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From: zaluzec-at-microscopy.com
Date: Sun, 1 Jan 2006 17:52:42 -0600
Subject: [Microscopy] Microscopy Listserver2005 Archives Now On-Line

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Happy New Year Colleagues:

The complete 2005 Archives are now on-line at:

http://www.microscopy.com


Cheers..

Nestor
Your Friendly Neighborhood SysOp

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From: Colin.Veitch-at-csiro.au
Date: Mon, 2 Jan 2006 18:38:14 -0600
Subject: [Microscopy] Restarting a JEOL 2010 on ion pumps

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G'day and happy new year,

Over the Christmas break we routinely shut our microscopes down to run
only on ion pumps as our cooling water can be a little "temperamental".


This year we had a power glitch which turned the microscopes completely
off. To top it off our cooling water is out of action for maintenance
until next week!! I was hoping to get all the ion pumps running on the
microscopes to keep the vacuums at leat a bit clean and I have managed
to get this done on all but our JEOL 2010. Does anyone know if it is
possible to get just the ion pumps running without needing cooling water
on a 2010?

Any help would be greatly appreciated.

Thank you very much.

Colin Veitch

Electron Microscopist

CSIRO Textile and Fibre Technology

PO Box 21, BELMONT, Vic. 3216. Australia.

E-mail: colin.veitch-at-csiro.au

Web: http://www.tft.csiro.au

Tel: +61 (0) 3 5246 4000
Mob: 0438 538 475
Fax: +61 (0) 3 5246 4811



The information contained in this e-mail message may be privileged or
confidential information. If you are not an intended recipient, you may
not copy, distribute or take any action in reliance on it. If you have
received this message in error, please telephone CSIRO Textile and Fibre
Technology on +61 3 5246 4000.



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From: brakenho-at-science.uva.nl
Date: Tue, 3 Jan 2006 02:18:51 -0600
Subject: [Microscopy] FOM2006 abstract deadline 9 Jan. near! FocusOnMicroscopy, Perth,

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FOCUS ON MICROSCOPY 2006, Perth, Australia, April 9-12, 2006
19th International Conference on 3D Image Processing in Microscopy 18th
International Conference on Confocal Microscopy

Dear Colleagues

January 9, 2006, the deadline for abstract submission for the Perth
conference is nearing.

Please submit your abstract by that date.

The program for the conference will be finalized and available on our
website on Jan 23, 2006. Authors will also be informed individually by
E-mail on the placement of their contribution in the program.
Abstracts for oral and poster presentations are sollicited. Submission
preferably through the conference website:
http://focusonmicroscopy.org/
where also the conference registration is open and hotel booking
information is available.

The earlier than usual deadline of 9 Jan. was chosen to give delegates
sufficient time -after the program has been announced- to arrange their
travel and acccommodation before the conference starting date of Sunday 9
April, 2006.

Please note that the conference will take place in the Esplanade Hotel in
Fremantle, the historic waterfront suburb of Perth, See further our
website.

The program will start on Sunday April 9, around 18 hours with an
opening symposium in the Esplanade hotel followed by a welcome reception.

The conference Focuson Microscopy 2006 will take place as the next in a
series of unique interdisciplinary meetings on advanced multidimensional
light microscopy and image processing. The conference will be hosted by
the University of Western Australia in Perth.

Focus on Microscopy 2006 is the continuation of a conference series
presenting the latest innovations in optical microscopy and its
applications in biology, medicine, material science, and information
storage. 3D optical imaging and related theory are traditional subjects
for the conference. The conference series is also known for covering the
rapid development of advanced fluorescence labeling techniques for the
confocal and multi-photon 3D imaging of -live- biological specimens.
This year, in addition, special attention will be given to imaging in
thick tissues.

Abstracts for contributions are invited and can now be submitted through
the website:

www.FocusOnMicroscopy.org

where further information on the present and previous FOM conferences can
be found.

Important dates:

Deadline for the submission of abstracts: January 9, 2006
Draft program available on the web: January 23, 2006
at website www.FocusOnMicroscopy.org
Deadline for early registration: February 20, 2006

Welcoming you to beautiful Perth for the FOM2006
conference and exhibition.

With the best wishes for the New Year and on behalf of the organizing
committee,

David Sampson,
University of Western Australia, Perth, Australia

Fred Brakenhoff
Swammerdam Institute for Life Sciences, University of Amsterdam, The
Netherlands

E-mail: info2006-at-FocusOnMicroscopy.org

Web: www.FocusOnMicroscopy.org

















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From: fisiltd-at-gmail.com
Date: Tue, 3 Jan 2006 08:39:50 -0600
Subject: [Microscopy] viaWWW: O'Brien & McCully, Study of Plant Structure

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Email: fisiltd-at-gmail.com
Name: Howard Freeman

Organization: FISI Ltd

Title-Subject: [Filtered] O'Brien & McCully, Study of Plant Structure

Question: I have a copy of The study of Plant Structure, O'Brien & McCully, first printing, 1981 in very good condition. I used this for my teaching but have moved away from histology.
If anyone is interested please send email to fisiltd-at-gmail.com

Thanks

H Freeman

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From: soraia.pirfo-barroso-at-insa-lyon.fr
Date: Tue, 3 Jan 2006 08:40:27 -0600
Subject: [Microscopy] viaWWW: LM on 304L grey superficial layer and black lines

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Email: soraia.pirfo-barroso-at-insa-lyon.fr
Name: Soraia Pirfo Barroso

Organization: pos-doc at INSA - Institut National des Sciences AppliquČes de Lyon

Title-Subject: [Filtered] LM on 304L grey superficial layer and black lines

Question: I have shot peening 304L steel specimens, thickness about 5mm. Objective to visualise martensitic phase.
I used tradional mechanical polishing, till 1200 grade, different etching based on water, HCl and potassium dissulfite (Berahas's tint).
I did not clearly observe a martensitic layer but some insland, there is a dark grey to black layer on treated surface of about 10 microns, in case of highest resolution 200x I see it bright grey. What you believe it to be?
I observe black flowing lines, variable lengths from 5 to 200 microns , paralel to surface through all specimen thickness and all its length. What it cab be? Some mention they are mechanical polishing effect others ferrite phase.

Thanks for whom took time reading or answering!
Best new year wishes!
Soraia Pirfo

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From: mike.reedy-at-cellbio.duke.edu
Date: Tue, 3 Jan 2006 11:06:54 -0600
Subject: [Microscopy] Re: viaWWW: Potassium Permanganate

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Hi Sue,
You give me an occasion to offer via the
listserver some lore on MnO4 staining and epoxy
resin adaptations to MnO4 staining that I
provided off-listserver earlier this year in 2
other messages. I'll offer my message in three
lengths.

SHORT VERSION is: Using epoxy embedding that
excludes MNA (MNA?), MnO4 section staining
followed by a lead stain (Sato's) can give
fantastic contrast to everything --- not just
membranes --- including any dirt/oil collected
from the trough when picking up the sections, or
rinsed off the pipette tip when dispensing drops
of stain. The contrast is even greater if tannic
acid has been used in the fix. Section staining
of Epon-DDSA or Spurr embeddings is OK, but
Araldite gives distinctly finer-grain
higher-resolution staining.


MEDIUM VERSION:
NMA renders use of KMnO4 as a section stain
impossible; MnO4 reacts with the cured resin.
Reedy, J Cell Biol 1965 26:309-11

We prefer the Mn-Pb sequence for higher contrast
generally, and especially in the thinnest
sections - 15-30 nm. So we gave up NMA about 30
years ago, and just heretically fiddled the
resin-DDSA proportions until we got a hardness we
liked; it worked just fine. With Epon that was
that. We prefer Araldite because it is less even
reactive than Epon or Spurr with MnO4. With
Araldite-DDSA, we use Araldite 506. We found the
best mixture still a bit too brittle, so added a
bit of DER 736 as a "plasticizer", have used that
for over 30 years.
Araldite 506 50g 10g
DDSA 75g 15g
DER 736 10g 2g
1.8% v/v DMP-30 (range 1.5% to 2.0%).
Reference Reedy et al (1983) J. Muscle Res. Cell Motil. 4:25-53.


LONG LONG VERSION (data dump):
(This comes mostly from my off-listserver reply
to some contributors to NetNotes in the May or
June 2005 Microscopy today, regarding a query by
Ursula J Potter about MnO4 use as a section
stain.)

I have used 1-2% MnO4 section staining since
1964, almost exclusively since 1970, always
followed by Pal's bleach (see below) to eliminate
surface "pepper" of MnO2 ppt, followed then by a
lead stain, of which Sato's appeals to us as the
best and most storage-stable (Hanaichi et al,
1986, Proc 11th Internat'l EM Congress (Kyoto), p
2181). After publishing my brief note on the MNA
problem (J Cell Biol, 1965, 26:309), I soon
switched completely to Araldite-DDSA embedding,
because it turned out that cured Araldite was
much less reactive with MnO4 than any other epoxy
resin mix I ever tested, including Spurr's when
it later appeared. I used an extreme TEST,
exposing cured blocks of various embedding resin
mixtures to HOT KMnO4 solution (test tube
immersed in boiling water) for 1 hr. In that
test, the Epon-DDSA resin "preferred" in my Brief
Note developed a blackened micro-fissured
tree-bark surface (like that of MNA-Epon in
unheated KMnO4, while the Araldite-DDSA resin was
scarcely discolored at all. The Epon-DDSA
sections typically presented a more nano-granular
staining than did Araldite sections.

I think I gave up using barium MnO4 because it
seemed to give blotchy staining more often than
KMnO4. I also guessed that barium ion might
capture carbonate as Pb did; Pb carbonate was the
ubiquitous contaminant from hell with all the
early lead stains that preceded lead citrate.

We wanted the maximum staining contrast that is
provided by the Mn-Pb sequence because we mainly
use 15-30 nm sections. Adding in UrAc to the
sequence helps not at all. We do use UrAc as a
block-stain, or as a secondary post-fixative
(preferred to OsO4 in my lab) when our primary
fixative is tannic acid with or (usually) without
glutaraldehyde (TA-URAC works beautifully on
myofilaments and on residual membranes in our
permeabilized demembranated insect muscle, both
in aqueous buffers at room temperature and as
freeze-subst'n chemistry in acetone at -80 to
-90°C). Recently, we have even found that the
actin monomer substructure of thin myofilaments
can be seen-- it appears to be relatively
negatively stained, coated with metal derived
from the TAURAC and Mn-Pb sequences (Fig 3, blue
outlined: Fig 5 B; Liu et al, J Struct Biol,
2004, 147:268-282).

We had to experiment to find the optimum staining
times for KMnO4, Pal's bleach washing, and lead
stain needed to saturate contrast without
"digesting" or extracting structures out of the
sections. We learned by deliberately
over-treating sections with each step. 60-90 min
will remove Z-bands and (maybe) myofilaments from
100-150 nm Araldite sections, leaving a negative
image, actually a cavity in the resin. Staining
saturatees slower in Araldite than in Epon
sections. 100 nm sections become stain-saturated
slower than 25 nm sections. Cross-sections of
muscle saturate much faster than longitudinal
sections. 2-10 min in MnO4, 10-30s washing in
Pal's bleach 1:100, and 1-5 min on Sato's Pb
stain works for 25 nm longitudinal sections of
muscle.

Stain apparently penetrates the section only
where the pure embedding medium is modified by
the presence of stained structures that intercept
the section surface. The stains percolate only
through the embedded structures, NOT through
blank empty resin regions, as we found when
sections had piled up or folded two-deep on one
another; the covered section (sandwiched between
carbon support film and an overlying surface
section) would appear stained only within 0.1-0.2
um laterally from wherever the overlying section
contained embedded structures such as filaments
or membranes to conduct stain from the solution
to the underlying structures in a covered section.

PAL's BLEACH,
from the J. D. Robertson 1963 paper cited in my 1965 brief note:
Distilled water 100 ml
Potassium sulphite 0.5 g
Oxalic acid 0.5 g
dilute 1:100 for use (3-4 drops of above stock
in 10 ml water), and run 10-20 drops over the
grid after MnO4 staining. Water wash immediately
after MnO4 and after Pal's.

All our staining is done by floating grids on
small spherical drops of stain on fresh-washed
Parafilm. (I'm not familiar with the
vertical-insertion immersion staining of grids
mentioned by Dr W. Muss in his helpful reply to
you Dec 30.) Keys to clean staining: draw up the
stains from under the surface in the stock
bottles, discard the first 3-6 drops before
putting the drop to be used down on a freshly
washed Parafilm surface. Mistrust paranoically
all unwashed surfaces on forceps and parafilm and
pipette barrels as likely to have finger grease
that will get on your sections or support films
and become intensely stained contaminants. Wash
everything you mistrust that will contact
sections and grids with lavish flows and jets of
deionized or distilled water. Stock lead stain
needs replacing when it starts to "fails" in
producing contrast, as often as every 4 weeks.
Stock 2% KMnO4 lasts "forever"; just remember to
respect and go below its MnO2-dirty surface when
filling your pipette.


Around 1970 I switched to a lower-viscosity
Araldite 506 mix (with DDSA, DER 732, DMP-30: see
MEDIUM VERSION above for recipe and reference)
that we have used ever since. It becomes as
water-thin in viscosity as Spurr's when heated to
60°C, so we rotate vials on a small tilted
platform in the oven for 20' x 3 changes to
optimize infiltration, bypassing the 50:50 (etc)
intermediate solvent-resin mixtures to go
directly from solvent into 100% accelerated resin
mix. We soon abandoned some of the other old-time
religion as well, giving up propylene oxide
completely to go directly from acetone or
ethanol, after we tried deliberately curing 10-15
ml vials of accelerated resin with 10-20% solvent
thoroughly blended in; the PO additive produced a
much "cheesier" final cured resin than equivalent
percentage of acetone or EtOH. (However, see my
Dec 6 2005 on-listserver message RE: LR White
contrast, calling attention to SW Brorson's use
of propylene oxide and/or high [accelerator] to
improve exposure of antigens in epoxy sections to
immunolabeling). We also explored the
epoxy-anhydride ratio over a wide and quite
heretical range (30-70% v/v anhydride) to finally
settle on a "nicer" final cured resin (less
brittle during block-trimming than the gospel
ratio) because we found acceptable cutting,
staining, and structural preservation over the
whole range we tried.

Some issues I consider still unfinished:
1) Can shorter or colder (etc) staining of
Epon(replacement)-DDSA or Spurr sections
eliminate the nano-granularity to approach the
finer-grained stain we routinely get in Araldite?
2) Does our Mn-Pb section stain perhaps "clog"
or "fill in" some fine structures in the 5 nm and
smaller range that are more delicately and
clearly brought out by the Ur-Pb staining
sequence? I've yet to re-check my impression
that the 4.8 nm axial repeat on thick filaments
routinely showed up 30 years ago on optical
diffraction of insect muscle section EMs, before
we started using the MnPb exclusively; and that
it never shows up now that we no longer use the
weaker contrasting UrPb sequence.

-mike reedy-


At 9:28 PM -0600 12/29/05, susan.vanhorn-at-stonybrook.edu wrote:
} ----------------------------------------------------------------------------
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--
-mike reedy-

************************
Michael K. Reedy, M.D.
Duke Univ. Med. Center
Dept. Cell Biology, Box 3011 (for U.S. Mail)
458 Alex Sands Bldg, Research Dr (courier)
Durham, NC 27710

Office 919-668-2534
Lab 919-684-5674
Fax 919-681-9929
mike.reedy-at-cellbio.duke.edu
http://note.cellbio.duke.edu/Faculty/Research/Reedy.html


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From: ramadanhany-at-gmail.com
Date: Tue, 3 Jan 2006 18:28:13 -0600
Subject: [Microscopy] Re: Tantalum polishing

Contents Retrieved from Microscopy Listserver Archives
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The easiest way to get a transmitted light image on the 510 is to just click
on ChD, which is a transmitted light detector. This will collect all the
light that passes through your sample AT THE SAME TIME you are collecting
confocal fluorescent images. So no need to double-expose your sample. You
will still need to set the condenser and neutral density filters, etc., for
Koehler.

