ph.D.
Manager, Electron Microscopy Lab
Colorado School of Mines
1500 Illinois Street
Golden, CO 80401-1887
jpchandl-at-mines.edu
303.384.2203

-----Original Message-----
X-from: zaluzec-at-aaem.amc.anl.gov [mailto:zaluzec-at-aaem.amc.anl.gov]
Sent: Monday, July 23, 2007 8:20 AM
To: jpchandl-at-mines.edu


Stephen

At the ANL EM Center we use a commerical web based calendar system
for all our instruments, called Calcium. http://www.brownbearsw.com/

It is not designed for microscopy, but it works quite well for our needs.

You can see the on-line version of how we use this at the following URL

http://emc.msd.anl.gov/cgi-bin/Calcium39.pl

Since you do not have an account all you can do is view the calendar.

Neither I nor ANL have any financial interests in this software, other
than being users.

Nestor
Your Friendly Neighborhood SysOp


} ---------------------------------------------------------------------------
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} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America



==============================Original Headers==============================
22, 21 -- From jpchandl-at-mines.edu Mon Jul 23 10:32:50 2007
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22, 21 -- References: {200707231420.l6NEKPZc023433-at-ns.microscopy.com}
22, 21 -- Subject: RE: [Microscopy] Re: web based instrument sign up
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==============================End of - Headers==============================




From: thoward-at-unm.edu
Date: Mon, 23 Jul 2007 11:36:46 -0500
Subject: [Microscopy] Re: web based instrument sign up

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html

The calendar function in Groupwise can also be used to
reserve equipment. We don't use it for the 'scopes, but
the shared real-time PCR machines are scheduled this way.
If it is already in place at your institution, you'd be
ready to roll.

Tamara

On Mon, 23 Jul 2007 08:50:01 -0500
stmccart-at-vt.edu wrote:
}
}
}
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} Hello: We are going to a web based sign up for a new
} characterization
} facility that will contain about 10 instruments. We
} have done a search of
} the archives and found 3 systems, FACES and one from
} Northwestern and U. of
} Arizona. Are there others that people are using we
} should
} consider. Thanks. Steve
}
} Stephen McCartney
} Senior Research Associate
} Nanoscale Characterization and Fabrication Laboratory
} (NCFL)
} Institute for Critical Technology and Applied Science
} (ICTAS)
} 1991 Kraft Drive
} Va Tech
} Blacksburg, VA 24061
} 540-231-9765
}
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} ==============================Original
} Headers==============================
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***************************
Tamara Howard
Cell Biology & Physiology
UNM-HSC
Albuquerque, NM
***************************

==============================Original Headers==============================
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From: toli-at-hillweb.com
Date: Mon, 23 Jul 2007 12:23:50 -0500
Subject: [Microscopy] ||| Microscopes |||

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html

We are in the process of upgrading our microscopes and are out of space.
The scopes listed bellow are available for sale.

*-------------------------------------------------------------------------------------------------*
Images of the scopes can be downloaded (PDF) at:

http://www.3dfs.com/scopes_BioWarn.pdf

-- If you are having a hard time viewing the pdf, please email me:
toli-at-hillweb.com*
-------------------------------------------------------------------------------------------------*
*
*
*Jeol JSM-35C*
Working SEM, has been used as digital scope with A/D card (not included).
Includes Link Analytical backscattering detector.
12k OBO

-------------------------------------------------------------------------------------------------

*Zeiss EM-10*
Working TEM, fantastic resolution, includes Scion digital camera
Included is a spare microscope for parts
17k OBO

-------------------------------------------------------------------------------------------------

*ISI-40 SEM*
Working condition
9k OBO
-------------------------------------------------------------------------------------------------
*
**Jeol 100-S TEM*
Lots of work done to restore machine, but has not been turned
on recently
5k OBO

-------------------------------------------------------------------------------------------------

- Microscopes professionally maintained.
- Microscopes located in Pittsboro North Carolina, USA
- We have means for professionally shutting down/ packaging and shipping
the scopes.

-------------------------------------------------------------------------------------------------
*For technical questions please contact:*
Dr. Wolfgang Wagner, Director of microscopy
919-542-7410
wagner-at-hillweb.com*
*
*For sales please contact:*
¦ Anatoli Oleynik, Research Associate - BioWarn, LLC.
¦ 964 East St. Suite 106a - Pittsboro, NC 27312
¦ O: 919.542.7410 | M: 919.260.1911 | F: 919.542.0268
¦ mobile email/MSN-M: cell.toli-at-hotmail.com
¦ www.biowarnllc.com
*
*

==============================Original Headers==============================
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From: sbarlow-at-sunstroke.sdsu.edu
Date: Mon, 23 Jul 2007 12:26:00 -0500
Subject: [Microscopy] Re: web based instrument sign up

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html

Our University maintains a web-based group meeting calender,
MeetingMaker, which we have used for about 3 years without a problem.
(I don't have a financial interest in this software or company)

The advantages of this setup are it is very easy to use, and the
University does the housekeeping chores vis-a-vis security, hackers,
updates, etc.

The disadvantage is the system is only useful for reserving time on
equipment. It isn't useful for linking to the equipment and
automatically recording actual user time on equipment, assimilating
the information on some sort of weekly or monthly schedule to a
common location, making and posting billing, then collecting the
money.

I know a number of us would like a system that does more than just
reserve time on a calender.

Steve

} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America

--
Dr. Steven Barlow
EM Facility/Biology Dept.
San Diego State University
5500 Campanile Drive
San Diego CA 92182-4614
phone: (619) 594-4523
fax: (619) 594-5676

email: sbarlow-at-sunstroke.sdsu.edu
http://www.sci.sdsu.edu/emfacility

==============================Original Headers==============================
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From: jmkrupp-at-ucsc.edu
Date: Mon, 23 Jul 2007 12:55:37 -0500
Subject: [Microscopy] ISI WB-6 repair?

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html

Hi:

Our vintage ISI WB-6 has suffered what appears to be a catastrophic failure.

It can't seem to focus in the high mag range. Everything acts OK, but
the image will not focus, either autofocus or manual.

Our indy service guy has taken a look and thinks it might be a short
in the obj lens, requiring a major dismantle to get in there to check
it, and if it is, then what?. With no access to anything like factory
service, does anyone have any ideas?

My dilemma is that I have a Hitachi S-2700 that I picked up for free
waiting to be installed. Not sure if the Hitachi will work, was not
operational when I picked it up, but it looks almost brand new. Price
for the repair or the new install will be about the same. Which of
course, is money that I don't have right now, but I am trying to plan
ahead.

Before declaring the ISI a total loss and rolling it into the
dumpster, anyone have a suggestion about a fix?

Thanks

Jon
--

Jonathan Krupp
Microscopy & Imaging Lab
C230 Earth & Marine Science
University of California
Santa Cruz, CA 95064
(831) 459-2477
jmkrupp-at-ucsc.edu

I rode in AIDS/Lifecycle 6 SF to LA June 3 - 9, 2007 to raise money
for the San Francisco AIDS Foundation. Visit
http://www.aidslifecycle.org for more information about the ride or
http://www.aidslifecycle.org/5482 to make a donation.

==============================Original Headers==============================
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10, 17 -- Subject: ISI WB-6 repair?
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==============================End of - Headers==============================




From: bbandli-at-mvainc.com
Date: Mon, 23 Jul 2007 13:46:15 -0500
Subject: [Microscopy] web based instrument sign up

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html

Has anyone played around with using Google Calendar for scheduling
instrument time? I use it to manage my daily agenda, but don't have a
need (yet) to schedule instrument time, but it looks like a possible
solution. I am not at all familiar with any other scheduling programs
so Google Calendar may not be advanced enough, but since it's free and
works with pretty much any browser it might be a solution.

--
-----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------
Bryan Bandli
Research Microscopist
MVA Scientific Consultants
3300 Breckinridge Blvd., Suite 400
(770) 662-8509
bbandli-at-mvainc.com
www.mvainc.com
-----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------
The information in this email is confidential and may be legally
privileged.
It is intended solely for the addressee. If you are not the intended
recipient, please delete the email and notify MVA Scientific Consultants
of the transmission error.
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==============================Original Headers==============================
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From: Elliott-at-arizona.edu
Date: Mon, 23 Jul 2007 13:53:26 -0500
Subject: [Microscopy] Re: web based instrument sign up

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html

OneCore (http://demo.arl.arizona.edu/) will run on a Mac.
We use the code. Does fine.
David


On Jul 23, 2007, at 8:01 AM, dsherman-at-purdue.edu wrote:

}
}
}
} ----------------------------------------------------------------------
} ------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of
} America
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} MicroscopyListserver
} On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} ----------------------------------------------------------------------
} ------
}
} Does anyone know of a calendar program appropriate for a microscopy
} lab that
} will run on the MAC?
}
} 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
}
}
} } From: {zaluzec-at-aaem.amc.anl.gov}
} } Reply-To: {zaluzec-at-aaem.amc.anl.gov}
} } Date: Mon, 23 Jul 2007 09:17:00 -0500
} } To: {dsherman-at-purdue.edu}
} } Subject: [Microscopy] Re: web based instrument sign up
} }
} }
} }
} }
} } ---------------------------------------------------------------------
} } -------
} } 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
} } ---------------------------------------------------------------------
} } -------
} }
} }
} } Stephen
} }
} } At the ANL EM Center we use a commerical web based calendar system
} } for all our instruments, called Calcium. http://www.brownbearsw.com/
} }
} } It is not designed for microscopy, but it works quite well for our
} } needs.
} }
} } You can see the on-line version of how we use this at the
} } following URL
} }
} } http://emc.msd.anl.gov/cgi-bin/Calcium39.pl
} }
} } Since you do not have an account all you can do is view the calendar.
} }
} } Neither I nor ANL have any financial interests in this software,
} } other
} } than being users.
} }
} } Nestor
} } Your Friendly Neighborhood SysOp
} }
} }
} } } --------------------------------------------------------------------
} } } --------
} } } 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: We are going to a web based sign up for a new
} } } characterization
} } } facility that will contain about 10 instruments. We have done a
} } } search of
} } } the archives and found 3 systems, FACES and one from Northwestern
} } } and U. of
} } } Arizona. Are there others that people are using we should
} } } consider. Thanks. Steve
} } }
} } } Stephen McCartney
} } } Senior Research Associate
} } } Nanoscale Characterization and Fabrication Laboratory (NCFL)
} } } Institute for Critical Technology and Applied Science (ICTAS)
} } } 1991 Kraft Drive
} } } Va Tech
} } } Blacksburg, VA 24061
} } } 540-231-9765
} } }
} } }
} } } ==============================Original
} } } Headers==============================
} } } 3, 20 -- From stmccart-at-vt.edu Mon Jul 23 08:47:11 2007
} } } 3, 20 -- Received: from lennier.cc.vt.edu (lennier.cc.vt.edu
} } } [198.82.162.213])
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} } } 3, 20 -- Date: Mon, 23 Jul 2007 09:46:58 -0400
} } } 3, 20 -- To: Microscopy Listserver {microscopy-at-microscopy.com}
} } } 3, 20 -- From: Stephen McCartney {stmccart-at-vt.edu}
} } } 3, 20 -- Subject: web based instrument sign up
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} } } 3, 20 -- Content-Type: text/plain; charset="us-ascii"; format=flowed
} } } ==============================End of -
} } } Headers==============================
} }
} }
} } --
} } ===========================================
} } Dr. Nestor J. Zaluzec
} } Argonne National Lab
} } Electron Microscopy Center
} } Materials Science Division/Bldg 212
} } 9700 S. Cass Ave
} } Argonne, Illinois 60439 USA
} } Tel: 630-252-7901, Fax: 630-252-4798
} } Email: Zaluzec-at-aaem.amc.anl.gov
} } ===========================================
} } TPMLab: http://tpm.amc.anl.gov
} } MMSite: http://www.amc.anl.gov
} } ===========================================
} }
} } The box said ...
} } "This program requires Win 95/98/NT or better..."
} } So I bought a Mac !
} }
} } ===========================================
} }
} } ==============================Original
} } Headers==============================
} } 15, 16 -- From zaluzec-at-aaem.amc.anl.gov Mon Jul 23 09:13:16 2007
} } 15, 16 -- Received: from aaem.amc.anl.gov (aaem.amc.anl.gov
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} } 15, 16 -- Date: Mon, 23 Jul 2007 09:13:11 -0500
} } 15, 16 -- To: microscopy-at-microscopy.com
} } 15, 16 -- From: "Nestor J. Zaluzec" {zaluzec-at-aaem.amc.anl.gov}
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}
} ==============================Original
} Headers==============================
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} 6, 23 -- User-Agent: Microsoft-Entourage/11.3.6.070618
} 6, 23 -- Date: Mon, 23 Jul 2007 10:56:41 -0400
} 6, 23 -- Subject: Re: [Microscopy] Re: web based instrument sign up
} 6, 23 -- From: Debby Sherman {dsherman-at-purdue.edu}
} 6, 23 -- To: {zaluzec-at-aaem.amc.anl.gov} ,
} 6, 23 -- "message to: MSA list" {microscopy-at-microscopy.com}
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==============================Original Headers==============================
5, 22 -- From Elliott-at-arizona.edu Mon Jul 23 13:53:25 2007
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5, 22 -- From: David Elliott {Elliott-at-arizona.edu}
5, 22 -- Subject: Re: [Microscopy] web based instrument sign up
5, 22 -- Date: Mon, 23 Jul 2007 11:51:25 -0700
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From: Sally.Stowe-at-anu.edu.au
Date: Mon, 23 Jul 2007 17:35:31 -0500
Subject: [Microscopy] Re: web based instrument sign up, calendar,