I am not sure if it is a bug that the HAL light goes off when scanning - I
think it is probably a protection (safety) so the PMTs do not get blasted.

-Holly
__________________
Holly L. Aaron
CRL Molecular Imaging Center
http://imaging.berkeley.edu

3rd Annual Advanced Optical Microscopy Workshop
http://imaging.berkeley.edu/FLIM.html

-----Original Message-----
X-from: lubo-at-berkeley.edu [mailto:lubo-at-berkeley.edu]
Sent: Wednesday, December 28, 2005 6:46 AM
To: hollya-at-berkeley.edu

Thanks guys, I really apreciate it. I will try some of the mentioned
methods and I will update you with the results and I will tell you
which method(s) worked with me.

Best Regards

Hany

On 12/30/05, ramadanhany-at-gmail.com {ramadanhany-at-gmail.com} wrote:
}
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} Good morning,
}
} I grow tantalum oxide electrochemically on tantalum substrates, the
} issue is that using SEM I can say that the surface of tantalum oxide
} is full of defects "pin holes". I just use mechanical polishing of
} tantalum before growing oxide, so I think that these defects result
} from impurities of polishing sand papers. Is there any other way I can
} polish tantalum to the finest grade avoiding these impurities?
}
} I appreciate your responses.
}
} Thanks
} --
} **********************************************************
} Hany Ramadan
} Graduate student
} Chemistry department
} McMaster university, Hamilton, Ontario, Canada
} 905-525-9140 x: 26322
} elsayeh-at-mcmaster.ca
} **********************************************************
}
}
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--
**********************************************************
Hany Ramadan
Graduate student
Chemistry department
McMaster university, Hamilton, Ontario, Canada
905-525-9140 x: 26322
elsayeh-at-mcmaster.ca
**********************************************************


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From: CGoldsmith-at-cdc.gov
Date: Tue, 3 Jan 2006 20:10:02 -0600
Subject: [Microscopy] viaWWW: SEMS annual meeting for 2006

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Email: CGoldsmith-at-cdc.gov
Name: Cynthia Goldsmith

Organization: SEMS and CDC

Title-Subject: [Filtered] SEMS annual meeting for 2006

Question: This year the annual meeting for the Southeastern Microscopy Society (SEMS) will be held jointly with the Association of Southeastern Biologists (ASB). The meeting will be March 29-April 1 in Gatlinburg, Tennessee at the Gatlinburg Convention Center. This event brings together approximately 800 biologists from across the southeastern United States. Interests are diverse, but range from genetics and molecular biology, to physiology and population biology, to community and ecosystem ecology.

Registration deadline is January 17, 2006 and information is available at the SEMS website:

http://www.semicroscopy.org/index.html
or
http://www.semicroscopy.org/meetings/call-for-papers.html


Cynthia S. Goldsmith
Secretary, Southeastern Microscopy Society (SEMS)
Mailstop G32
Centers for Disease Control and Prevention (CDC)
Atlanta, GA 30333


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From: HWitkiewicz-at-SKCC.org
Date: Tue, 3 Jan 2006 20:10:42 -0600
Subject: [Microscopy] viaWWW: Comparing cryo- and cryo-immuno diamond knives

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Email: HWitkiewicz-at-SKCC.org
Name: Halina Witkiewicz

Organization: Sydney Kimmel Cancer Center

Title-Subject: [Filtered] Comparing cryo- and cryo-immuno diamond knives

Question: I would appreciate an advice from people who use cryo-immuno diamond knife from Diatome for the Tokuyasu technique. It is a newer model than their cryo diamond knife. Both can be seen at the following web site. http://www.emsdiasum.com/diatome/knife/immunocytochemistry.htm
Is the newer model indeed better than the older one?
Is there somebody in San Diego area who uses the cryo immuno knife?
Thank you very much,
Halina

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From: rra-at-stowers-institute.org
Date: Tue, 3 Jan 2006 20:11:12 -0600
Subject: [Microscopy] viaWWW: glue to make ribbon's stick together

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Email: rra-at-stowers-institute.org
Name: Rhonda Allen

Organization: Stowers Institute

Title-Subject: [Filtered] glue to make ribbon's stick together

Question: Hello,
I was wondering if any of you could tell me what kind of glue is used to make spurr's ribbon's stick together. I need to do serial sections and collect them all. Since I am new to this I am open for any and all suggestions.
Thanks in adance.
Rhonda Allen
rra-at-stowers-institute.org

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

==============================Original Headers==============================
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From: tiekotte-at-up.edu
Date: Tue, 3 Jan 2006 20:31:18 -0600
Subject: [Microscopy] Zeiss EM 900

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Can someone please send me the information list from a fellow lister
regarding the availability of a Zeiss EM 900?

Thank you and have great 2006.

Kenneth L. Tiekotter, Adjunct Professor
The University of Portland
Department of Biology
5000 N Willamette Blvd.
Portland, OR 97203 USA

Tel.: 503.943.8861
Email: tiekotte-at-up.edu


==============================Original Headers==============================
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5, 18 -- Date: Tue, 03 Jan 2006 18:30:35 -0800
5, 18 -- Subject: Zeiss EM 900
5, 18 -- From: Ken Tiekotter {tiekotte-at-up.edu}
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5, 18 -- Message-ID: {BFE0734B.648B%tiekotte-at-up.edu}
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From: tiekotte-at-up.edu
Date: Wed, 4 Jan 2006 00:48:06 -0600
Subject: [Microscopy] Zeiss EM900

Contents Retrieved from Microscopy Listserver Archives
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Thank you all who responded to my request for info from the list regarding
the Zeiss EM900.

Sincerely,
Ken

The University of Portland
Department of Biology
5000 N Willamette Blvd.
Portland, OR 97203 USA

Tel.: 503.943.8861
Email: tiekotte-at-up.edu


==============================Original Headers==============================
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From: gwe-at-ufl.edu
Date: Wed, 4 Jan 2006 16:03:59 -0600
Subject: [Microscopy] Re: viaWWW: glue to make ribbon's stick together

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

I seem to recall that diluted rubber cement can be used

rra-at-stowers-institute.org wrote:
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} Email: rra-at-stowers-institute.org
} Name: Rhonda Allen
}
} Organization: Stowers Institute
}
} Title-Subject: [Filtered] glue to make ribbon's stick together
}
} Question: Hello,
} I was wondering if any of you could tell me what kind of glue is used to make spurr's ribbon's stick together. I need to do serial sections and collect them all. Since I am new to this I am open for any and all suggestions.
} Thanks in adance.
} Rhonda Allen
} rra-at-stowers-institute.org
}
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}
} ==============================Original Headers==============================
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--
Greg Erdos
5410 SE 185th Ave.
Micanopy, FL 32667

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From: tsoumt-at-ilan.tpg.gov.tw
Date: Thu, 5 Jan 2006 17:20:34 -0600
Subject: [Microscopy] viaWWW: Canon pro1 onto trinocular port of Olympus BX51

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Email: tsoumt-at-ilan.tpg.gov.tw
Name: Jack Tsou

Organization: I-lan Hospital, Taiwan

Title-Subject: [Filtered] Has anyone successfully mounted Canon pro1 onto trinocular port of Olympus BX51?

Question: I'm quite interested in the digital camera Canon pro1 and would like to know the detail if anybody had ever successfully mounted it onto the trinocular port of BX51. Thanks in advance!

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6, 12 -- Subject: viaWWW: Canon pro1 onto trinocular port of Olympus BX51
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From: edelmare-at-muohio.edu
Date: Fri, 6 Jan 2006 08:33:41 -0600
Subject: [Microscopy] Re: viaWWW: Canon pro1 onto trinocular port of Olympus BX51

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

Jack:

If you haven't committed to the Canon Powershot Pro1 yet, I
strongly recommend you move up to a camera with a removable
lens like the Canon EOS 350D or The Nikon D50 - and use the
Microscope as the lens.



On 5 Jan 2006, at 19:57, tsoumt-at-ilan.tpg.gov.tw wrote:

}
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} Email: tsoumt-at-ilan.tpg.gov.tw
} Name: Jack Tsou
}
} Organization: I-lan Hospital, Taiwan
}
} Title-Subject: [Filtered] Has anyone successfully mounted Canon pro1
} onto trinocular port of Olympus BX51?
}
} Question: I'm quite interested in the digital camera Canon pro1 and
} would like to know the detail if anybody had ever successfully mounted
} it onto the trinocular port of BX51. Thanks in advance!
}
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} ==============================Original
} Headers============================== 6, 12 -- From
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} 6, 12 -- Subject: viaWWW: Canon pro1 onto trinocular port of Olympus
} BX51 6, 12 -- Content-Type: text/plain; charset="us-ascii"
} ==============================End of -
} Headers==============================



Richard E. Edelmann, Ph.D.
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

"WE ARE MICROSOFT.
RESISTANCE IS FUTILE.
YOU WILL BE ASSIMILATED."

==============================Original Headers==============================
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From: wadowska-at-upei.ca
Date: Fri, 6 Jan 2006 11:00:21 -0600
Subject: [Microscopy] LM tungsten carbide knives

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

Hello everybody,
One of our EM lab users wants to buy triangular tungsten carbide
knives for serial sectioning. Is there any advantage in buying that
kind of knife? What is a life span (how many blocks/sections can be
cut before it becomes dull)? Can they be resharpen? Are they good
for EM resins (Epon, Spurrs ect.)?
Thanks
Dorota

==============================Original Headers==============================
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From: ahlst007-at-umn.edu
Date: Fri, 6 Jan 2006 11:03:20 -0600
Subject: [Microscopy] Re: viaWWW: glue to make ribbon's stick together

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

Rhonda,

For glue to get section ribbons to stick
together, I like to use Weldwood Contact Cement
(avialable at your local hardware store) diluted
1:1 in xylene. I picked up this tip somewhere a
few years ago, but don't remember where. Maybe
it was from Microscopy!

I assume you have trimed the block to be a 4
sided pyramid with a flat top being the
blockface from which sections will be cut. To
apply glue, use a small end of a toothpick, dip
in diluted glue, and dab onto one of the 4 sides
of the block face, either top or bottom relative
to orientation of block during sectioning. I
usually tilt the block face so that the side I
choose is nearly horizontal so the glue doesn't
run down the block side, or over the actual face
of the block. Be careful so that glue does not
get onto the blockface itself. Also, work
quickly as the glue will dry fast. Then reorient
the block for sectioning.

There is also a product out there called Tacky
Wax, available from EM supply companies, which
you dab onto the side of the block face. You may
or may not need to apply gentle heat near it to
get it to spread evenly over the side of the
blockface. Perhaps others may wish to comment on
the use of Tacky Wax.

Hope this helps you!

Gib
----
Gib Ahlstrand
Imaging Center
University of Minbnesota
St. Paul, MN 55108
ahlst007-at-umn.eud
http://www.cbs.umn.edu/ic

rra-at-stowers-institute.org wrote:
} Email: rra-at-stowers-institute.org
} Name: Rhonda Allen
}
} Organization: Stowers Institute
}
} Title-Subject: [Filtered] glue to make ribbon's stick together
}
} Question: Hello,
} I was wondering if any of you could tell me what kind of glue is used to make spurr's ribbon's stick together. I need to do serial sections and collect them all. Since I am new to this I am open for any and all suggestions.
} Thanks in adance.
} Rhonda Allen
} rra-at-stowers-institute.org

==============================Original Headers==============================
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From: pmrice-at-almaden.ibm.com
Date: Fri, 6 Jan 2006 11:24:00 -0600
Subject: [Microscopy] TEM - Job Opening for TEM Specimen Preparation Lab Technician

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

Job Opening #B005608; Lab Technician/Engineer at IBM

There is an opening at the Lab Technician/Engineer level within the
Materials Characterization and Analysis Group
in the Research Division of IBM at the Almaden Research Center in San
Jose, California.
The job is a long term (three years) supplemental position with
benefits.This position involves technical support in
an advanced electron microscopy laboratory which emphasizes transmission
electron microscopy. The technician
works as a team member with professional (Ph.D.) microscopists and other
scientists in a research laboratory.
Duties involve primarily sample preparation methods including the
conventional grinding, polishing, and ion milling techniques,
focused ion beam (FIB ) method, as well as Microtome, etc. In addition,
the candidate must be able to operate
and perform routine maintenance on specimen preparation equipment, interact
with customers (professional scientists),
and prepare reports. The candidate must be able to work independently
within set priorities, to keep abreast of new advances,
and to interact smoothly within a team.

Experience with sample preparation and basic TEM imaging is a plus.

IBM is an equal opportunity employer. Women and Minorities are encouraged
to apply.
Information on the Almaden Research Center can be found at
www.almaden.ibm.com

Interested parties may call or send resumes to me at the following address:

Dr. Philip M. Rice
IBM Research Division
Almaden Research Center
650 Harry Road, K19B/D1
tel: (408) 927-1442
fax: (408) 927-2100
email: pmrice-at-almaden.ibm.com





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From: gsosinsky-at-ucsd.edu
Date: Sat, 7 Jan 2006 12:47:34 -0600
Subject: [Microscopy] viaWWW: Fourth International Congress on Electron Tomography

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

This Question/Comment was submitted to the Microscopy Listserver
using the WWW based Form at http://www.microscopy.org/MicroscopyListserver/MLFormMail.html
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Email: gsosinsky-at-ucsd.edu
Name: Gina Sosinsky

Organization: University of California, San Diego

Title-Subject: [Filtered] 4th International Congress for Electron Tomography 2006

Question: **************** First Announcement ***************
The Fourth International Congress on Electron Tomography, (4ICET), will be held at Paradise Point, San Diego November 5-8, 2006.

This congress brings together biologists, biophysicists, computer scientists, mathematicians, materials scientists and electron optical instrumentation specialists for an interdisciplinary exchange of ideas focusing on advancing methods of electron tomography (ET) in biology. ET has moved from a specialized experimental technique practiced by a few laboratories to one delivering critical information to a broad community of cell biologists, structural biologists, and neuroscientists. For students, this conference presents a unique opportunity to learn about cutting-edge advances in ET applications and methodologies.

Sessions will include:

ļ Imaging of dynamic structures and correlative microscopy Co-Chairs: O. Medalia (Ben-Gurion Univ.); Jack Johnson (The Scripps Research Institute)

ļ Advances in instrumentation
Co-chairs: M. Ellisman (UCSD); Bram Koster (Univ. ofUltrect, Netherlands)

ļ 3D reconstruction algorithms
Co-chairs: N. Volkmann (Burnham Institute); TBA

ļ Visualization & quantitative analysis
Co-chairs: R. Whitaker (Univ. of Utah); D. Mastronarde (Univ. of Colorado)

ļ Moving tomography to the mainstream: data sharing, data integration, & model building
Co-chairs: M. Martone (UCSD); J-M. Carazo (Univ. of Madrid)

ļ Emerging technologies for the multiscale: cell to tissue and molecule to cell
Co-chairs: D. Hanein (Burnham Institute); C. Larabell (UCSF)

Sessions will include both invited speakers and talks selected from submitted abstracts.

For further information please check out our web site:
http://www.4icet.org

or contact Mark H. Ellisman, Lead Congress Organizer (mark-at-ncmir.ucsd.edu) or Grace Osborne, Congress Coordinator (gosborne-at-ncmir,ucsd.edu)

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


==============================Original Headers==============================
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From: gsosinsky-at-ucsd.edu
Date: Sat, 7 Jan 2006 12:59:49 -0600
Subject: [Microscopy] viaWWW: Fourth International Congress on Electron Tomography

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

This Question/Comment was submitted to the Microscopy Listserver
using the WWW based Form at http://www.microscopy.org/MicroscopyListserver/MLFormMail.html
---------------------------------------------------------------------------
Remember this posting is most likely not from a Subscriber, so when replying
please copy both gsosinsky-at-ucsd.edu as well as the MIcroscopy Listserver
---------------------------------------------------------------------------

Email: gsosinsky-at-ucsd.edu
Name: Gina Sosinsky

Organization: University of California, San Diego

Title-Subject: [Filtered] 4th International Congress for Electron Tomography 2006

Question: **************** First Announcement ***************
The Fourth International Congress on Electron Tomography, (4ICET), will be held at Paradise Point, San Diego November 5-8, 2006.