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html

Hi,
we also use the google calendar for keeping track of what is going on
within the lab - courses, staff movements, seminars etc, but for
instrument booking and simple usage records we use a home-grown SQL-based
system - it was made several years ago by Nathan Cattle, an engineering
student now in IT support, which is handy...we can get upgrades and
improvements from time to time. We have users outside our institution, so
can't use an institution-based system.

You can see a users-eye view of some equipment (SEMs) by going to
http://www.anu.edu.au/EMU/booking/ and using DisplayS as both login and
password. DisplayL , DisplayT..you get the picture. These are dummy users
set up for convenience..but please dont actually make a booking!

Administrators (anyone can be set up temporarily as an administrator)can
quickly set up new users with varying levels of access, new departments,
and new instruments. Instruments have a range of usage patterns, and
maximum bookable times. Major accessories or configuration options can be
flagged. Staff concerned with an instrument optionally receive emails when
a booking is made, with any comments entered by the user. Users tagged as
"needing assistance" can easily be contacted to confirm or refuse
bookings. Staff have a list of bookings for which they are responsible on
their home page.

Stats of individual bookings (not filament time) are stored and can be
accessed by instrument and month..this might be extended later. Users only
see items they can access.


It works well for us, for about 25 instruments, and other departments have
piggy-backed on the system for another 15 or so. This works without
cluttering the screens because most users and administrators are not shown
anything they dont need. So a molecular biology lab down the hall or in
another building can operate effectively as an independent system.

Contact Nathan if you want to know more.

On a related note, we have just launched DOSSER
http://dosser.anu.edu.au/index.php, a directory for "accessible"
equipment. You dont need to sign up to browse or search it, only to add
new equipment. Comments welcome!

cheers
Sally


Dr SJ Stowe
Facility Coordinator
ANU Electron Microscopy Unit
www.anu.edu.au/EMU/

On Mon, July 23, 2007 11:47 pm, stmccart-at-vt.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
} --------------------------------------------------------------------------
} --
}
}
} Hello: We are going to a web based sign up for a new characterization
} facility that will contain about 10 instruments. We have done a search of
} the archives and found 3 systems, FACES and one from Northwestern and U.
} of Arizona. Are there others that people are using we should
} consider. Thanks. Steve
}
} Stephen McCartney
} Senior Research Associate
} Nanoscale Characterization and Fabrication Laboratory (NCFL)
} Institute for Critical Technology and Applied Science (ICTAS)
} 1991 Kraft Drive
} Va Tech
} Blacksburg, VA 24061
} 540-231-9765
}






==============================Original Headers==============================
18, 27 -- From sally.stowe-at-anu.edu.au Mon Jul 23 17:35:31 2007
18, 27 -- Received: from mail.rsbs.anu.edu.au (mail.rsbs.anu.edu.au [150.203.72.70])
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18, 27 -- Date: Tue, 24 Jul 2007 08:35:23 +1000 (EST)
18, 27 -- Subject: Re: [Microscopy] web based instrument sign up, calendar,
18, 27 -- DOSSER for shared equipment
18, 27 -- From: Sally.Stowe-at-anu.edu.au
18, 27 -- To: stmccart-at-vt.edu, microscopy-at-microscopy.com
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From: dang-at-oakland.edu
Date: Mon, 23 Jul 2007 18:36:37 -0500
Subject: [Microscopy] AskAMicroscopist: METHOD TO PROCESS THE LEAF FOR TEM

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html

This Question was submitted to Ask-A-Microscopist by (dang-at-oakland.edu)
from http://www.microscopy.org/Ask-A-Microscopist/Ask-A-Microscopist.html on Monday, July 23, 2007 at 13:42:20
Remember to consider the Grade/Age of the student when considering the Question
---------------------------------------------------------------------------
Please reply to both dang-at-oakland.edu as well as to the Microscopy Listserver
---------------------------------------------------------------------------

Email: dang-at-oakland.edu
Name: LOAN DANG

Organization: Oakland University

Education: Graduate College

Location: Rochester Hills michigan 48309

Title: METHOD TO PROCESS THE LEAF FOR TEM

Question: I would like to have a protocol to fix the leaf
and process it for the TEM
Thank you
LOAN DANG

---------------------------------------------------------------------------

==============================Original Headers==============================
8, 11 -- From zaluzec-at-ultra5.microscopy.com Mon Jul 23 18:36:37 2007
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8, 11 -- From: dang-at-oakland.edu (by way of Ask-A-Microscopist)
8, 11 -- Subject: AskAMicroscopist: METHOD TO PROCESS THE LEAF FOR TEM
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From: syli-at-northwestern.edu
Date: Mon, 23 Jul 2007 18:46:15 -0500
Subject: [Microscopy] Re: web based instrument sign up, calendar,

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html


Hi,

The advantages of a dedicated software over general web calendar systems
are:

1. It allows convenient tracking of instrument usage/repair history.
2. It gives statistical reports by departments, by PIs, or by instruments,
which are important for funding applications and beyond.
3. It generates billing spreadsheet with several mouse clicks only.
4. It usually integrates solid access controls (remote relays) to the
instruments, so users cannot use unauthorized instruments without logging
in.