This congress brings together biologists, biophysicists, computer scientists, mathematicians, materials scientists and electron optical instrumentation specialists for an interdisciplinary exchange of ideas focusing on advancing methods of electron tomography (ET) in biology. ET has moved from a specialized experimental technique practiced by a few laboratories to one delivering critical information to a broad community of cell biologists, structural biologists, and neuroscientists. For students, this conference presents a unique opportunity to learn about cutting-edge advances in ET applications and methodologies.

Sessions will include:

ļ Imaging of dynamic structures and correlative microscopy Co-Chairs: O. Medalia (Ben-Gurion Univ.); Jack Johnson (The Scripps Research Institute)

ļ Advances in instrumentation
Co-chairs: M. Ellisman (UCSD); Bram Koster (Univ. ofUltrect, Netherlands)

ļ 3D reconstruction algorithms
Co-chairs: N. Volkmann (Burnham Institute); TBA

ļ Visualization & quantitative analysis
Co-chairs: R. Whitaker (Univ. of Utah); D. Mastronarde (Univ. of Colorado)

ļ Moving tomography to the mainstream: data sharing, data integration, & model building
Co-chairs: M. Martone (UCSD); J-M. Carazo (Univ. of Madrid)

ļ Emerging technologies for the multiscale: cell to tissue and molecule to cell
Co-chairs: D. Hanein (Burnham Institute); C. Larabell (UCSF)

Sessions will include both invited speakers and talks selected from submitted abstracts.

For further information please check out our web site:
http://www.4icet.org

or contact Mark H. Ellisman, Lead Congress Organizer (mark-at-ncmir.ucsd.edu) or Grace Osborne, Congress Coordinator (gosborne-at-ncmir,ucsd.edu)

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


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From: jbpawley-at-wisc.edu
Date: Fri, 6 Jan 2006 12:55:00 -0600
Subject: [Microscopy] First announcement: Eleventh International UBC Live-cell Course.

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

Hello all,

Year 10 was last year: I hope you can join us as the UBC Course
enters its second decade.

Speaking for myself, and I am sure for our faculty, it is a source of
great satisfaction to see how many of the contributors to this list
are UBC Course Alumnae.

Thank you all.

Jim P.

Eleventh Annual INTERNATIONAL 12-Day Short Course on 3D Microscopy of
Living Cells, June 10 - 22, 2006 (Pre-course: June 10)

Tenth, Post-course Workshop on 3D Image Processing, June 24 -26, 2006

Organized by Prof. James Pawley, (University of Wisconsin-Madison)

in association with the Departments of Pharmacology and Physiology
and the Brain Research Centre,
University of British Columbia, Vancouver, BC, Canada

DATES

Applications must be received by Tuesday, March 15, 2006
Deposit due Friday, April 15, 2006
Registration 5:00 - 7:00 PM Saturday, June 10, 2006
First Lecture 7:30 PM Saturday, June 10, 2006
Live-cell Course ends, noon Thursday, June 22, 2006
3D Image Processing Course, Saturday, June 24 - Monday, 26, 2006


APPLICATIONS DUE BY MARCH 15, 2006

APPLICATIONS
Applicants must complete a questionnaire on the web to assess
knowledge level, field of interest and proposed personal, live-cell,
project. But don't let this put you off: if you plan to use 3D
microscopy on living cells, we can usually find a way to make it work.

Enrollment will be limited to about 32 participants (exact number
depends on number of 3D Systems available). Selection will be made
on the basis of background and perceived need. Those without
previous LM experience will be provided with access to basic texts to
read before the course begins. Application forms may be down-loaded
from the WWW site at

http://www.3dcourse.ubc.ca/application.htm , or obtained from:

Prof. James B. Pawley, Ph.
608-263-3147
Room 223, Zoology Research Building, FAX 608-265-5315
250 N. Mills Street, Madison, WI, 53706 JBPAWLEY-at-WISC.EDU

Additional information is available from:
http://www.3dcourse.ubc.ca/brochure.htm and links.

We expect to have at least 11, 3D microscope workstations for student
use and there will be an international faculty of 20.

Application deadlines:

Application forms should be received for screening by March 15, 2006.
Successful applicants will be notified by April 1, and a deposit of
50% must be received by April 15, 2006. In general, refunds of the
deposit will only be possible if someone on the waiting list can take
the place but this has not been a problem in previous years. The
remaining balance is due before Registration.


Pre-Course Tuition (1/2 day Basic Optical principles)
$125 (US)
3D Live-cell Course Tuition (includes lunches, snacks, 3 dinners, incl. the
NEW Third Edition of the Handbook of Biological Confocal
Microscopy): $2,750 (US)
Workshop Tuition (includes lunches and snacks):
$1,100 (US)

Room/board about $40/day (US) depending on room type.

I hope that this includes all of the information that you need, but
if not, please get back to me.

Jim Pawley

--
****************************************
Prof. James B. Pawley, Ph. 608-263-3147
Room 223, Zoology Research Building, FAX 608-262-9083
250 N. Mills St., Madison, WI, 53706 JBPAWLEY-at-WISC.EDU
"A scientist is not one who can answer questions but one who can
question answers." Theodore Schick Jr., Skeptical Enquirer, 21-2:39

==============================Original Headers==============================
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From: YANGA-at-AGR.GC.CA
Date: Mon, 9 Jan 2006 09:25:19 -0600
Subject: [Microscopy] Re: viaWWW: glue to make ribbon's stick together

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

In our lab, there was a person using a thin coat of dental wax successfully on bottom side of a block. Melt dental in a beaker and apply a tiny bit to a shaped block with a warm spatula. You may have to trim out excess wax.

I have Tackiwax also. It came in a tiny bottle. It seems to me that it is repackaged dental wax without color. On the other hand, I may be wrong this material.

Ann Fook Yang
EM Unit/ Unite EM
AAFC/AAC
960 Carling Ave,
Ottawa,Ontario
Canada K1A 0C6
yanga-at-agr.gc.ca
Telephone/Téléphone: 613-759-1638
Facsimile/Télécopieur: 613-759-1701

Agriculture and Agri-Food Canada - Agriculture et Agroalimentaire Canada

-----Original Message-----
X-from: ahlst007-at-umn.edu [mailto:ahlst007-at-umn.edu]
Sent: Friday, January 06, 2006 5:35 PM
To: Yang, Ann-Fook

Rhonda,

For glue to get section ribbons to stick
together, I like to use Weldwood Contact Cement
(avialable at your local hardware store) diluted
1:1 in xylene. I picked up this tip somewhere a
few years ago, but don't remember where. Maybe
it was from Microscopy!

I assume you have trimed the block to be a 4
sided pyramid with a flat top being the
blockface from which sections will be cut. To
apply glue, use a small end of a toothpick, dip
in diluted glue, and dab onto one of the 4 sides
of the block face, either top or bottom relative
to orientation of block during sectioning. I
usually tilt the block face so that the side I
choose is nearly horizontal so the glue doesn't
run down the block side, or over the actual face
of the block. Be careful so that glue does not
get onto the blockface itself. Also, work
quickly as the glue will dry fast. Then reorient
the block for sectioning.

There is also a product out there called Tacky
Wax, available from EM supply companies, which
you dab onto the side of the block face. You may
or may not need to apply gentle heat near it to
get it to spread evenly over the side of the
blockface. Perhaps others may wish to comment on
the use of Tacky Wax.

Hope this helps you!

Gib
----
Gib Ahlstrand
Imaging Center
University of Minbnesota
St. Paul, MN 55108
ahlst007-at-umn.eud
http://www.cbs.umn.edu/ic

rra-at-stowers-institute.org wrote:
} Email: rra-at-stowers-institute.org
} Name: Rhonda Allen
}
} Organization: Stowers Institute
}
} Title-Subject: [Filtered] glue to make ribbon's stick together
}
} Question: Hello,
} I was wondering if any of you could tell me what kind of glue is used to make spurr's ribbon's stick together. I need to do serial sections and collect them all. Since I am new to this I am open for any and all suggestions.
} Thanks in adance.
} Rhonda Allen
} rra-at-stowers-institute.org

==============================Original Headers==============================
7, 20 -- From ahlst007-at-umn.edu Fri Jan 6 11:02:56 2006
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7, 20 -- Date: Fri, 06 Jan 2006 11:02:39 -0600
7, 20 -- From: Gib Ahlstrand {ahlst007-at-umn.edu}
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7, 20 -- Subject: Re: [Microscopy] viaWWW: glue to make ribbon's stick together
7, 20 -- References: {200601040322.k043ML3t002256-at-ns.microscopy.com}
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From: David.Patton-at-uwe.ac.uk
Date: Mon, 9 Jan 2006 09:32:46 -0600
Subject: [Microscopy] Re: viaWWW: glue to make ribbon's stick together

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

Does anyone know if the various glues suggested vaporise in the TEM? If so does this cause any problems?

Dave

-----Original Message-----
X-from: YANGA-at-AGR.GC.CA [mailto:YANGA-at-AGR.GC.CA]
Sent: 09 January 2006 15:28
To: David Patton

In our lab, there was a person using a thin coat of dental wax successfully on bottom side of a block. Melt dental in a beaker and apply a tiny bit to a shaped block with a warm spatula. You may have to trim out excess wax.

I have Tackiwax also. It came in a tiny bottle. It seems to me that it is repackaged dental wax without color. On the other hand, I may be wrong this material.

Ann Fook Yang
EM Unit/ Unite EM
AAFC/AAC
960 Carling Ave,
Ottawa,Ontario
Canada K1A 0C6
yanga-at-agr.gc.ca
Telephone/TƩlƩphone: 613-759-1638
Facsimile/TƩlƩcopieur: 613-759-1701

Agriculture and Agri-Food Canada - Agriculture et Agroalimentaire Canada

-----Original Message-----
X-from: ahlst007-at-umn.edu [mailto:ahlst007-at-umn.edu]
Sent: Friday, January 06, 2006 5:35 PM
To: Yang, Ann-Fook

Rhonda,

For glue to get section ribbons to stick
together, I like to use Weldwood Contact Cement
(avialable at your local hardware store) diluted
1:1 in xylene. I picked up this tip somewhere a
few years ago, but don't remember where. Maybe
it was from Microscopy!

I assume you have trimed the block to be a 4
sided pyramid with a flat top being the
blockface from which sections will be cut. To
apply glue, use a small end of a toothpick, dip
in diluted glue, and dab onto one of the 4 sides
of the block face, either top or bottom relative
to orientation of block during sectioning. I
usually tilt the block face so that the side I
choose is nearly horizontal so the glue doesn't
run down the block side, or over the actual face
of the block. Be careful so that glue does not
get onto the blockface itself. Also, work
quickly as the glue will dry fast. Then reorient
the block for sectioning.

There is also a product out there called Tacky
Wax, available from EM supply companies, which
you dab onto the side of the block face. You may
or may not need to apply gentle heat near it to
get it to spread evenly over the side of the
blockface. Perhaps others may wish to comment on
the use of Tacky Wax.

Hope this helps you!

Gib
----
Gib Ahlstrand
Imaging Center
University of Minbnesota
St. Paul, MN 55108
ahlst007-at-umn.eud
http://www.cbs.umn.edu/ic

rra-at-stowers-institute.org wrote:
} Email: rra-at-stowers-institute.org
} Name: Rhonda Allen
}
} Organization: Stowers Institute
}
} Title-Subject: [Filtered] glue to make ribbon's stick together
}
} Question: Hello,
} I was wondering if any of you could tell me what kind of glue is used to make spurr's ribbon's stick together. I need to do serial sections and collect them all. Since I am new to this I am open for any and all suggestions.
} Thanks in adance.
} Rhonda Allen
} rra-at-stowers-institute.org

==============================Original Headers==============================
7, 20 -- From ahlst007-at-umn.edu Fri Jan 6 11:02:56 2006
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From: M_Jarnik-at-fccc.edu
Date: Mon, 9 Jan 2006 10:03:33 -0600
Subject: [Microscopy] His Tag antibody for EM

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

I posted this question a couple of weeks ago, but probably the timing was wrong (just before the holidays), no answers at all. So I am posting again:

I would need a tip on a good (commercially available) antibody against His tag for EM - Tokuyasu technique. It should tolerate at least light (0.1-0.2%) glutaraldehyde fixation. A rabbit polyclonal would be preferable, but a working monoclonal would do as well.

Another unrelated question - I would need to label macrophages in sections (again, Tokuyasu). Any idea for a good macrophage marker?

Thanks,

Michal



==============================Original Headers==============================
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From: mjo10-at-psu.edu
Date: Mon, 9 Jan 2006 18:23:39 -0600
Subject: [Microscopy] viaWWW: Vanadium oxide thin films

Contents Retrieved from Microscopy Listserver Archives
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Name: Matthew Olszta

Organization: Penn State University

Title-Subject: [Filtered] Vanadium oxide thin films

Question: I wanted to know if anyone has had any experience with TEM sample preparation and analysis of vanadium oxide thin films. The reason I ask is that certain vanadium oxide crystal structures (VO2 and V2O5) are soluble or slightly soluble in water, but using water in polishing I have not yet seen any appreciable dissolution in my cross-sections.

If others have worked with these materials, or other water sensitive materials, do you have any suggestions or comments on sample preparation in order to avoid artifacts?

Regards and advanced thanks,
Matt



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From: mjo10-at-psu.edu
Date: Mon, 9 Jan 2006 18:32:46 -0600
Subject: [Microscopy] viaWWW: Vanadium oxide thin films

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Email: mjo10-at-psu.edu
Name: Matthew Olszta

Organization: Penn State University

Title-Subject: [Filtered] Vanadium oxide thin films

Question: I wanted to know if anyone has had any experience with TEM sample preparation and analysis of vanadium oxide thin films. The reason I ask is that certain vanadium oxide crystal structures (VO2 and V2O5) are soluble or slightly soluble in water, but using water in polishing I have not yet seen any appreciable dissolution in my cross-sections.

If others have worked with these materials, or other water sensitive materials, do you have any suggestions or comments on sample preparation in order to avoid artifacts?

Regards and advanced thanks,
Matt



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From: hall-at-aecom.yu.edu
Date: Tue, 10 Jan 2006 10:06:01 -0600
Subject: [Microscopy] glue to make ribbon's stick together

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

I am not sure about vaporization issues, but I have used another
glue. Using a sharpened toothpick dipped in acetone, I have
dissolved glue from 3M adhesive tape ("Magic Tape") to apply thin
amounts of glue to the top surface of the block when trying to obtain
serial sections. I have not noted any contamination problems, but
clearly you want to refrain from getting any of this solvent on the
cutting face.

However, when collecting serial sections, glue should only be your
last resort. When properly trimmed with a fresh razor blade (I like
the chromium steel blades from "Wilkinson Sword Blades") so that both
top edge and bottom edge are parallel to each other and to the
diamond knife edge, most Epon-like resins allow serial sections to
stick together quite well. I don't know how well this applies to
Spurr's in particular, but I expect the same results. The cleaner
those edges are, and the more parallel they are to each other and to
the diamond, the better the result. Acrylic resins like LR White or
LR Gold tend to be much more brittle, and those would be more
difficult to handle in series.