All these functions are critical for a recharge facility. However for simple
share of instruments without accounting needs, web calendar system should be
sufficient.

Shuyou
_____________________________
Shuyou Li, Ph.D.
NUANCE Center
Northwestern University
2220 Campus Drive, 2036 Cook Hall
Evanston, IL 60208-3108, USA
Ph: (847) 491-6723
Fax: (847) 467-6573
Email: syli-at-northwestern.edu
http://www.nuance.northwestern.edu/


-----Original Message-----
X-from: Sally.Stowe-at-anu.edu.au [mailto:Sally.Stowe-at-anu.edu.au]
Sent: Monday, July 23, 2007 5:42 PM
To: syli-at-northwestern.edu

Hi,
we also use the google calendar for keeping track of what is going on within
the lab - courses, staff movements, seminars etc, but for instrument booking
and simple usage records we use a home-grown SQL-based system - it was made
several years ago by Nathan Cattle, an engineering student now in IT
support, which is handy...we can get upgrades and improvements from time to
time. We have users outside our institution, so can't use an
institution-based system.

You can see a users-eye view of some equipment (SEMs) by going to
http://www.anu.edu.au/EMU/booking/ and using DisplayS as both login and
password. DisplayL , DisplayT..you get the picture. These are dummy users
set up for convenience..but please dont actually make a booking!

Administrators (anyone can be set up temporarily as an administrator)can
quickly set up new users with varying levels of access, new departments, and
new instruments. Instruments have a range of usage patterns, and maximum
bookable times. Major accessories or configuration options can be flagged.
Staff concerned with an instrument optionally receive emails when a booking
is made, with any comments entered by the user. Users tagged as "needing
assistance" can easily be contacted to confirm or refuse bookings. Staff
have a list of bookings for which they are responsible on their home page.

Stats of individual bookings (not filament time) are stored and can be
accessed by instrument and month..this might be extended later. Users only
see items they can access.


It works well for us, for about 25 instruments, and other departments have
piggy-backed on the system for another 15 or so. This works without
cluttering the screens because most users and administrators are not shown
anything they dont need. So a molecular biology lab down the hall or in
another building can operate effectively as an independent system.

Contact Nathan if you want to know more.

On a related note, we have just launched DOSSER
http://dosser.anu.edu.au/index.php, a directory for "accessible"
equipment. You dont need to sign up to browse or search it, only to add new
equipment. Comments welcome!

cheers
Sally


Dr SJ Stowe
Facility Coordinator
ANU Electron Microscopy Unit
www.anu.edu.au/EMU/

On Mon, July 23, 2007 11:47 pm, stmccart-at-vt.edu wrote:
}

}
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} Hello: We are going to a web based sign up for a new characterization
} facility that will contain about 10 instruments. We have done a
} search of the archives and found 3 systems, FACES and one from
Northwestern and U.
} of Arizona. Are there others that people are using we should
} consider. Thanks. Steve
}
} Stephen McCartney
} Senior Research Associate
} Nanoscale Characterization and Fabrication Laboratory (NCFL) Institute
} for Critical Technology and Applied Science (ICTAS)
} 1991 Kraft Drive
} Va Tech
} Blacksburg, VA 24061
} 540-231-9765
}






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[Microscopy] web based instrument sign up, calendar,
18, 27 -- DOSSER for shared equipment
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==============================Original Headers==============================
31, 21 -- From syli-at-northwestern.edu Mon Jul 23 18:46:15 2007
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From: tina-at-pbrc.hawaii.edu
Date: Mon, 23 Jul 2007 21:54:43 -0500
Subject: [Microscopy] Parts available for S-800

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html

Hi, All-

We are taking apart a Hitachi S-800 FESEM for disposal. We have scavenged
some parts for another instrument, but some items are still available. If
you are interested in anything, please email me (tina-at-pbrc.hawaii.edu) as
soon as possible - it is coming apart within days!

Aloha,
Tina

****************************************************************************
* Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu *
* Biological Electron Microscope Facility * (808) 956-6251 *
* University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf*
****************************************************************************


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From: jacques.faerber-at-ipcms.u-strasbg.fr
Date: Tue, 24 Jul 2007 02:14:00 -0500
Subject: [Microscopy] Re: web based instrument sign up

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html

We use phpgroupware (http://www.phpgroupware.org/), for individual
calendar and instrument reservation.

It's not the most "clik and play" developed software, but it is
web-based (people work on mac, linux/unix and windows here), it's free
and open, and based on php, one can recover all the data for use for
statistics or billing.

We use it for all shared or service instruments (microscopes, x-ray
diffractor, AFM, etc.), but for meeting rooms or things like video
projector too.

It gives probably more achieved applications, but this one is enough for
our needs,... and budget !