Dave Hall

For a more exhaustive discussion of serial sectioning, see Hall
(1995) Methods in Cell Biology, C. elegans: Modern Biological
Analysis of an Organism. Epstein and Shakes (eds) Academic Press.
pp. 395-436.
--
David H. Hall, Ph.D.
Center for C. elegans Anatomy
Department of Neuroscience
Albert Einstein College of Medicine
1410 Pelham Parkway
Bronx, NY 10461

www.wormatlas.org
www.aecom.yu.edu/wormem

phone 718 430-2195
fax 718 430-2514

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From: mkamrath-at-nalco.com
Date: Tue, 10 Jan 2006 18:27:47 -0600
Subject: [Microscopy] viaWWW: Colloidal Silica Particle Size Distribution by TEM

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

Matt,
I don't have experience to work with vanadium oxide, but some
superconductive materials which were not working well with water. If you use
water only for your grinding procedure for your case, now you can try
dry-grinding, at least for the final stage. or you can try leaving thicker
for the final ion mill (like 45 micron) if it is not hard to be ion-milled
under Ar+ ion beam, but this may cause some beam damage or contamination if
there is not cold stage and the milling time is too long. again to try
higher rate for the early milling stage and to reduce it at the final may
help with this. or you could try methanol or ethanol for your grinding if
the "wet" procedure is necessary. also you can try FIB. it is dry, but may
consider its beam damage if your material is sensitive to this.
Hope it can help




Sincerely
Long Li
_________________________________________________________
Materials Science and Engineering Department
University of Pittsburgh
3700 O'Hara St., 848 Benedum Hall
Pittsburgh, PA 15261

Tel: (412) 624-9753
FAX: (412) 624-8069
e-mail: Lil2-at-pitt.edu



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Email: mkamrath-at-nalco.com
Name: Mike Kamrath

Organization: Nalco Co., Naperville IL

Title-Subject: [Filtered] Colloidal Silica Particle Size Distribution by TEM

Question: I was looking for help with a problem we have related to particle size distributions (PSD) with colloidal silicas as measured by TEM. We normally dilute a colloidal silica solution in an aqueous solution containing a few drops of nonionic surfactant to help disperse the particles (5-100 nm). After placing a drop on a copper grid (Formvar) we dry it at 100 oC for 10 minutes and place in the TEM for PSD. We are trying to automate the process with counting software but the software has problems identifying individual particles when they are touching one another. I have two questions related to this: 1) Does anyone know of software capable of easily separating touching particles and counting individually? We are currently using Image-Pro Plus and I am aware of free software from NIH (Scion) which seems to have similar problems. 2) Does anybody have suggestions for a better solvent system or surfactant that might separate the particles better? Or is the drying process forcing the particles together and would a vacuum drying or other method work better?

Thanks in advance for your help.

Mike Kamrath
Nalco Co.
Naperville, IL



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From: r.sims-at-auckland.ac.nz
Date: Tue, 10 Jan 2006 19:00:57 -0600
Subject: [Microscopy] Oil on detector window

Contents Retrieved from Microscopy Listserver Archives
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Here's a problem for the New Year:

A couple of months ago, I repumped in situ my chronically leaky PGT Be-window EDS
detector, and ran 50 deg C air into the dewar while re-evacuating using the chamber
vacuum via a hose from the sample introduction port of the JEOL 840 on which the
detector is mounted.

The re-evacuation worked well, in that the LN2 consumption is now back to normal, but
after recooling and switching on the bias again, there was no appreciable low-energy
response -- no visible Cu L peak at all!

I realised that grease from the O-ring sliding seal through which the tube of the detector
enters the chamber had melted and run down onto the Be window. Cleaning of the
window with Freon restored the Cu L intensity, and all was well, until it became obvious
that the window was re-oiling again at a much faster rate than ever before.

Because we do quantitative mineral analysis, I keep a good handle on the window
condition, evidenced by both Cu L and Na K responses. I have been using this detector
for about five years and not found it neccesary to clean the window before, but now the
Na response halves in a few weeks. Recleaning with Freon restores the response
again.

In an effort to fix this, I have:-

- changed the rotary pump oil
- installed an alumina-pellet foreline trap
- changed the DP oil (Santovac, but the old charge was still light-colored)

but still the darned window is oiling up.

I'm running out of ideas. It seems that either the chamber atmosphere has become
markedly oilier than before, or the detector window is now running colder than before.

My money is on the latter, as it seems too much of a coincidence for some change in
the back-streaming performance of the 840 to have occurred at the same time as the
repumping of the detector, but I can't see what might have happened.

Anyone got any suggestions?

Happy New Year

rtch


--
Ritchie Sims Ph D Phone : 64 9 3737599 ext 87713
Microanalyst Fax : 64 9 3737435
Department of Geology email : r.sims-at-auckland.ac.nz
The University of Auckland
Private Bag 92019
Auckland
New Zealand


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From: jrminter-at-rochester.rr.com
Date: Tue, 10 Jan 2006 22:25:01 -0600
Subject: [Microscopy] viaWWW: Colloidal Silica Particle Size Distribution by

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

First, may I ask what you have done to ensure that your support films are
hydrophilic? Carbon coated polyvinylformal supports become increasingly
hydrophobic with age. We get good results by treating with spectro-grade
acetone for 30 min prior to use. We have also used a short plasma-ash in air
to improve wettability. Such treatments improves dispersion and reduces
drying artifacts.

We use the analySIS Five software from Soft Imaging System for image
analysis. This program has a useful separator based upon a watershed
algorithm. I have also written a custom watershed module for some
applications. We also use a guard frame to eliminate bias from the higher
probability that larger particles touch the image boundaries compared to
smaller particles. We prefer this approach to the Miles-Lantejoul algorithm.
Using an extension to the analySIS Automater we can queue up images from
several samples from any of our microscopes and let them run unattended.

We have compared results obtained by simply deleting agglomerates with the
results obtained by separation. Even if we use a separator, we typically
measure the maximum intrusion distance [this is a custom module we wrote]
for each particle and flag those which are not convex as agglomerates. We
always keep track of the area fraction of agglomerates and reject analyses
where this is too high. Usually we can find preparation conditions to keep
the area fraction of agglomerates below 10%.

One of the strengths of the analySIS program is that the macro language is
almost a full subset of C and if one needs a computationally-intensive
function (like the maximum intrusion distance) it is easy to import a C
function from a custom DLL compiled with a good optimizing compiler, like
Visual C++.

Disclaimer: I have no financial interest in Soft Imaging System, writing
only as a satisfied user.

John R. Minter
Eastman Kodak Co.
Research Laboratories
Foundation Science and Technology Center
john.r.minter-at-kodak.com (work)
jrminter-at-rochester.rr.com (home)



-----Original Message-----
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Email: mkamrath-at-nalco.com
Name: Mike Kamrath

Organization: Nalco Co., Naperville IL

Title-Subject: [Filtered] Colloidal Silica Particle Size Distribution by TEM

Question: I was looking for help with a problem we have related to particle
size distributions (PSD) with colloidal silicas as measured by TEM. We
normally dilute a colloidal silica solution in an aqueous solution
containing a few drops of nonionic surfactant to help disperse the particles
(5-100 nm). After placing a drop on a copper grid (Formvar) we dry it at
100 oC for 10 minutes and place in the TEM for PSD. We are trying to
automate the process with counting software but the software has problems
identifying individual particles when they are touching one another. I have
two questions related to this: 1) Does anyone know of software capable of
easily separating touching particles and counting individually? We are
currently using Image-Pro Plus and I am aware of free software from NIH
(Scion) which seems to have similar problems. 2) Does anybody have
suggestions for a better solvent system or surfactant that might separate
the particles better? Or is the drying p!
rocess forcing the particles together and would a vacuum drying or other
method work better?

Thanks in advance for your help.

Mike Kamrath
Nalco Co.
Naperville, IL



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


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From: dsherman-at-purdue.edu
Date: Wed, 11 Jan 2006 08:11:13 -0600
Subject: [Microscopy] Sonicator

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

I am looking for a simple probe-type sonicator that will be used to make
holey films.

We have used an old one in the past that is no longer available. It is used
to aerate a formvar/glycerol solution sufficiently to make very small
bubbles. When a slide is dipped into the solution, the resulting film has
holes of a size dependent on the length of time used for the sonication.

Any recommendations welcomed but hopefully I can find one on the less
expensive side.

Debby

Debby Sherman, Manager Phone: 765-494-6666
Life Science Microscopy Facility FAX: 765-494-5896
Purdue University E-mail: dsherman-at-purdue.edu
S-052 Whistler Building
170 S. University Street
West Lafayette, IN 47907
http://www3.agriculture.purdue.edu/microscopy


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From: mike.reedy-at-cellbio.duke.edu
Date: Wed, 11 Jan 2006 11:17:18 -0600
Subject: [Microscopy] viaWWW: Potassium Permanganate

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

sue-
For what it's worth, and to get in synch with anyone out there
trying tests such as i described, last week I repeated my 40 year old
MnO4 tests in water on all the liquid resin components, adding 1-2
small drops to 2.5 ml dilute transparent purple MnO4 sol'n (2.5 ml
water, 2 drops 2% KMnO4). I found that NMA instantly turns the sol'n
to clear brown (guess I'd misremembered that it made a precipitate
before), and DMP-30 does so almost as fast. All the other
components, including Araldite 506, do the same thing over periods
of 5-200 minutes, Araldite 506 clearly slower than Epon 812. Some
make a precipitate, some don't. By next day, nothing retains any
purple, all test sol'ns are brown. So the lesson is, MnO4 can act
eventually destructively on any sections or their components, given
time enough.
-mike-

} hi mike...thanks for all your help!!!
} sue


--
-mike reedy-

************************
Michael K. Reedy, M.D.
Duke Univ. Med. Center
Dept. Cell Biology, Box 3011 (for U.S. Mail)
458 Alex Sands Bldg, Research Dr (courier)
Durham, NC 27710

Office 919-668-2534
Lab 919-684-5674
Fax 919-681-9929
mike.reedy-at-cellbio.duke.edu
http://note.cellbio.duke.edu/Faculty/Research/Reedy.html

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From: Robin.Moresi-at-gd-ais.com
Date: Wed, 11 Jan 2006 11:21:28 -0600
Subject: [Microscopy] Field Emission SEM question

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


Hi everyone!

I am looking into acquiring a new field emission SEM which will be used
mainly for failure analysis of metals, but also plastics, paper, an
occasional drop of grease, and anything else that wanders through the
lab (but rarely biological samples). The field emission SEMs that I am
considering are the JEOL JSM-7000F, the ZEISS Supra 40 (VP), the Hitachi
S-4300SE/N, and the FEI Quanta 600 FEG. For microanalysis systems, I am
considering the newest versions from Thermoelectron, Oxford and EDAX. I
welcome any comments that you can offer as to the function (or
malfunction) of the above instruments and anaylsis systems, as well as
possible electro-magnetic interference issues, user interface issues,
interface problems between the SEM and microanalysis systems, and the
reliability/response time of the service representatives from the above
companies.

Thank you very much for your time!

Sincerely,

Robin Moresi
General Dynamics - Advanced Information Systems 100 Plastics Ave.
Pittsfield, MA 01201

Tel. (413) 662-2889


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From: elke.buschbeck-at-uc.edu
Date: Wed, 11 Jan 2006 13:22:43 -0600
Subject: [Microscopy] Need manual for Sorvall MT2-B ultra microtome

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

Hi Everyone,

I am looking for a manual for a SORVALL MT2-B ultra microtome. I will be
happy to pay for copying and shipping costs, but would greatly appreciate
if someone would be willing to copy their manual for me.

Thanks,
Elke

Elke Buschbeck
Biological Sciences
University of Cincinnati
Cincinnati, OH
elke.buschbeck-at-uc.edu



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From: msteglic-at-mdanderson.org
Date: Wed, 11 Jan 2006 15:43:11 -0600
Subject: [Microscopy] MT2-B Manual

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

Hi Ritch

What has happened to you has happened to many before you!

Once you reach the point of contaminating your window with RP fluid it will
have contaminated the complete vacuum system. Two solutions. EITHER take
everything apart and clean it, yes ALL the vacuum lines and column liners.
OR use a hair drier to bake the pumping lines to drive off the contamination
as best as possible, as well as cleaning the parts you are easily able to
reach.

Many times in my service technician career I have been forced to take this
route. It puts you out of action for a day or so whilst the system recovers
but in the main it seems to be a cure. BUT if you do not tackle the fore
line trap idea it will happen all over again - our chemical test said it was
RP fluid by the way.

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 7802 966067 Web www.emcourses.com

----- Original Message -----
X-from: {r.sims-at-auckland.ac.nz}
To: {protrain-at-emcourses.com}
Sent: Wednesday, January 11, 2006 1:02 AM

Will the person that wanted the MT2-B manual please contact me. I found
the one I was looking for after I deleted your post.

Mannie Steglich
msteglic-at-mdanderson.org

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From: r.sims-at-auckland.ac.nz
Date: Wed, 11 Jan 2006 17:09:47 -0600
Subject: [Microscopy] More about oil on detector window

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


Many thanks for the replies and suggestions so far, however, please note that this is a
new condition.

For the past five years the oiling has not been a problem, and it has started to occur
only since the repumping operation described below in tedious detail.

There have been no changes to the system apart from the three steps taken, one at a
time, in an attempt to fix the problem (changing the RP oil; installing the foreline trap;
and changing the DP oil), and they have made no difference.

I think it would be too much of a coincidence for the cause of the oiling to be unrelated
to the repumping operation, so I think that complex cures such as installing LN2 traps,
plasma cleaning, complete stripping and cleaning, etc, do not address the question,
which is "what has changed to accelerate the detector window oiling so greatly, and
how can I get back to the previous state?"

I had thought that perhaps a drop of melted grease might have dripped into the DP, and
be decomposing in the Santovac, which propelled me to change the Santovac, but I now
realise that this is very unlikely to be the case, as the DP is not directly beneath the
sample chamber, and even if it were, such a drop of molten grease would not make it
through the water-cooled baffle above the DP. Plus, of course, the fact that very thoroughly
cleaning both of the DPs and both of their water-cooled baffles has made no difference
at all to the problem.

Incidentally, the Santovac, after five years' continuous operation, was only a pale straw
color, boy, that stuff certainly is stable, isn't it?

I feel fairly sure that there is a simple solution to my problem, waiting out there to be
noticed, so please keep the suggestions coming.

cheers

rtch


On 10 Jan 2006 at 19:02, r.sims-at-auckland.ac.nz wrote:

}
}
}
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
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}
} Here's a problem for the New Year:
}
} A couple of months ago, I repumped in situ my chronically leaky PGT Be-window EDS
} detector, and ran 50 deg C air into the dewar while re-evacuating using the chamber
} vacuum via a hose from the sample introduction port of the JEOL 840 on which the
} detector is mounted.
}
} The re-evacuation worked well, in that the LN2 consumption is now back to normal, but
} after recooling and switching on the bias again, there was no appreciable low-energy
} response -- no visible Cu L peak at all!
}
} I realised that grease from the O-ring sliding seal through which the tube of the detector
} enters the chamber had melted and run down onto the Be window. Cleaning of the
} window with Freon restored the Cu L intensity, and all was well, until it became obvious
} that the window was re-oiling again at a much faster rate than ever before.
}
} Because we do quantitative mineral analysis, I keep a good handle on the window
} condition, evidenced by both Cu L and Na K responses. I have been using this detector
} for about five years and not found it neccesary to clean the window before, but now the
} Na response halves in a few weeks. Recleaning with Freon restores the response
} again.
}
} In an effort to fix this, I have:-
}
} - changed the rotary pump oil
} - installed an alumina-pellet foreline trap
} - changed the DP oil (Santovac, but the old charge was still light-colored)
}
} but still the darned window is oiling up.
}
} I'm running out of ideas. It seems that either the chamber atmosphere has become
} markedly oilier than before, or the detector window is now running colder than before.
}
} My money is on the latter, as it seems too much of a coincidence for some change in
} the back-streaming performance of the 840 to have occurred at the same time as the
} repumping of the detector, but I can't see what might have happened.
}
} Anyone got any suggestions?
}
} Happy New Year
}
} rtch
}
}
} --
} Ritchie Sims Ph D Phone : 64 9 3737599 ext 87713
} Microanalyst Fax : 64 9 3737435
} Department of Geology email : r.sims-at-auckland.ac.nz
} The University of Auckland
} Private Bag 92019
} Auckland
} New Zealand
}
}
} ==============================Original Headers==============================
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From: kenconverse-at-qualityimages.biz
Date: Thu, 12 Jan 2006 09:30:50 -0600
Subject: [Microscopy] More about oil on detector window

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

Dear Ritch,
Does the snout of the EDS detector feel cool to the touch, when you vent the
chamber? (Use the back of your hand to test it.) I had one detector that
oiled very badly and it felt cool to the touch, so it acted as an oil trap
for the chamber. I sent that detector to be completely rebuilt as a light
element, high resolution detector and that problem has vanished, so it was a
condition of the detector, not the SEM, since the SEM was not touched. It
may be that the insulation between the liquid nitrogen or the cooled
components and the snout of the detector has changed with your warm-up and
re-pump. I am wondering if the copper braid that conducts the cool from the
dewar to the nose of the detector has moved or is touching something.
None of this helps you in any way. You can clean the Be-window detector with
a trickle of pure ethanol or iso-propanol in the SEM, if you can get at it.
----- Original Message -----
X-from: {r.sims-at-auckland.ac.nz}
To: {mager-at-interchange.ubc.ca}
Sent: Wednesday, January 11, 2006 3:13 PM

Ritch,
This is a long shot and you probably would have noticed the symptoms, but
here goes:

I had one 840 that kept being "burped" first thing in the morning, but not
after that. The key to finding the problem was that the LEDs on the vacuum
control all cycled correctly, but it took me a while (since it only happened
once a day) to notice that the actual valve operation for V2 (at the base of
the second DP) was delayed just long enough to burp the DPs. Turns out the
solenoid valve that operated V2 had Apiezon in it (a lot) and after sitting
overnight would stick for about 2 seconds, only milliseconds longer than the
delay for V5 to rough the load lock. This problem had been intermittent for
years before I took on the service contract, but things work fine, now.