J. Faerber
IPCMS-GSI
(Institut de Physique et Chimie des Matériaux de Strasbourg
Groupe Surface et Interfaces)
23, rue de Loess ; BP43
67034 Strasbourg CEDEX 2
France

Tel 00 33(0)3 88 10 71 01
Fax 00 33(0)3 88 10 72 48
E-mail Jacques.Faerber-at-ipcms.u-strasbg.fr



stmccart-at-vt.edu a écrit :
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} Hello: We are going to a web based sign up for a new characterization
} facility that will contain about 10 instruments. We have done a search of
} the archives and found 3 systems, FACES and one from Northwestern and U. of
} Arizona. Are there others that people are using we should
} consider. Thanks. Steve
}
} Stephen McCartney
} Senior Research Associate
} Nanoscale Characterization and Fabrication Laboratory (NCFL)
} Institute for Critical Technology and Applied Science (ICTAS)
} 1991 Kraft Drive
} Va Tech
} Blacksburg, VA 24061
} 540-231-9765
}
}
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From: mj_iqbal-at-yahoo.com
Date: Tue, 24 Jul 2007 07:45:58 -0500
Subject: [Microscopy] viaWWW: Bacteria in Root Nodule

Contents Retrieved from Microscopy Listserver Archives
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This Question/Comment was submitted to the Microscopy Listserver
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Email: mj_iqbal-at-yahoo.com
Name: Muhammad Javed

Title-Subject: [Filtered] Bacteria in Root Nodule

Question: Dear Randy
I am of the mind the suggestion given by Pat is of great worth but I
would like to add few more things .we are working on legume root
nodule from so long and results are the excellent even for the immunogold
labelling as well, where the section thickness, stability matters a
great. I am forwarding you the protocol (on and off list) along with the
tips that are common with a slight change. Discussion or details about
the topic might be long and other people would not have an interest in
that.
If you still face some problem or having quires donÇt hesitate to
write.
Muhammad Javed Iqbal
Electron Microscopist
Transmission Electron Microscopy Facility
TEM Labs
Ph # +92-41-651475-80
Ext #264
{mailto:mj_iqbal-at-yahoo.com} mj_iqbal-at-yahoo.com
{mailto:mjiqbal-at-nibge.org} mjiqbal-at-nibge.org
NIBGE Pakistan

Protocol for Ultra Structural Studies
Material:
Pipes Buffer [0.2M] (Piperzine-N,N-bis [2-ethanesulfonic acid]) FW :
302
Add 6gm Pipes to 50 ml of distilled water. Add 1N NaOH drop wise until
solid dissolves then adjust ph 6.8. Make up a volume of 100ml and store
at 4 0 C.
Fixative
Add 5 ml of (5 % v/v) glutaraldeyhde to 10 ml of 0.2M Pipes buffer.
Adjust ph 6.8.With 1N NaOH and make up volume to 25 ml.
Lead Citrate [4%] Pb(C6H5O7). 3H2O FW: 1053.82
Add 0.04 g Lead citrate salt to 10 ml of double distilled and
autoclaved water shake thoroughly then add 100 µl of 10 N NaOH solution.
Prepare fresh before use.
Make sure that Co2 present in the air should not trap with lead citrate
solution which may lead to precipitates.
Uranyl Acetatae [5%] UO2(CH3COO)2. 2H2O FW : 424.14
Dissolve 5 gm of Uranyl Acetate in 100 ml of distilled water (filter
sterilize with 0.2m Cellulose Acetate filter and store in cuvettes
covered with aluminum foil to save from direct light) used as stain to
enhance electron density.
Ethanol Series
30%, 50 %, 70 %, 100 % as dehydrating agent
Acetone
Absolute Acetone as transitional solvent
Spur Resin.
ERL / VCD 4206 (Vinylcyclohexene dioxide) 10 Parts
DER736 (diglycidyl ether of propylene glycol) 6
NSA (Nonyl succnic anhydride) 26
S1 (Dimethylaminoethanol) 0.4
Methods:
1.5% w/v Difco water agar was prepared after boiling in a microwave
oven and was kept in oven set at 37 0 C to prepare agar blocks. Small
pieces almost of 1mm X 1mm size from fresh root nodules were (cut under a
drop of fixative) were placed in 1.5% Difco agar that was poured in a
Petri dish and was left for 10 minutes at room temperature after agar
sets hard (solidifies). The agar (blocks) that were holding a thin layer
of agar around the nodule tissue was cut with a sharp scalpel Blade.
These blocks were immersed in 5%Glutaraldehyde prepared in 0.2 M Pipes
Buffer and Ph was set to 6.8. These small pieces were subjected to vacuum
in order to draw the fixative in to the tissue for half an hour.
Fixation took place in 18 Hrs placing the samples on a 55 0 fixed angle
specimen rotator at 5 rpm on room temperature. Rinsing of samples to remove
the fixative was performed in .2M Pipes buffer Ph 6.8 for three times
with an interval of 15 minutes each Post fixation was achieved
with 1% Osmium Tetroxide for 18Hrs at room temperature The tissue was
washed with autoclaved distilled water twice for 15 min and then
shifted to 5% Uranyl Acetate solution for 16-18 Hrs The tissue was again
washed with distilled waster twice for 15 min and then dehydrated with
Ethanol series comprising of 30%, 50 %, 70 %, 100% dilutions consequently
and kept in each fraction for 15 min and in 100 % thrice. 2 times for 1
hour and then before leaving overnight replace the ethanol with new
one. The tissue was left in absolute Acetone 2 X 30 min as transitional
solvent because the ethanol doesnÇt mixes well with spur Resin. Tissue
was infiltrated with a mixture of Spur embedding media. The ratio of
resin to acetone was as follows 1:3 , 1:2, 1:1 each for 8 hrs al least
and then absolute resin 100% initially for 8 hrs and then left overnight
.The samples were oriented in Moulds and resin cured at 60 0C for
48-60 Hrs.
The polymerized resin blocks were trimmed like a trapezoid and faced
with fine scalpel blade. Ultra thin serial sections of approx 120nm were
made with RMC MT 7000 ultra micro tome and placed on 300 mesh nickel
grid. Stained the grids with 5% Uranyl Acetate for 30 minutes and washed
three times with distilled water. Grids were double stained with Lead
Citrate for 10 minutes in NaOH chamber. Examination of sections was
performed with JEOL JEM1010 Transmission Electron Microscope operating at
80 kv available at TEM Facilities Lab National Institute for
Biotechnology & Genetic Engineering (NIBGE ) Faisalabad. Pakistan
Plastic sections are enough stable that you would not need help of any
plastic or carbon film
Note: For mixing and infiltration the tissue was kept on specimen
rotator
All the steps were adopted at room temperature almost 24-26 0C



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From: kraftpiano-at-gmail.com
Date: Tue, 24 Jul 2007 11:26:33 -0500
Subject: [Microscopy] SEM- Vacuum pump is bound.