Perhaps you have a similar timing problem. Of course, the most obvious
thing was that the DPs dumped and you don't seem to have had anything that
severe happening.

Another thought: when you cleaned the DPs, did you also remove and clean
the ballast tank located between them? Perhaps it has become too
contaminated and is now backstreaming since there is only one DP between it
and the chamber.

Do you have any additional gauging to monitor the actual pressure in the
chamber? There might be some clues there, also. I'm thinking along the
lines of a slightly leaky V4.

One last thought: Is the water flowing in the correct direction? An
accidental reversal puts warm water running through the water baffles and
virtually always results in oil on the EDS detector.

Best of luck,
Ken Converse

QUALITY IMAGES
Servicing Scanning Electron Microscopes
Since 1981
474 So. Bridgton Rd.
Bridgton, ME 04009
207-647-4348
Fax 207-647-2688
kenconverse-at-qualityimages.biz
qualityimages.biz


-----Original Message-----
X-from: r.sims-at-auckland.ac.nz [mailto:r.sims-at-auckland.ac.nz]
Sent: Wednesday, January 11, 2006 6:12 PM
To: kenconverse-at-qualityimages.biz


Many thanks for the replies and suggestions so far, however, please note
that this is a
new condition.

For the past five years the oiling has not been a problem, and it has
started to occur
only since the repumping operation described below in tedious detail.

There have been no changes to the system apart from the three steps taken,
one at a
time, in an attempt to fix the problem (changing the RP oil; installing the
foreline trap;
and changing the DP oil), and they have made no difference.

I think it would be too much of a coincidence for the cause of the oiling to
be unrelated
to the repumping operation, so I think that complex cures such as installing
LN2 traps,
plasma cleaning, complete stripping and cleaning, etc, do not address the
question,
which is "what has changed to accelerate the detector window oiling so
greatly, and
how can I get back to the previous state?"

I had thought that perhaps a drop of melted grease might have dripped into
the DP, and
be decomposing in the Santovac, which propelled me to change the Santovac,
but I now
realise that this is very unlikely to be the case, as the DP is not directly
beneath the
sample chamber, and even if it were, such a drop of molten grease would not
make it
through the water-cooled baffle above the DP. Plus, of course, the fact that
very thoroughly
cleaning both of the DPs and both of their water-cooled baffles has made no
difference
at all to the problem.

Incidentally, the Santovac, after five years' continuous operation, was only
a pale straw
color, boy, that stuff certainly is stable, isn't it?

I feel fairly sure that there is a simple solution to my problem, waiting
out there to be
noticed, so please keep the suggestions coming.

cheers

rtch


On 10 Jan 2006 at 19:02, r.sims-at-auckland.ac.nz wrote:

}
}
}
}
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}
} Here's a problem for the New Year:
}
} A couple of months ago, I repumped in situ my chronically leaky PGT
Be-window EDS
} detector, and ran 50 deg C air into the dewar while re-evacuating using
the chamber
} vacuum via a hose from the sample introduction port of the JEOL 840 on
which the
} detector is mounted.
}
} The re-evacuation worked well, in that the LN2 consumption is now back to
normal, but
} after recooling and switching on the bias again, there was no appreciable
low-energy
} response -- no visible Cu L peak at all!
}
} I realised that grease from the O-ring sliding seal through which the tube
of the detector
} enters the chamber had melted and run down onto the Be window. Cleaning of
the
} window with Freon restored the Cu L intensity, and all was well, until it
became obvious
} that the window was re-oiling again at a much faster rate than ever
before.
}
} Because we do quantitative mineral analysis, I keep a good handle on the
window
} condition, evidenced by both Cu L and Na K responses. I have been using
this detector
} for about five years and not found it neccesary to clean the window
before, but now the
} Na response halves in a few weeks. Recleaning with Freon restores the
response
} again.
}
} In an effort to fix this, I have:-
}
} - changed the rotary pump oil
} - installed an alumina-pellet foreline trap
} - changed the DP oil (Santovac, but the old charge was still
light-colored)
}
} but still the darned window is oiling up.
}
} I'm running out of ideas. It seems that either the chamber atmosphere has
become
} markedly oilier than before, or the detector window is now running colder
than before.
}
} My money is on the latter, as it seems too much of a coincidence for some
change in
} the back-streaming performance of the 840 to have occurred at the same
time as the
} repumping of the detector, but I can't see what might have happened.
}
} Anyone got any suggestions?
}
} Happy New Year
}
} rtch
}
}
} --
} Ritchie Sims Ph D Phone : 64 9 3737599 ext
87713
} Microanalyst Fax : 64 9 3737435
} Department of Geology email :
r.sims-at-auckland.ac.nz
} The University of Auckland
} Private Bag 92019
} Auckland
} New Zealand
}
}
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From: eschumacher-at-mccrone.com
Date: Thu, 12 Jan 2006 14:13:17 -0600
Subject: [Microscopy] Short Course Announcement: Scientific Imaging, Transmission Electron Microscopy

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

Fellow Listers,

The College of Microscopy in Westmont, Illinois, is pleased to announce
two new short courses for 2006:

Scientific Imaging (http://www.collegeofmicroscopy.com/courses/18.asp)

Transmission Electron Microscopy
(http://www.collegeofmicroscopy.com/courses/17.asp)


Both are hands-on, introductory level courses taught using the latest
equipment available. Please follow the links provided for further
details and registration information.


Elaine Schumacher
Senior Research Scientist
McCrone Associates, Inc.
850 Pasquinelli Drive
Westmont, IL 60559-5539 USA
630-887-7100 (tel)
630-887-7417 (fax)
E-mail: eschumacher-at-mccrone.com
Web Site: www.mccrone.com




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From: TindallR-at-missouri.edu
Date: Thu, 12 Jan 2006 16:59:23 -0600
Subject: [Microscopy] Static guns?

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

Dear listers,

Does anyone know of a reasonable alternative to the venerable Zero Stat
static taming guns? We use them often but there's just got to be a
better way. I used to smuggle squirt guns onto the school bus that were
made better (the good old days), and they didn't cost over $100 for
about $1 worth of components.

Thanks and Happy New Year to all.

Randy

Randy Tindall
EM Specialist
Electron Microscopy Core Facility---We Do Small Well!
W122 Veterinary Medicine
University of Missouri
Columbia, MO 65211
Tel: (573) 882-8304
Fax: (573) 884-2227
Email: tindallr-at-missouri.edu
Web: http://www.emc.missouri.edu







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From: pmrice-at-almaden.ibm.com
Date: Thu, 12 Jan 2006 17:19:37 -0600
Subject: [Microscopy] TEM - Job opening for a TEM Specimen Preparation Lab Technician

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

Job Opening #B005608; Lab Technician/Engineer at IBM

There is an opening at the Lab Technician/Engineer level within the
Materials Characterization and Analysis Group in the Research Division
of IBM at the Almaden Research Center in San Jose, California.
The job is a long term (three years) supplemental position with benefits.
A B.S. in material science or equivalent fields, or successful completion
of a two year college program plus experience, is required.

This position involves technical support in an advanced electron
microscopy laboratory which emphasizes transmission
electron microscopy. The technician works as a team member
with professional (Ph.D.) microscopists and other scientists in
a research laboratory. Duties involve primarily sample
preparation methods including the conventional grinding,
polishing, and ion milling techniques, focused ion beam (FIB )
method, as well as Microtome, etc. In addition, the candidate
must be able to operate and perform routine maintenance on
specimen preparation equipment, interact with customers
(professional scientists), and prepare reports. The candidate
must be able to work independently within set priorities,
to keep abreast of new advances, and to interact smoothly within a team.

Experience with sample preparation and basic TEM imaging is a plus.

IBM is an equal opportunity employer. Women and Minorities are encouraged
to apply.
Information on the Almaden Research Center can be found at
www.almaden.ibm.com

Interested parties may call or send resumes to me at the following address:

Dr. Philip M. Rice
IBM Research Division
Almaden Research Center
650 Harry Road, K19B/D1
tel: (408) 927-1442
fax: (408) 927-2100
email: pmrice-at-almaden.ibm.com


Dr. Philip M. Rice
IBM Research Division
Almaden Research Center
650 Harry Road, K19B/D1
San Jose, CA 95120-6099
tel: (408) 927-1442
fax: (408) 927-2100
e-mail: pmrice-at-almaden.ibm.com


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From: phillipst-at-missouri.edu
Date: Thu, 12 Jan 2006 18:05:32 -0600
Subject: [Microscopy] bacterial permeabilization

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

I am trying to permeabilize some Haemophilus influenzae fixed in 2%
paraformaldehyde + 0.2% glutaraldehyde so I can immunostain an
intracellular protein. My first attempts to permeabilize with Triton X-100
were surprising unsuccessful. I tried both literature searches and
googling this topic but a search of "bacteria" + "permeabilization" pulls
up thousands on non-germane citations. I am interested in comments from
anybody with expertise in permeabilization, especially in regard to the
bacterial target. 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



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From: aetmicro-at-optonline.net
Date: Thu, 12 Jan 2006 20:21:13 -0600
Subject: [Microscopy] viaWWW: Philips EM 300 beam issue

Contents Retrieved from Microscopy Listserver Archives
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This Question/Comment was submitted to the Microscopy Listserver
using the WWW based Form at http://www.microscopy.org/MicroscopyListserver/MLFormMail.html
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Email: aetmicro-at-optonline.net
Name: Andrew Thelian

Title-Subject: [Filtered] Philips EM 300 beam issue

Question: Hi All,

I have hit yet another bump in the road...

I was having a brightness issue last week that i thought might be resolved with cleaning the wehnelt assembly...

Now when i energize the filament i get a very small circle on the view screen...it doesnt appear green it looks like a dim reddish color...it also seems to respond to turning the filament knob...

the condenser and deflection knobs dont cause any change...

I have removed all apertures... to see if there was a beam any where ... but no luck...

Any suggestion would be most appreciated?

Thanks :)

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

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From: dustin.adams-at-xerox.com
Date: Thu, 12 Jan 2006 20:24:32 -0600
Subject: [Microscopy] AskAMicroscopist: SEM microscopist salaries

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Email: dustin.adams-at-xerox.com
Name: Dustin Adams

Organization: Xerox Corp.

Education: Graduate College

Location: Wilsonville, OR

Title: SEM microscopist salaries

Question: Hello, I am going to be training soon to become a SEM microscopist for my company, and was curious to see what the salary amounts are for a typical person in my position. I have tried to look for this info online, but wasn't really able to find anything. If someone could provide that information or point me in the right direction; I would greatly appreciate it.

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From: secr-at-eurmicsoc.org
Date: Fri, 13 Jan 2006 07:36:29 -0600
Subject: [Microscopy] viaWWW: EMS scholarships for IMC16 Msg 2006004

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What you see is the light from filament, not e-beam. Whenever electron gun
was disturbed, in your case for Wehnelt cleaning, gun tilt must be
re-aligned, and perhaps gun shift too. Both these alignments are mechanical
on Phil. EM300. Users manual will help if TEM is functioning properly.

Green light or not, you must be able to tell whether HT is present by
watching emission current, and emission meter behavior:
a) when HT turned on/off;
b) emission setting changed while HT is on;
c) and HV setting changed while HT is on.

If HT is present, if filament glows, and if emission current behaves
normally, do
the following:
a) select lowest magnification - I believe it is SC (scan) position;
b) move all apertures from the beam pass;
c) if no beam yet, turn lens currents off one by one and you must see the
beam. Be careful not to burn the screen (keep filament under-saturated).
Just turning off C1 and C2 will likely bring the beam back, and then work
from that point on.

Vitaly Feingold
SIA
2773 Heath Line
Duluth GA 30096
Ph. 770-232-7785
Fax 770-232-1791
www.sia-cam.com

----- Original Message -----
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Sent: Thursday, January 12, 2006 9:23 PM

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Email: secr-at-eurmicsoc.org
Name: Nick Schryvers

Organization: EMS

Title-Subject: [Filtered] EMS scholarships for IMC16 Msg 2006004

Question:
The European Microscopy Society is pleased to be able to offer 4
scholarships of 500 Euro each to young researchers in support of attending
the 16th International Microscopy Congress (http://www.imc16.jp/) in
Sapporo, Japan, from Sep. 3 till Sep. 8, 2006. Applications should be sent
to the EMS secretary (secr-at-eurmicsoc.org) and include a CV and list of
publications and copies of the submitted abstract(s) at IMC16. Proof of
acceptance of the abstract(s) should follow as soon as these are available.

Deadline of the applications is February 1, 2006 (same date as abstract
submission for IMC16)

Only EMS members are eligible and priority will be given to people under the
age of 35.

This support was made possible through a generous offer from JEOL Europe
(http://www.jeoleuro.com/).

We look forward to your applications,

Nick Schryvers
EMS secretary



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From: secr-at-eurmicsoc.org
Date: Fri, 13 Jan 2006 07:37:48 -0600
Subject: [Microscopy] viaWWW: RMS Spring School in Electron Microscopy

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Email: secr-at-eurmicsoc.org
Name: Nick Schryvers

Organization: EMS

Title-Subject: [Filtered] Royal Microscopical Society ( RMS) Spring School in Electron Microscopy

Royal Microscopical Society ( RMS) Spring School in Electron Microscopy
27-31 March 2006
University of Leeds, UK.
Course Organiser: Prof. Rik Brydson


The RMS School consists of a number of parallel courses covering most aspects of electron microscopy in both the physical and life sciences.
Students can attend either 4 or 5 days - the 5 day course includes a day of principles of EM lectures for beginners or as a refresher. During the week, students will have the opportunity to choose to attend lectures and workshops from either course, to tailor it to their specific needs.

'All those involved in the organisation of this event are to be congratulated. The complexity of running a school of electron microscopy covering both life and materials sciences has been surpassed with remarkable success with participants, speakers and organisers equally satisfied with the end results.'

Pedro Costa - Participant Spring School in EM 2005


For more information or to book for this course please visit the RMS website: {http://www.rms.org.uk/event_em_school.shtml} http://www.rms.org.uk/event_em_school.shtml


Victoria Lee
Conference Manager
Royal Microscopical Society
37/38 St Clements, Oxford OX4 1AJ

Tel: +44 (0)1865 254769
Fax: +44 (0)1865 791237
Web: {http://www.rms.org.uk} www.rms.org.uk


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From: microscopytoday-at-tampabay.rr.com
Date: Fri, 13 Jan 2006 08:32:19 -0600
Subject: [Microscopy] Re: AskAMicroscopist: SEM microscopist salaries

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Hi Randy,

I always used a sealed Polonium-210 beta source for static problems -
Nuclepore Corp used to make a Polonium-210 (sealed in beads) metal strip
that had an open ladder arrangement at the top that you simply placed the
Nuclepore filters on. It really reduced static. The only downside was the
half-life of a few months that meant after a year or so you had to buy
another. We also had Zero-Stat gun, but that wasn't used as it simply didn't
work. The Nuclepore device seems to have been dropped now, but you can buy a
brush that has Polonium-210 trapped within the bristles that should work on
similar lines (NRD - StaticMaster Anti-Static Brush - 1" or 3") :

http://www.optcorp.com/product.aspx?pid=1818&kw=staticmaster&st=2

I know that photographers use them to reduce static on film. I've never
tried this specific brush though as I have now moved out of this field. Our
histologist used something similar on microtome surfaces when sectioning I
remember.

Also have a look at ionisers like :
http://www.etherton.co.uk/index.html
http://www.2spi.com/catalog/photo/antistat.html

Alternatively you can spray a conducting coating onto the plastic to
eliminate static (e.g. the Duron anti-static spray sold by Agar Scientific
that "even allows uncoated samples to be viewed under 'low resolution' in an
SEM").

There's also the anti-static wrist-strap (or just touching the metal earth
of a plugged in PC case or wall socket screw head). Plus breathing on the
plastic really removes the static (or earthing the material if it conducts),
but that suits plastic record player Perspex covers more than scientific
specimens. Keeping the room %RH constant and not going too low will also
help. Some labs have a full antistatic grounding system with furniture, mats
etc.. all 'grounded' with controlled %RH and temperatures, but we got by
with antistatic mats, higher humidity and the polonium-210. We avoided using
gloves as much as possible, using fine metal forceps for handling filters
(I don't think we bothered with 'antistatic' forceps). Antistatic gloves
weren't much use as we needed disposable ones. We used very thin sensitive
clear plastic gloves that were probably something like Polyethylene,
although they probably did have some static problems. Our Nuclepore filter
samples were generally glassfibres recovered from lungs (fibre durability +
toxicology studies) or aerosol particles, and obviously when using aqueous
samples static was only a problem with the clean filter prior to filtration
or after drying.

Regards

Keith

----------------------------------------------------------
Dr Keith J Morris
Imaging Facilities Manager
Cell Biology Division
Institute of Ophthalmology
University College London
11-43 Bath Street
London EC1V 9EL

Tel: 020 7608 4050
Fax: 020 7608 4034
email: keith.morris-at-ucl.ac.uk



----- Original Message -----
X-from: {TindallR-at-missouri.edu}
To: {keith.morris-at-ucl.ac.uk}
Sent: Thursday, January 12, 2006 11:04 PM

I'll send you a PDF of the salary survey article that appeared in
Microscopy Today last year. Also, if you are going to be a microscopist
(congratulations!) you should sign up for a free subscription to
Microscopy Today at http://www.microscopy-today.com.

Ron Anderson, Editor

dustin.adams-at-xerox.com wrote:

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From: TindallR-at-missouri.edu
Date: Fri, 13 Jan 2006 08:50:02 -0600
Subject: [Microscopy] Static!

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Hi all,

Thanks for all the suggestions on static control! I learned about some
new things to try for general use. I had forgotten about the polonium
strip StaticMaster brushes from my days locked in a photo darkroom, but
this may be a good alternative to these outrageously overpriced Zero
Stat guns that don't work nearly as well as they did a few years ago.

Those StaticMaster brushes used to strike fear into us when we read the
disposal instructions, which were something like "seal this strip inside
a deep, deep mine and don't go anywhere near it for 5000 years",
although we could use them in the darkroom with no special protection.
They did work, though, except you couldn't spark your co-workers with
them and make them squeal.

By the way, our most common use for these guns is somewhat trivial:
getting grids down off the lid of the petri dish, where static
electricity glues them, especially in winter. (Tamara, I agree with you
about the static in New Mexico---I still have scars from getting zapped
every time I'd get out of my car in Las Cruces!). We use the Diatome
de-ionizer for microtomy, which works well.

Thanks again!

Randy

Randy Tindall
EM Specialist
Electron Microscopy Core Facility---We Do Small Well!
W122 Veterinary Medicine
University of Missouri
Columbia, MO 65211
Tel: (573) 882-8304
Fax: (573) 884-2227
Email: tindallr-at-missouri.edu
Web: http://www.emc.missouri.edu







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From: paul_hazelton-at-umanitoba.ca
Date: Fri, 13 Jan 2006 09:59:12 -0600
Subject: [Microscopy] Re: Static!

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Randy

You want static??!!! Try Manitoba in February. When it's -30C there's
no such thing as moisture in the air. But then, not even Phil was
willing to try Manitoba in June/July, even for real beer ;-) .

Great time to make formvar grids, though.

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
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From: paul_hazelton-at-umanitoba.ca
Date: Fri, 13 Jan 2006 09:59:42 -0600
Subject: [Microscopy] Re: bacterial permeabilization - further

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Tom

Definition of milli-second is the amount of time between hitting send
and realizing there is an error, da? Of course I meant phospholipid,
not lipoprotein when mentioning the outer and cytoplasmic membranes.
Mark it up to trying to do e-mail when the brain is turned off. Should
Ron Anderson decide that this is a train worth following for Microscopy
Today and want to use my response to you, perhaps he could make the
correction, please??!!

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: paul_hazelton-at-umanitoba.ca
Date: Fri, 13 Jan 2006 10:12:07 -0600
Subject: [Microscopy] Re: bacterial permeabilization

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Tom

Very interesting question. Made more interesting to me by the fact that I was discussing this same topic this afternoon, only more generically, and certainly not in reference to H. influenza. The problem you are faced with is bacterial structure. H. influenza is a Gram negative microorganism. Gram negative cells have two lipoprotein layers - the outer and cytoplasmic membranes. In addition, you have a layer of peptidoglycan between the outer and cytoplasmic membranes. To make it even more fun, there is frequently a lypopolysaccharide layer around the whole thing. You have to penetrate the LPS, permeabilize the outer membrane, traverse the periplasmic space (the volume between the outer and cytoplasmic membranes), cross the peptidoglycan layer and then penetrate the inner membrane, all with gentle treatment to allow the membranes to reorganize after removal of the detergent you use for permeabilization. What it adds up to is that permeabilization will be difficult at best, and most likely impossible. Probably the only way to get something reliably into the cytoplasm would be by active transport.

Having said that, knowing what I do about the structure of prokaryotic cells, I confess to having tried to permeabilize and do pre-embedding labeling on E. coli expressing a protein of interest (another gram negative bacterium). Didn't expect it, but tried anyway. Hey, dogma schmogma, give it a try, right? That's how the serendipitous findings occur. Anyway, used triton X, used saponin. Ugly. Did not look good. No luck either way. So logic and dogma were right this time.

At the same time I did LR white embedding on cells from the same broth culture. Included osmium fixation. Went back with metaperiodate followed by hydrogen peroxide etching and then did indirect IEM with 12nm gold for the label. Beautiful preservation. Got highly significant labeling of the expressed protein in the E. coli and in the wild type bacterium from which the protein of interest had been cloned. In fact, the protein of interest was hypothesized to be membrane inserted on the basis of amino acid sequence and the labeling of the wild type cells was primarily associated with the cytoplasmic membrane. In short, I did minor modification to the standard LR white embedding protocol and it worked great. I would give that a try.

I would like to give a reference for the work, but we are still waiting for the student to finish his thesis, not to mention the paper. And he has already left for his post-doc. If you have any questions, call. I would be glad to discuss the procedure I used further.

Paul




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From: bfoster-at-mme1.com
Date: Fri, 13 Jan 2006 10:57:55 -0600
Subject: [Microscopy] Re: AskAMicroscopist: SEM microscopist salaries

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

Let's try this with my fingers working ...

Hey! I would've been happy to go to Manitoba, even for real beer. Just kind of hard when I was busy fending off predatory administraitors.
Gave up on that and just moved.

Phil

Philip Oshel
Microscopy Facility Supervisor
Department of Biology
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576


Hi, Dustin

We did a salary survey for the society in conjunction with Microscopy Today late last year. Contact Ron Anderson for details. (see email above).

Hope this is helpful,
Barbara Foster

Microscopy/Microscopy Education
313 S Jupiter Rd, Suite 100
Allen, TX 75002
P: 972-954-8011
W: www.MicroscopyEducation.com

P. S.
Need a good general reference or light microscopy text for the Spring semester? Call us today to learn more about "Optimizing LIght Microscopy". Copies still available through MME... even for class-room lots ... and we give quantity discounts. Call Ken Piel at (972)954-8011.



At 08:25 PM 1/12/2006, dustin.adams-at-xerox.com wrote:



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From: bfoster-at-mme1.com
Date: Fri, 13 Jan 2006 11:03:21 -0600
Subject: [Microscopy] Re: AskAMicroscopist: SEM microscopist salaries

Contents Retrieved from Microscopy Listserver Archives
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At 08:25 PM 1/12/2006, dustin.adams-at-xerox.com wrote:



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From: phillipst-at-missouri.edu
Date: Fri, 13 Jan 2006 11:52:57 -0600
Subject: [Microscopy] Hi end digital camera

Contents Retrieved from Microscopy Listserver Archives
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I am looking at getting a deeply cooled (-30 C) high sensitivity, high
resolution digital camera. I am considering the following cameras :
Hamamatsu Orca-AG
Retiga SRV
Photometrics CoolSnap HQ or K4

Any comments (public or private) on your experiences would be appreciated.

One source has told me that they see little practical difference in the
Orca (cooled to -30) vs the Retiga EXi (cooled to 25 below ambient).
Comments about this statement are also welcome.

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



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From: eschumacher-at-mccrone.com
Date: Fri, 13 Jan 2006 12:14:59 -0600
Subject: [Microscopy] Open Position: Electron Microscopy Technician

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

McCrone Associates, Inc., an innovative modern analytical and research
laboratory, seeks a highly motivated electron microscopy technician with
training in ultramicrotomy for materials science. The successful
candidate will support our Electron Optics Group in sample preparation
for SEM and TEM, as well as instrument maintenance. Ability to carry
out routine SEM and TEM imaging and x-ray analysis is also desired. For
more information on The McCrone Group, please visit www.mccrone.com.

The ideal candidate will have a B.S. degree with coursework in electron
microscopy and microtomy, or comparable experience. Compensation is
commensurate with education, experience and responsibilities.

Applicant selected must be a U.S. Citizen, will be subject to a
government security investigation, and must meet eligibility
requirements for access to classified information. McCrone is an Equal
Opportunity Employer.

If you have microtomy experience and are looking for an interesting and
unique career opportunity in materials analysis, send your resume with a
letter describing your experience and interests to:

Careers
McCrone Associates, Inc.
850 Pasquinelli Drive
Westmont, Illinois 60559

FAX: 630-887-7417 - Attn: Human Resources
E-mail: careers-at-mccrone.com (e-mail is preferred)



Elaine Schumacher
Senior Research Scientist
McCrone Associates, Inc.
850 Pasquinelli Drive
Westmont, IL 60559-5539 USA
630-887-7100 (tel)
630-887-7417 (fax)
E-mail: eschumacher-at-mccrone.com
Web Site: www.mccrone.com



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From: rbeavers-at-mail.smu.edu
Date: Fri, 13 Jan 2006 15:12:52 -0600
Subject: [Microscopy] Lab Facilities Temperature

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Hi

What you are seeing is the projected image of the hot filament (light).
This means that although you have a hole down the column the electron gun is
out of alignment. May I suggest the following.

At no more than 60kV switch off all the lenses and look for a very bright
electron beam by adjusting the gun alignment controls. Once you find the
beam switch each lens on one at a time, stepping back if the beam
disappears.

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 7802 966067 Web www.emcourses.com

----- Original Message -----
X-from: {aetmicro-at-optonline.net}
To: {protrain-at-emcourses.com}
Sent: Friday, January 13, 2006 2:22 AM

Group,

Our University in an effort of conservation of energy is making changes as to the HVAC conditions they can provide us. Requests as to equipment shuts downs during non use hours, shuting off computers every time not in use, and increasing lab temperatures beyound 78F are being asked for. Looking for documented case studies pro or con and anyones experience they would be willing to share offline on this issue. Would also like to hear from vendors as to desired environments for SEM and related equipment as well as stored chemicals.

Thanks

Roy Beavers

Southern Methodist University
Department of Geological Sciences
P.O. Box 750395
Dallas, TX 75275
Voice: 214-768-2756
Fax: 214-768-2701
Email: rbeavers-at-smu.edu



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From: mswift-at-bunham.org
Date: Sat, 14 Jan 2006 07:34:52 -0600
Subject: [Microscopy] AskAMicroscopist: Tecnai 12 Focus Substates

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 (mswift-at-bunham.org) from http://microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Friday, January 13, 2006 at 18:18:43
---------------------------------------------------------------------------

Email: mswift-at-bunham.org
Name: Mark Swift

Organization: Burnham Institute

Education: Graduate College

Location: San Diego, CA

Question: We have a Tecnai 12 from FEI, and we would like to know: How do we ensure that in Low Dose our Focus Substates 1 and 2 are on the tilt axis when Alpha angle is not 0 degrees?

Thanks,

Mark

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

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From: lemaster-at-ppg.com
Date: Mon, 16 Jan 2006 09:31:25 -0600
Subject: [Microscopy] SEM advice on low cost polisher

Contents Retrieved from Microscopy Listserver Archives
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I am seeking advice on obtaining a low cost polisher for SEM sample preparation. The samples will be coated thin metal cross-sections embedded in epoxy. The cross-sections will be analyzed by SEM to determine coating thickness. I have a diamond wet saw but I need a polisher but have very little funds available. I found a used Buehler Minimet polisher in my price range ~$1K. I am not familiar with this unit. I have used bigger units like the Buehler Powerpro and the Leco SS-2000. Is the difference mainly a matter of speed and sample size? I plan on doing only about one sample a week. Any advice will be appreciated.
Thanks,

Scott LeMaster
PPG Industries Inc.
lemaster-at-ppg.com



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From: r-holdford-at-ti.com
Date: Mon, 16 Jan 2006 10:43:49 -0600
Subject: [Microscopy] Re: SEM advice on low cost polisher

Contents Retrieved from Microscopy Listserver Archives
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Scott: on the MiniMet, the sample moves and the polishing media stays
still. This tool can take up to a 2 inch potted sample and uses 4-inch
PSA grinding/polishing media on glass discs. It was designed to do
rough and fine polish and it does that pretty well. On this tool, you
drill a small blind hole in the top of your sample. This is where a
spring-loaded arm fits to move the sample in an random orbital motion.
You have some control over the pressure of the sample into the media and
the speed of rotation. It can run unattended. This tool might be just
what you need for one sample a week, if you can cut almost to the area
of interest and then polish the rest of the way. On used tools, the
spring force is usually the first thing to go due to rust, so be aware.

lemaster-at-ppg.com wrote:

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--
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
Becky Holdford (r-holdford-at-ti.com)
972-995-2360
972-648-8743 (pager)
SC Packaging FA Development
Texas Instruments, Inc.
Dallas, TX
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~


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From: eschumacher-at-mccrone.com
Date: Mon, 16 Jan 2006 11:56:34 -0600
Subject: [Microscopy] Short Course Announcement: Scanning Electron Microscopy

Contents Retrieved from Microscopy Listserver Archives
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Fellow Listers,

The College of Microscopy will be offering a short course, Scanning
Electron Microscopy, March 27-31, 2006, at our Westmont Facility. In
addition to lectures, the course emphasizes hands-on training using five
scanning electron microscopes and electron microprobe analyzers, and
gives students the opportunity to work on their own samples.
For further details and registration information, please follow the link
below.

www.collegeofmicroscopy.com



Elaine Schumacher
Senior Research Scientist
McCrone Associates, Inc.
850 Pasquinelli Drive
Westmont, IL 60559-5539 USA
630-887-7100 (tel)
630-887-7417 (fax)
E-mail: eschumacher-at-mccrone.com
Web Site: www.mccrone.com



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From: henriks-at-southbaytech.com
Date: Mon, 16 Jan 2006 13:06:36 -0600
Subject: [Microscopy] Re: SEM advice on low cost polisher

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

Dear Scott:

DISCLAIMER: South Bay Technology produces equipment and supplies as
described below and, therefore, has a vested interest in promoting their
use.