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html

Does anybody have a tip on how to un-stick a vacuum pump? I have a
JEOL JSM-35C that I'm getting up and running, and one of the pumps is
bound. I can turn it by hand, but it takes a lot of effort. The
motor spins, and is right now grinding away at the belt when it turns,
since it lacks the torque necessary to turn the vacuum pump wheel.

What could cause it to bind up like that? It has enough oil in it,
and the oil looks good.

--Justin.

==============================Original Headers==============================
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3, 26 -- Subject: SEM- Vacuum pump is bound.
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From: NWWhite-at-bwxt.com
Date: Tue, 24 Jul 2007 12:00:16 -0500
Subject: [Microscopy] SEM- Vacuum pump is bound.

Contents Retrieved from Microscopy Listserver Archives
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Justin,

Assuming a rotary, vane type roughing pump...

Could be the seals or bearings, but it has been my experience that most
often the sliding vanes and guides are damaged and/or sticking. I have
repaired this type of problem, but it requires pump disassembly,
cleaning, and parts replacement. Usually repair entails replacement of
the vanes, guide pins and vane springs. It would be good to replace the
shaft oil seals while you are at it.

If you can find a service manual for the pump it would be most helpful.
For some pumps, it is not immediately obvious what to remove for
disassembly. Furthermore, some require eccentric bearings be rotated to
achieve the proper vane to housing clearances.

Another possibility... If the torque requirement is within specification
(even though stiff) and the drive motor is a capacitor start type, a bad
starting capacitor will cause failure to start. A bad capacitor will
reduce the motor start torque to near zero.

Woody



NW (Woody) White Jr
BWXT Services
Lynchburg, VA
http://www.bwxt.com/operations/semlab.html


-----Original Message-----
X-from: kraftpiano-at-gmail.com [mailto:kraftpiano-at-gmail.com]
Sent: Tuesday, July 24, 2007 12:27 PM
To: White, Woody N.

Does anybody have a tip on how to un-stick a vacuum pump? I have a
JEOL JSM-35C that I'm getting up and running, and one of the pumps is
bound. I can turn it by hand, but it takes a lot of effort. The
motor spins, and is right now grinding away at the belt when it turns,
since it lacks the torque necessary to turn the vacuum pump wheel.

What could cause it to bind up like that? It has enough oil in it,
and the oil looks good.

--Justin.

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From: gma27824-at-csun.edu
Date: Tue, 24 Jul 2007 17:31:27 -0500
Subject: [Microscopy] AskAMicroscopist: TEM of Arthrobacter GFB100

Contents Retrieved from Microscopy Listserver Archives
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Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (gma27824-at-csun.edu) from http://microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Tuesday, July 24, 2007 at 13:31:24
---------------------------------------------------------------------------

Email: gma27824-at-csun.edu
Name: Guadalupe Aguirre

Organization: California State University, Northridge (CSUN)

Education: Graduate College

Location: Northridge, CA, USA

Question: I am about to be trained to use a TEM to take electronmicrographs of a bacterium called Arthrobacter GFB100 that might be producing a biofilm. As very few people work with it, I am looking at a protocol performed on a similar genus. I have two questions: 1)In the fixation step, OsO4 in Veronal-sodium acetate is used. What is veronal-sodium acetate, and why would it be used as opposed to more conventional fixation agents? 2)After polymerization, the sample was freed from a glass coverslip by shock-freezing in liquid nitrogen. Is this absolutely necessary or are there other alternatives? The paper was published by the Journal of Bacteriology, Jan. 1997, p.31-40, vol.179, No.1, entitled, "Metal Selectivity of In Situ Microcolonies in Biofilms of the Elbe River"; the authors are H. Lundsdorf, I. Brummer, K. N. Timmis, and I. Wagner-Dobler. Any information would be much appreciated-Thank you!

---------------------------------------------------------------------------

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From: kraftpiano-at-gmail.com
Date: Tue, 24 Jul 2007 22:14:50 -0500
Subject: [Microscopy] Kevex Delta-3 EDS question

Contents Retrieved from Microscopy Listserver Archives
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Does anybody have the pinouts and connection diagrams for the Kevex Delta-3
Xray analyzer? We received the full Kevex system with our donated JSM-35C,
and I would like to be able to use it for imaging, but the cables were not
included.

Oh, and for those who remember my employment situation, I've landed a much
better position teaching forensics, physics, chemistry and astronomy at
another school, still to high school. And the new school is much better
about letting me be a little, um, non-traditional with my teaching methods.
(More labs, less book.)

--Justin A. Kraft
Leadership Academy
West Palm Beach, FL.

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From: gary-at-gaugler.com
Date: Tue, 24 Jul 2007 23:47:02 -0500
Subject: [Microscopy] Re: Kevex Delta-3 EDS question

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html

Excuse me, but I think you need to spend some
serious time working with the SEM system.
Whatever it is. You had a Jeol system then an
ISI system and now problems with EDS.

This is a bit too much, IMO. Free stuff has a
price. There are no free lunches here.

Reality is a shocking wall of resistance.
I do not mean to be combative or negative.

However, you need to be, IMO, more realistic
in the scope of what you are trying to do.
It is not trivial by any measure. But if
you can prevail---all the better for you.

Stand back and prioritize your issues. If
no beam, EDS is irrelevant. Get the SEM
stable and then move forward.....my opinion.

Do you have a beam of quality???

gary g.


At 07:16 PM 7/24/2007, you wrote:




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From: jb_sanderson-at-yahoo.com
Date: Wed, 25 Jul 2007 14:25:26 -0500
Subject: [Microscopy] Re: Are you there

Contents Retrieved from Microscopy Listserver Archives
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Dear David,

Yes. It collects two microscopy listservs. I rarely
look at it; your last incoming message was Feb '06.
Contact via Exeter Road is best.