If you are mounting the sample in AcryliMet™ cold mount or something
similar and can hold your sample by hand, then something like our Model
900 Grinder/Polisher would be ideal. A new Model 900 falls into about
the same price range as the used Minimet you referred to. This system is
often used for low volume polishing requirements - although usually for
higher volume than just 1 sample per week. If you have unencapsulated
samples or simply want more control over the polishing process, you can
use the Model 900 together with something like our Tripod Polisher®
Cross Section Polisher, our BiPod™ Cross Section Polisher or one of our
other lapping and polishing fixtures. You can find information on the
Model 900 Polisher at
http://southbaytech.com/products_index.cfm?main_action=product_detail&ProductID=42.

If you do end up with the Minimet Polisher, I do have a large lot of
Minimet Consumables available at a huge discount. If you have an
interest in those, please let me know and I will send you the listing.

Best regards-

David


lemaster-at-ppg.com wrote:

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South Bay Technology, Inc.
1120 Via Callejon
San Clemente, CA 92673 USA

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email: henriks-at-southbaytech.com

Celebrating 40 years of providing Materials Processing Solutions for Metallogaphy, Crystallography and Electron Microscopy.

Please visit us online at www.southbaytech.com.

The information contained in this message and any attachments is privileged and confidential. This message is intended for the individual or entity addressed. If you are not the intended recipient, please do not read, copy or disclose this communication. Notify the sender of the mistake by calling +1-949-492-2600 and delete this message from your system.





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From: bigelow-at-engin.umich.edu
Date: Mon, 16 Jan 2006 14:34:38 -0600
Subject: [Microscopy] RE: Starting Ion Pumps without cooling water

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


I just returned from a trip out of the country, and found an inquiry
about starting up ion pumps in among 1200 other assorted e-mail
messages (mostly spam), and so I apologize for being so delayed in
answering.

I don't know the exact details of the arrangement of the vacuum
system on a JEOL 2010, but in general there should be no requirement
for having cooling water running while starting up or running a
sputter ion pump. In fact, one of the principle advantages of these
pumps is the fact that they do not need cooling of any sort for
normal operation.

What you would need to do is to set the valves in the vacuum system
so that the backing pump evacuates the region of the vacuum system
served by the ion pumps, then evacuate this region to a vacuum in the
low 10-3 torr range. Then, follow the start-up procedure recommended
for your ion pumps. You want to minimize the time the backing pumps
are operated in the low end of this pressure range to reduce the
possibility of contaminating the system by back streaming of oil from
them, otherwise, there should be no problem starting up the ion
pumps. As soon as they do start you want to close whatever valves
are available to isolate the part of the system they serve from the
part evacuated by the other pumps. Once the supply of cooling water
is restored, use the diffusion pump to evacuate the rest of the
system to an operating level, and then open the valves to the
ion-pumped region and resume normal operation.

If you can get hold of a copy of my book on Vacuum Methods in
Electron Microscopy (ISBN 1 85578 053 4) you can find the matter of
backstreaming from rotary vane pumps discussed in Section 4.1.5 on p.
144. The general operating characteristics of sputter ion pumps is
discussed in Sections 7.1.6, p 290 and 7.5, p. 322. I believe the
operating procedure for the vacuum system described in Section 9.3 on
p. 392 is very similar to that for the 2010.

Good luck,
Wil Bigelow

--
Wilbur C. Bigelow, Professor Emeritus
Materials Sci. & Engr., Univ. of Michigan
Ann Arbor, Michigan 48109-2136
e-mail: bigelow-at-engin.umich.edu;
Fx:734-763-4788; Ph:734-662-5237
Address mail to: 1136 Mixtwood Rd.
Ann Arbor, MI 48103-3035

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From: gary-at-gaugler.com
Date: Mon, 16 Jan 2006 15:40:10 -0600
Subject: [Microscopy] Starting Ion Pumps without cooling water

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


I too am not aware of any water cooling for ion pumps.
when first pumping, they do get warm. But their controller
will shut them off for a cool down.

The main problem I have had in the past is getting them to
start pumping. Initially, no pump current. This is fixed
by heating the pump with a hair dryer. Then they fire and
that is that. This is typical of the little 1-5L/m gun
chamber pumps for LaB6. The larger 30L/m pumps don't seem
to have this problem. This was with Varian pumps.

gary g.



At 12:53 PM 1/16/2006, you wrote:



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From: tom-at-tomkaye.com
Date: Mon, 16 Jan 2006 20:44:12 -0600
Subject: [Microscopy] Question about vacuum gauge on JEOL T300

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

Hello All,

I have a multi-function gauge on my T300 that reads from 0-1. It is used
with a rotating switch to measure all the voltages etc in the system. When
its at full vacuum it reads 0.42 and the manual says it should be 6v which I
assume is 0.6. I was wondering if anyone out there knows what this
translates to in terms of Torr? Please reply off list if this is not of
general interest.

Thanks in advance!

Tom Kaye


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5, 20 -- Subject: Question about vacuum gauge on JEOL T300
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From: cumings-at-umd.edu
Date: Mon, 16 Jan 2006 21:05:32 -0600
Subject: [Microscopy] viaWWW: Job Opening

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

This Question/Comment was submitted to the Microscopy Listserver
using the WWW based Form at http://www.microscopy.com/MLFormMail.html
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Email: cumings-at-umd.edu
Name: John Cumings

Organization: University of Maryland

Title-Subject: [Filtered] Job Opening

Question: Dear Microscopy Community,

I would like to bring your attention to an open position in electron microscopy as the lab manager of a newly-created facility here at the University of Maryland. The position is part of the growing nanoscience center on campus, the Maryland Center for Integrated Nano Science and Engineering (M-CINSE).

More information can be found at
http://mse.umd.edu/dept/positions/LabManagerUniversityMicroscopyFacility.htm

and applications can be submitted at
https://apra.umd.edu/search.jsp?ID=ENMA000003

Please note that the "best consideration" deadline of Jan. 15th has just passed, so please apply soon if you are interested.

Best regards,

-John Cumings

--
John Cumings
cumings-at-umd.edu
Assistant Professor
Department of Materials Science and Engineering
University of Maryland
College Park, MD 20742-2115

office (301) 405-0789 (1246 Kim Building)
fax (301) 314-8164
--


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From: nrvrctr-at-adelphia.net
Date: Mon, 16 Jan 2006 21:06:12 -0600
Subject: [Microscopy] viaWWW: Test engineer in Chicago

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

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Email: nrvrctr-at-adelphia.net
Name: Ed Beckett

Organization: MRI Virginia

Title-Subject: [Filtered] SEM

Question: I am searching for a Test engineer in Chicago with experience in SEM as well as other electrical and test abilities. Salary is about $60K. Contact me if you are aware of anyone or self interested. Thanks Ed Beckett
540-980-3100

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

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From: yezer-at-cc.hut.fi
Date: Tue, 17 Jan 2006 02:22:25 -0600
Subject: [Microscopy] Magnetic contrast with 4QBSED

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

Hi,


I am interested to study magnetic structure (type II magnetic contrast) by
SEM. Our SEM equipped with a Four Quadrant Backscattered Electron Detector. I
will appreciate if you have experience using this type of detector for
magnetic domain structure to guide us with the recommended working parameters
as combinations of quadrant setting, working distance, tilt and etc.

Thank you in advanced,

Yossi.

==============================Original Headers==============================
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From: fonda-at-anvil.nrl.navy.mil
Date: Tue, 17 Jan 2006 06:21:11 -0600
Subject: [Microscopy] Postdoctoral Fellowship opening at NRL

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

The Naval Research Laboratory has a current opening for a
Postdoctoral Fellow in the Joining and Transformations Section. The
ideal candidate would have a strong background in both electron
microscopy and materials science. The successful candidate will
study the transformations that occur during joining processes and
relate the microstructural features to observed mechanical
properties. Current research interests center on the precipitation,
dynamic recrystallization, and texture evolution that occur during
the friction stir welding process.

The Naval Research Laboratory has an excellent array of microscopy
facilities available. There are three transmission electron
microscopes (a Phillips CM-30 analytical TEM, a Hitachi H-9000
high-resolution TEM, and a new, state-of-the-art JEOL 2200 energy
filtered STEM/TEM), two scanning electron microscopes (a Leo 1550 SEM
with electron backscattered diffraction analysis and energy
dispersive spectrometry and a Hitachi FEG-SEM), and a dual-beam
focused ion beam with EBSD that are available for use by the
successful candidate. Additional facilities that may be used include
a quenching and deformation dilatometer, a Gleeble thermomechanical
simulator, x-ray diffractometers, and an automated microhardness
tester.

Please note that this position is only open to US citizens or Green
Card holders.

Please contact me by e-mail (Fonda-at-anvil.nrl.navy.mil) or phone
(202-767-2622) for further details about this research program.

Sinceerely,

Richard Fonda
--
________________________________________________________________
Richard W. Fonda Naval Research Laboratory
(202) 767-2622 Code 6324
(202) 767-2623 fax Washington DC 20375
________________________________________________________________

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From: lleroux-at-csir.co.za
Date: Tue, 17 Jan 2006 07:34:30 -0600
Subject: [Microscopy] viaWWW: model S150B Sputter Coater

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Email: lleroux-at-csir.co.za
Name: Lukas le Roux

Organization: CSIR

Title-Subject: [Filtered] Edwards Sputter Coater

Question: I am trying to find an instruction manual for a model S150B Sputter Coater. Is there a website from where I can download it?

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From: emlabservices-at-cox.net
Date: Tue, 17 Jan 2006 07:35:51 -0600
Subject: [Microscopy] TEM Room Survey Company

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Hello All,

Is there a company that can provide room survey services in the Dallas,
Texas area? Environmental testing for mechanical vibration and stray EMF
parameters are desired.

Please respond directly off list.

Bob Roberts
EM Lab Services, Inc.
449 NW 62nd St.
Topeka, Kansas 66617-1780
785.246.1232 voice
785.246.0168 fax
www.emlabservices.com



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From: Gareth.Morgan-at-ki.se
Date: Tue, 17 Jan 2006 10:10:43 -0600
Subject: [Microscopy] Image analysis

Contents Retrieved from Microscopy Listserver Archives
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Hi

Can anyone out there help a colleague?

"I am discussing proposals to install an imaging
system here. It is one that can capture both FISH
and brown/ H&E stains and then allow an operator,
using selected software, to analyse them. The
favoured system at present is the Applied Imaging
one as it is now undergoing a major facelift.

I am just wondering if there are any other
systems on the market right now that can load
up 50 slides automatically overnight, scan &
store so that an operator can then analyse the
images captured the following morning. None of
the other major suppliers here in the UK, such as
ChromaVision, Meta-systems & Aperio can offer
both overnight loading and scanning of both FISH
and brown/ blue & pink stains at present."


Med vänliga hälsningar/With best regards

Gareth

http://www.ki.se/biomedlab
e-mail Gareth.Morgan-at-ki.se

Tel +46 8 5858 1038
Fax +46 8 5858 7730

Gareth Morgan MPhil MSc FIBMS,
Department of Histo/cytopathology, Laboratory Medicine (Labmed),
Karolinska Institute,
Karolinska University Hospital at Huddinge, F46
SE 141 86 Stockholm
Sweden

OBS! Besöksadress: F-Huset, Forskningsgatan 2
F52, Rum 2.10. Laboratoriet för klinisk patologi och cytologi.

NB! Visiting address: Building F, Research
Corridor 2 F52, Room 2.10. Clinical Histo- and Cytopathology Laboratory.



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From: marcia_pitzenberger-at-merck.com
Date: Tue, 17 Jan 2006 11:12:37 -0600
Subject: [Microscopy] TEM - Electron Beam Clearing

Contents Retrieved from Microscopy Listserver Archives
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TEM User's -


We're having trouble with bright spots appearing on our tissue when viewing
our grids in our new TEM. These spots may be described as bleaching,
etching, burning or clearing. The bright spots appears after using high
magnification to view an area or to focus. When we go back to a low
magnification, we see the bright area.

This started happening with the installation of a new TEM. We haven't
changed processing procedures and we are experienced instrument operators.
The service engineer has determined that the TEM is working properly. The
problem is not due to prolonged exposure to the electron beam. The spots
occur within seconds. The filament is positioned correctly. Often there is
variability in the intensity of the spotting even on the same grid. In
other words, sometime the spot is very apparent and distinctive and other
times it's more diffuse. We've tried using liquid nitrogen to improve the
vacuum and adding a second 100% resin infiltration step to ensure removal of
water from the specimen. The problem occurs with cells and tissues
embedded in epoxy. The vendor has suggested working at 120kV and we're
starting to do that now. It doesn't entirely correct the problem, but the
spots are less focal.

Has anyone worked with a particular epoxy that stands up well or better than
others under the electron beam? You can reply off list. Has anyone had
this problem and been able find a solution?

Thank you in advance for your time

Marcia



Marcia Pitzenberger
Pathology Laboratories
Merck & Co., Inc.
P.O. Box 4, WP45-104
West Point, PA 19486-0004
Tel 215 652-9767
Fax 215 652-7758
marcia_pitzenberger-at-merck.com





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From: tivol-at-caltech.edu
Date: Tue, 17 Jan 2006 12:14:07 -0600
Subject: [Microscopy] Re: AskAMicroscopist: Tecnai 12 Focus Substates

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On Jan 14, 2006, at 5:35 AM, mswift-at-bunham.org wrote:

} Question: We have a Tecnai 12 from FEI, and we would like to know: How
} do we ensure that in Low Dose our Focus Substates 1 and 2 are on the
} tilt axis when Alpha angle is not 0 degrees?
}
Dear Mark,
If you set the focus angles for the substates to 0 and 180 degrees,
the displacements will be along the tilt axis. You can verify this by
burning a small hole in the ice on a cryogrid in each of the focus
substates, then going to a lower mag, where the holes are both visible
in the exposure state, and tilting the grid.
Yours,
Bill Tivol, PhD
EM Scientist and Manager
Cryo-Electron 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: ron.doole-at-materials.ox.ac.uk
Date: Tue, 17 Jan 2006 12:26:02 -0600
Subject: [Microscopy] Re: TEM - Electron Beam Clearing

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Hi Marcia,

I speak from a position of ignorance of your type of specimen, however I have
picked up on 'This started happening with the installation of a new TEM'.

Are you using a much higher beam current at high mags than previously? Modern
TEMs have much better condenser optics than the older ranges available in the
70s and 80s with less use of apertures to reduce spot size and better use of
lenses. Have you changed from tungsten to LaB6 or FEG?

These changes can result in more electrons getting to the specimen. Can you
compare the beam current that you use at high mag on the two TEMs? If you know
typical exposure times of the two instruments do you have to spread the beam
more on the new instument to get the same exposure time? Try using a smaller
spot size or smaller condenser aperture and see if the effect improves.