Gum

--- David Sanderson {David-at-SandersonsGarage.co.uk}
wrote:

} Hi Gum,
}
} Are you still on this e-mail address.
}
} David
}



___________________________________________________________
Yahoo! Answers - Got a question? Someone out there knows the answer. Try it
now.
http://uk.answers.yahoo.com/

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From: bozzola-at-siu.edu
Date: Wed, 25 Jul 2007 15:16:27 -0500
Subject: [Microscopy] Re: AskAMicroscopist: TEM of Arthrobacter GFB100

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html

Veronal - sodium acetate is a buffer composed of sodium barbitol and
sodium acetate. It was made popular by Ryter & Kellenberger many
decades ago to preserve "bacterial plasms." Even now, I use it
sometimes to fix stubborn organisms. However, it is a controlled
substance (a barbiturate) and probably difficult to obtain without a
license. With hard to fix organisms, I often fix overnight (at room
temperature) in osmium tetroxide buffered in veronal acetate.

You should try a more conventional buffer (one of the phosphates, for
example) for various times. Try following the protocol that you
mention only use phosphate buffer and extend the times a bit.

The reason for LN2 shocking was to release the organisms. Unless
there is a good reason for culturing on glass substrates, use a
plastic petri dish instead (or a coverglass made of plastic or
Aclar), and do a monolayer embedding. That way, you can examine the
cells as they grow (in situ) on the plastic surface. I have a
technique for doing this. If you are interested, I can dig it out.

JB

} Question: I am about to be trained to use a TEM to take
} electronmicrographs of a bacterium called Arthrobacter GFB100 that
} might be producing a biofilm. As very few people work with it, I am
} looking at a protocol performed on a similar genus. I have two
} questions: 1)In the fixation step, OsO4 in Veronal-sodium acetate is
} used. What is veronal-sodium acetate, and why would it be used as
} opposed to more conventional fixation agents? 2)After
} polymerization, the sample was freed from a glass coverslip by
} shock-freezing in liquid nitrogen. Is this absolutely necessary or
} are there other alternatives? The paper was published by the Journal
} of Bacteriology, Jan. 1997, p.31-40, vol.179, No.1, entitled, "Metal
} Selectivity of In Situ Microcolonies in Biofilms of the Elbe River";
} the authors are H. Lundsdorf, I. Brummer, K. N. Timmis, and I.
} Wagner-Dobler. Any information would be much appreciated-Thank you!


--
+++++++++++++++++++++++++++++++++++++++++++++++++++++++

John J. Bozzola, Ph.D., Director
Integrated Microscopy & Graphics Expertise (IMAGE)
Southern Illinois University
750 Communications Drive - MC 4402
Carbondale, IL 62901
Telephone: 618-453-3730

+++++++++++++++++++++++++++++++++++++++++++++++++++++++

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From: jmkrupp-at-ucsc.edu
Date: Wed, 25 Jul 2007 15:40:18 -0500
Subject: [Microscopy] TEM after formaldehyde?

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html

Hi again:

Just checking before I try something new.

Researcher gave me a sample of diatoms in 4% formaldehyde in sea
water. She made a standard collection and used her usual formaldehyde
fix since it was going to be for light microscopy and population
monitoring only.

Well, some weird things showed up, now she wants to check them using
TEM to see if there are any viruses inside or if she can figure out
what else might be going on.

I, of course, have been called on to do the EM. So, I'm trying to
work out something that makes sense. The guys are in 4% formaldehyde
in sea water. Wondering about rinsing out SW before or after OsO4 and
or choice of buffer, if needed.

We know these guys may have suffered from the formaldehyde and may
not turn out so well, but figure its worth a try. Any hints?

Thanks

Jon
--

Jonathan Krupp
Microscopy & Imaging Lab
C230 Earth & Marine Science
University of California
Santa Cruz, CA 95064
(831) 459-2477
jmkrupp-at-ucsc.edu

I rode in AIDS/Lifecycle 6 SF to LA June 3 - 9, 2007 to raise money
for the San Francisco AIDS Foundation. Visit
http://www.aidslifecycle.org for more information about the ride or
http://www.aidslifecycle.org/5482 to make a donation.

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From: gabriel.corfas-at-childrens.harvard.edu
Date: Wed, 25 Jul 2007 15:45:41 -0500
Subject: [Microscopy] Control board for Jung 2800 E cryostat

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html

We have a Jung 2800 E cryostat that apparently does not work due to a
problem in the main control electrical board.

Leica informed us that they do not have replacement parts and they do not
repair the board.

Does anybody know of a source of replacement boards or of somebody who
repairs them?

Thanks
GC

--
Gabriel Corfas, Ph.D.
Neurobiology Program
Children's Hospital
300 Longwood Avenue
Boston, MA 02115

Tel: 617-355-7364
Fax: 617-730-0242



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From: bryan_hansen-at-earthlink.net
Date: Wed, 25 Jul 2007 17:13:06 -0500
Subject: [Microscopy] Platinum Blue stain

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html

Hi,

I read a paper recently on using platinum blue as an alternative to Uranyl Acetate for en bloc and post section staining. Does anybody have the recipe for how to make it? I contacted one of the writers of the paper and the recipe I have is in japanese so if anyone has an english version I would greatly appreciate it.

Thanks,
Bryan Hansen
Rocky Mountain Laboratory

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From: connellyps-at-nhlbi.nih.gov
Date: Wed, 25 Jul 2007 18:38:33 -0500
Subject: [Microscopy] Re: TEM after formaldehyde?

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html

Jon,

Are you sure there is biological material inside the crusty diatoms?

I had done some work with diatoms by putting them whole onto coated grids
but that was for identification, not to see the insides. I did get some
lovely pictures and found one that was not known to be present in North
America!
We did not have a SEM back then - 1972 I think.

You can do a 1% glutaraldehyde fix in sea water (did that for marine animal
sperm) then go into buffer (I used phosphate) to wash and osmium fix.

I would not wish to thin section these critters without some advice from
someone who had done so successfully. I had a lot of trouble with sponges
many years ago. The spicules(?) made a mess of the sections. This was
before I joined the Listserv and did not know anyone to ask for help.

Try glass knives before diamond I would think.

Lost of luck,
Pat

Patricia Stranen Connelly
Lead Technologist
NHLBI Electron Microscopy Core
National Institutes of Health
14 Service Road South
Bldg. 14E ­ Rm. 111B MSC 5570
Bethesda, MD 20892-5570
Phone 301-496-3491
connellyps-at-mail.nih.gov


On 7/25/07 4:44 PM, "jmkrupp-at-ucsc.edu" {jmkrupp-at-ucsc.edu} wrote:
} Just checking before I try something new.
}
} Researcher gave me a sample of diatoms in 4% formaldehyde in sea
} water. She made a standard collection and used her usual formaldehyde
} fix since it was going to be for light microscopy and population
} monitoring only.
}
} Well, some weird things showed up, now she wants to check them using
} TEM to see if there are any viruses inside or if she can figure out
} what else might be going on.
}
} I, of course, have been called on to do the EM. So, I'm trying to
} work out something that makes sense. The guys are in 4% formaldehyde
} in sea water. Wondering about rinsing out SW before or after OsO4 and
} or choice of buffer, if needed.
}
} We know these guys may have suffered from the formaldehyde and may
} not turn out so well, but figure its worth a try. Any hints?
}
} Thanks
} Jon



==============================Original Headers==============================
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12, 24 -- Subject: Re: [Microscopy] TEM after formaldehyde?
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From: opmills-at-mtu.edu
Date: Wed, 25 Jul 2007 19:33:01 -0500
Subject: [Microscopy] viaWWW: lab climate control

Contents Retrieved from Microscopy Listserver Archives
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using the WWW based Form at http://www.microscopy.org/MicroscopyListserver/MLFormMail.html
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Email: opmills-at-mtu.edu
Name: Owen Mills

Organization: Michigan Tech Univ

Title-Subject: [Filtered] lab climate control

Question: All,

I'm curious whether any of you are using the rolling air conditioning units with a vent hose. If so, do they work? What size units are you using (in BTU's).

Thanks

Owen Mills
Michigan Tech University


---------------------------------------------------------------------------

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From: nizets2-at-yahoo.com
Date: Thu, 26 Jul 2007 06:15:20 -0500
Subject: [Microscopy] AskAMicroscopist: TEM of Arthrobacter GFB100

Contents Retrieved from Microscopy Listserver Archives
http://www.microscopy.com/MicroscopyListserver/MicroscopyArchives.html

Hi!

I also recommended (off list) flat embedding, however
definitely not in a plastic dish! The reason is that
the plastic dish is not perfectly flat, and when you
flat embed the surface of the resin bloc is full of
holes. This means you'll need 8-10 100 nm thick
sections before obtaining a perfectly flat surface.
This is probably the thickness where the bacteria are
to be found!
Of course you can still try to image your bacteria in
sections full of holes, this is another story.

An alternative would be to actually grow on glass
coverslips and flat embed in eppendorf tubes whose
bottom has been cut out. The glass coverslips offer a
perfectly flat surface, and if you approach the knife
(well actually the spectimen) carefully enough, the
first few sections may be ok.
Other more expensive alternative exist (thermanox
coverslips for example).

Best regards,

Stephane


--- bozzola-at-siu.edu wrote:

}
}
}
}
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}
} Veronal - sodium acetate is a buffer composed of
} sodium barbitol and
} sodium acetate. It was made popular by Ryter &
} Kellenberger many
} decades ago to preserve "bacterial plasms." Even
} now, I use it
} sometimes to fix stubborn organisms. However, it is
} a controlled
} substance (a barbiturate) and probably difficult to
} obtain without a
} license. With hard to fix organisms, I often fix
} overnight (at room
} temperature) in osmium tetroxide buffered in veronal
} acetate.
}
} You should try a more conventional buffer (one of
} the phosphates, for
} example) for various times. Try following the
} protocol that you
} mention only use phosphate buffer and extend the
} times a bit.
}
} The reason for LN2 shocking was to release the
} organisms. Unless
} there is a good reason for culturing on glass
} substrates, use a
} plastic petri dish instead (or a coverglass made of
} plastic or
} Aclar), and do a monolayer embedding. That way, you
} can examine the
} cells as they grow (in situ) on the plastic surface.
} I have a
} technique for doing this. If you are interested, I
} can dig it out.
}
} JB
}
} } Question: I am about to be trained to use a TEM to
} take
} } electronmicrographs of a bacterium called
} Arthrobacter GFB100 that
} } might be producing a biofilm. As very few people
} work with it, I am
} } looking at a protocol performed on a similar genus.
} I have two
} } questions: 1)In the fixation step, OsO4 in
} Veronal-sodium acetate is
} } used. What is veronal-sodium acetate, and why would
} it be used as
} } opposed to more conventional fixation agents?
} 2)After
} } polymerization, the sample was freed from a glass
} coverslip by
} } shock-freezing in liquid nitrogen. Is this
} absolutely necessary or
} } are there other alternatives? The paper was
} published by the Journal
} } of Bacteriology, Jan. 1997, p.31-40, vol.179, No.1,
} entitled, "Metal
} } Selectivity of In Situ Microcolonies in Biofilms of
} the Elbe River";
} } the authors are H. Lundsdorf, I. Brummer, K. N.
} Timmis, and I.
} } Wagner-Dobler. Any information would be much
} appreciated-Thank you!
}
}
} --
}
+++++++++++++++++++++++++++++++++++++++++++++++++++++++
}
} John J. Bozzola, Ph.D., Director
} Integrated Microscopy & Graphics Expertise (IMAGE)
} Southern Illinois University
} 750 Communications Drive - MC 4402
} Carbondale, IL 62901
} Telephone: 618-453-3730
}
}
+++++++++++++++++++++++++++++++++++++++++++++++++++++++
}
} ==============================Original
} Headers==============================
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} 2007
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} 9, 17 -- From: "John J. Bozzola" {bozzola-at-siu.edu}
} 9, 17 -- Subject: Re: [Microscopy] AskAMicroscopist:
} TEM of Arthrobacter GFB100
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____________________________________________________________________________________
Pinpoint customers who are looking for what you sell.
http://searchmarketing.yahoo.com/

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From: nizets2-at-yahoo.com
Date: Thu, 26 Jul 2007 09:32:02 -0500
Subject: [Microscopy] Glue without carbon (SEM)

Contents Retrieved from Microscopy Listserver Archives
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Hi all!

I am trying to detect organic contamination on/in
aluminosilicate material by SEM+ED