Good luck,
Ron

In message {200601171752.k0HHqH72018071-at-ns.microscopy.com}
marcia_pitzenberger-at-merck.com writes:
}
}
}
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
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} TEM User's -
}
}
} We're having trouble with bright spots appearing on our tissue when viewing
} our grids in our new TEM. These spots may be described as bleaching,
} etching, burning or clearing. The bright spots appears after using high
} magnification to view an area or to focus. When we go back to a low
} magnification, we see the bright area.
}
} This started happening with the installation of a new TEM. We haven't
} changed processing procedures and we are experienced instrument operators.
} The service engineer has determined that the TEM is working properly. The
} problem is not due to prolonged exposure to the electron beam. The spots
} occur within seconds. The filament is positioned correctly. Often there is
} variability in the intensity of the spotting even on the same grid. In
} other words, sometime the spot is very apparent and distinctive and other
} times it's more diffuse. We've tried using liquid nitrogen to improve the
} vacuum and adding a second 100% resin infiltration step to ensure removal of
} water from the specimen. The problem occurs with cells and tissues
} embedded in epoxy. The vendor has suggested working at 120kV and we're
} starting to do that now. It doesn't entirely correct the problem, but the
} spots are less focal.
}
} Has anyone worked with a particular epoxy that stands up well or better than
} others under the electron beam? You can reply off list. Has anyone had
} this problem and been able find a solution?
}
} Thank you in advance for your time
}
} Marcia
}
}
}
} Marcia Pitzenberger
} Pathology Laboratories
} Merck & Co., Inc.
} P.O. Box 4, WP45-104
} West Point, PA 19486-0004
} Tel 215 652-9767
} Fax 215 652-7758
} marcia_pitzenberger-at-merck.com
}
}
}
}
}
} -----------------------------------------------------------------------------
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Banyu) that may be confidential, proprietary copyrighted and/or legally
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}

--
Mr. Ron Doole Department of Materials
Senior Instrumentation Engineer. University of Oxford.
Phone +44 (0) 1865 273701 Parks Road. Oxford. OX1 3PH
Fax +44 (0) 1865 283333 ron.doole-at-materials.ox.ac.uk

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From: john.mardinly-at-intel.com
Date: Tue, 17 Jan 2006 15:16:14 -0600
Subject: [Microscopy] Starting Ion Pumps without cooling water

Contents Retrieved from Microscopy Listserver Archives
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Gary;
Hitting ion pumps with a hammer also gets them started. The
hammer is also very good at knocking loose whiskers that form in the ion
pump.

John Mardinly
Intel

The opinion of this author does not necessarily represent the opinion of
Intel Corporation.

-----Original Message-----
X-from: gary-at-gaugler.com [mailto:gary-at-gaugler.com]
Sent: Monday, January 16, 2006 1:41 PM
To: Mardinly, John


I too am not aware of any water cooling for ion pumps.
when first pumping, they do get warm. But their controller
will shut them off for a cool down.

The main problem I have had in the past is getting them to
start pumping. Initially, no pump current. This is fixed
by heating the pump with a hair dryer. Then they fire and
that is that. This is typical of the little 1-5L/m gun
chamber pumps for LaB6. The larger 30L/m pumps don't seem
to have this problem. This was with Varian pumps.

gary g.



At 12:53 PM 1/16/2006, you wrote:



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From: larry-at-cymru.freewire.co.uk
Date: Tue, 17 Jan 2006 15:45:46 -0600
Subject: [Microscopy] Re: SEM advice on low cost polisher

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Several glass plates ~5 mm thick and 15 cm square. Polishing clothes
stuck to the glass plates. Glass plates inside large tray. Water
spray to lubricate. Cost is almost nothing apart from the time.
Depends how large the sample is and what finish you are working from.
Fine grinding can be done by using aumina directly onto the glass.
It's the way I used to prepare 3 mm steel discs for TEM, before a
final electropolish. A 3 mm stainless steel disc takes ~15 mins to
polish by hand from the alumina grinding.

--
Larry Stoter

PLEASE NOTE
1. Any mail other than plain text will be automatically deleted.
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From: gary-at-gaugler.com
Date: Tue, 17 Jan 2006 17:13:09 -0600
Subject: [Microscopy] Starting Ion Pumps without cooling water

Contents Retrieved from Microscopy Listserver Archives
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Oh yeah. I forgot to mention that trick too. I usually
use the handle end of a screwdriver. This also works
for cold cathode sensors too.

gary g.


At 01:33 PM 1/17/2006, you wrote:



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From: dsams-at-schaferlabs.com
Date: Wed, 18 Jan 2006 13:21:03 -0600
Subject: [Microscopy] Thermal Ionization Mass Spectrometry Senior Engineer / Scientist Job Opening at Schafer Corporation

Contents Retrieved from Microscopy Listserver Archives
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With some degree of simplification- low signal electronic imaging is a race
between noise accumulation and signal accumulation.

Some of the noise sources in the CCD are:

a) thermal noise (reduced by factor of 2 for every 6 to 7 degrees C
temperature drop for silicone CCD), mostly correctable by dark frame
subtraction;
b) readout (pixel clock switching) noise also know as bias noise, somewhat
reduced by cooling, correctable by bias frame subtraction;
c) random (shot) noise - a random part of thermal noise (a) which can not be
corrected by dark frame subtraction- this noise gets worse with pixels size
shrinking (the larger the pixels the less random this value is)- a crude
comparison is a mechanical vibration dumping by increasing weight
(anti-vibration platform).

Assuming your signal source is set (optics, specimen, illumination):

a) the colder the sensor- the better S/N ratio is- noise accumulation slows
down, while signal stays the same;
b) with 30 deg. C difference noise will be different by factor of 32 (30deg.
= 6deg.*5; noise drops twice per every 6 degrees, thus 2 power 5 = 32 which
is a dramatic noise reduction, but noticeable only if light is dim and
exposures are long.
c) the larger the pixels- the better S/N ratio is- at the expense of spatial
resolution- it could pay to have more smaller pixels and use binning instead
when needed.

Type of sensor:

a) definitely CCD over CMOS;
b) probably full frame CCD over interline CCD, (full frame has much better
S/N ratio for a given pixel size), but interline CCD could be more
convenient due to potentially faster frame rate, exposure time allowing;
c) if exposure time must be long, interline CCD looses it's potential speed
advantage, and it is always noisier than full frame one;
d) interline CCD has typically 50% of it's light receiving area occupied by
transfer gates, meaning that only half of it is light sensitive- this can be
improved by micro lens array - most interline CCDs are available in such
configuration), but sensitive area of the pixel is still half of what it
could be in full frame CCD with same pixel size, which means higher noise .

Interline CCDs can use frame integration with less or no cooling and achieve
decent noise reduction, but this is always inferior to deeply cooled full
frame CCD in single frame (long exposure) integration mode, with respect to
S/N.

I deliberately used S/N (signal to noise) ratio instead just "noise",
because it is precisely ratio that counts. If signal is strong (bright
light), then it accumulates much faster than noise. Then exposures are
short, and no CCD cooling needed. There is no straight answer without
knowing the numbers for a given application.

A specific application could be best served by either a CMOS, or a full
frame CCD, or interline CCD sensor, cooled or not- many more variables must
be taken into account.

Vitaly Feingold
SIA
2773 Heath Line
Duluth GA 30096
Ph. 770-232-7785
Fax 770-232-1791
www.sia-cam.com

----- Original Message -----
X-from: {phillipst-at-missouri.edu}
To: {vitalylazar-at-att.net}
Sent: Friday, January 13, 2006 12:54 PM

Overview: Schafer Vallecitos Laboratory is seeking a Senior Engineer
/Scientist to add to our mass spectrometry group. SVL performs materials
characterization and related analytical services on commercial and
government contracts. The activities of the group include chemical and
elemental analysis of materials on a production basis, maintenance of
several mass spectrometers and ancillary equipment, development of new or
improved MS analysis techniques, and quality assurance of analytical data.
Schafer is a technically strong firm with a reputation for quality and
integrity. Schafer's reputation is a direct result of our dedicated,
motivated, talented, and creative staff that is responsible for developing
our outstanding business relationships with our customers. Our technical
capabilities are vast and growing to provide innovations for the future.

Responsibilities: This position requires an individual who has proven
technical abilities to function as an instrument designer, with emphasis on
ion optics, electronics, and computer automation. Commercial design
experience would be a plus, as the instrument should be maintainable and
replicable, not a one-of-a-kind that requires a large staff to maintain. The
ideal candidate must have a strong background in materials science or
engineering, or a related field of physics, geology, chemistry, or
metrology. Experience in design and operation of mass spectrometers,
particularly TIMS, and other analytical instruments, is highly recommended.
The candidate should have competence in vacuum technology, electronics,
mechanical design, ion optics, and project administration.

Qualifications: Experience with analytical instrument control computer
hardware and software is required. The candidate must be able to operate
independently with occasional supervision.

Other qualifications include:

. Bachelor's degree in physical science or engineering.
. Minimum 10 years of technical experience.
. Demonstrated ability to solve technical problems.
. Expertise in data evaluation and quality control.
. Proven ability to write clear scientific reports and proposals.
. Must be a US citizen with the ability to obtain government security
clearance.

Schafer offers a comprehensive array of benefits including dental, health
and life insurance, 4 weeks of vacation annually, and a 401k program.

An Affirmative Action, EEO employer promoting diversity in the workplace.

Apply on-line at:
http://www.schafercorp.com/Careers/open.htm
http://jobs-schafer.icims.com/schafer_jobs/jobs/candidate/job.jsp?jobid=1102
&mode=view


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9, 19 -- Subject: Thermal Ionization Mass Spectrometry Senior Engineer / Scientist Job Opening at Schafer Corporation
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From: christopher.hayden-at-novartis.com
Date: Thu, 19 Jan 2006 08:36:12 -0600
Subject: [Microscopy] RE: Image analysis

Contents Retrieved from Microscopy Listserver Archives
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There are two systems I know of that he may have overlooked:

One is the (dot)Scan, by Soft Imaging Systems
The second is the Mirax system, by Carl Zeiss

I know they can physcially do what you require (overnight, automated
loading and unloading), but I don't know their abilities in terms of
stains.

Hope that helps a little bit!
-Chris


---------------------------------
Christopher Hayden
SPA
Novartis Pharmaceuticals

Hi

Can anyone out there help a colleague?

"I am discussing proposals to install an imaging=20
system here. It is one that can capture both FISH=20
and brown/ H&E stains and then allow an operator,=20
using selected software, to analyse them. The=20
favoured system at present is the Applied Imaging=20
one as it is now undergoing a major facelift.

I am just wondering if there are any other=20
systems on the market right now that can load=20
up 50 slides automatically overnight, scan &=20
store so that an operator can then analyse the=20
images captured the following morning. None of=20
the other major suppliers here in the UK, such as=20
ChromaVision, Meta-systems & Aperio can offer=20
both overnight loading and scanning of both FISH=20
and brown/ blue & pink stains at present."


Med v=E4nliga h=E4lsningar/With best regards

Gareth

http://www.ki.se/biomedlab
e-mail Gareth.Morgan-at-ki.se


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From: bucana-at-audumla.mdacc.tmc.edu
Date: Thu, 19 Jan 2006 18:34:39 -0600
Subject: [Microscopy] viaWWW: Separation of resin from glass tissue culture chamber

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

Dear Yossi,
I have not seen a reply to your inquiry, so I will tell you what I know,
which isn't much. Of all of the mechanisms for contrast in the
back-scattered-electron (BSE) detector, the magnetic domain is the lowest.
First is atomic number, second is channeling contrast and third is magnetic
domain. This means you must eliminate all others to see the magnetic domain.
The sample must be single phase, preferably single crystal and flat,
parallel to the BSE detector, well-polished and, perhaps, electropolished. I
have not done this, only seen talks about it. It takes a lot of beam
current, an optimum working distance for the BSE (about 20 to 15 mm), all
quadrants on and contrast on the BSE amplifier as high as practical. And a
lot of patience.
Good luck.
Regards,
Mary Mager
Electron Microscopist
Department of Materials Engineering
University of British Columbia
6350 Stores Road
Vancouver, B.C. V6T 1Z4
CANADA
Tel: 604-822-5648
Fax: 604-822-3619
e-mail: mager-at-interchange.ubc.ca
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Email: bucana-at-audumla.mdacc.tmc.edu
Name: Corazon D. Bucana

Organization: UT MD Anderson Cancer Center

Title-Subject: [Filtered] Separation of resin from glass tissue culture chamber slides

Question: We have emdedded tissue culture cells grown on tissue culture glass chambered slides using Polybed Epon 812, polymerized at 60C for 2 days. We tried to separate the glass by using liquid nitrogen followed by warm water bath but the glass did not separate from the polymerized resin. Can anyone suggest an alternative procedure to remove the glass from the block? Thanks.

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

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From: M_Jarnik-at-fccc.edu
Date: Thu, 19 Jan 2006 19:23:55 -0600
Subject: [Microscopy] RE: Separation of resin from glass tissue culture chamber slides

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

We grow the cells on a coverslip (round coverslips are better and they
will fit into the chamber) and remove them by hydrofluoric acid
afterwards. Works very well, although the HF is not very pleasant to
work with.

Good luck,

M.

--
Michael Jarnik, Ph.D.
Electron Microscope Facility
Fox Chase Cancer Center
7701 Burholme Ave.
Philadelphia, PA 19111
Tel. 215-728-5675
Fax 215-728-2412




Name: Corazon D. Bucana

Organization: UT MD Anderson Cancer Center

Title-Subject: Separation of resin from glass tissue culture chamber slides

Question: We have emdedded tissue culture cells grown on tissue culture glass chambered slides using Polybed Epon 812, polymerized at 60C for 2 days. We tried to separate the glass by using liquid nitrogen followed by warm water bath but the glass did not separate from the polymerized resin. Can anyone suggest an alternative procedure to remove the glass from the block? Thanks.



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From: redhair-at-stanford.edu
Date: Thu, 19 Jan 2006 22:05:32 -0600
Subject: [Microscopy] RE: Separation of resin from glass tissue culture chamber

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Hi. We too use concentrated hydrofluoric acid to dissolve glass
coverslips. If you use plastic tripour beakers, plastic forceps, and
plenty of rinse water, it doesn't cause problems. Since slides are
thicker than coverslips, they may need may need longer time in the
acid. Coverslips usually dissolve in 20 mins if all the edges are
free of resin. To help the acid get at the free edges, file down the
edges of the embedded slides using a metal file. Then immerse in acid
under the fume hood in a plastic beaker. Check periodically by
dipping in water and looking at the surface( wear nitrile gloves and
work under the hood) The surface should be smooth and shiny with no
patches of undissolved glass( looks like an iceberg melting). Rinse
well under running water and dry in the embedding oven. Good luck!

JoAnn Buchanan
Stanford University school of Medicine
Stanford, CA 94305



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From: dyel-at-mail.nih.gov
Date: Fri, 20 Jan 2006 07:18:43 -0600
Subject: [Microscopy] viaWWW: Immunocytochemistry for TEM

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Email: dyel-at-mail.nih.gov
Name: Louis Dye

Organization: NIH

Title-Subject: [Filtered] Immunocytochemistry for TEM

Question: Greetings ListServers,

Does anyone have a general protocol for labelling anti-lucifer yellow using protein-A gold as a bridging antibody in the Locust (Orthopteran) brain? Should I try to immunogold pre-embedding, using a detergent such as Saponin? Should I dissect out the brain before fixation? I am hoping to use the immunogold labelling to help me visualize neuronal connections. Any advice is greatly appreciated.

Regards,

Louis Dye

Microscopist
NIH, NICHD
Bethesda, MD.

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From: BeverlyM-at-sirna.com
Date: Fri, 20 Jan 2006 07:32:28 -0600
Subject: [Microscopy] viaWWW: liposomes analyzed by Freeze fracture SEM

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Email: BeverlyM-at-sirna.com
Name: Michael Beverly

Organization: Sirna Therapeutics, Inc.

Title-Subject: [Filtered] liposomes analyzed by Freeze fracture SEM

Question: Hello,

I work for a company that is interested in getting some liposomes analyzed by Freeze fracture SEM or another SEM type technique. If there is any way you could put me into contact with some of your members who do this type of work or to post my request on a list serve, etc. it would be extremely helpful.

Thank you,

Michael Beverly
Sirna Therapeutics, Inc.
303-546-8190

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From: nanobio-at-france-optique.org
Date: Fri, 20 Jan 2006 07:55:08 -0600
Subject: [Microscopy] viaWWW: NANOBIO summer school in Cargese France

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Email: nanobio-at-france-optique.org
Name: SociČtČ FranĮaise d'Optique

Organization: Societe Franaise d'Optique

Title-Subject: [Filtered] NANOBIO summer school in Cargese France

Question: Please find enclosed a flyer on the NANOBIO summer school in Cargese,
Corsica, France from July 17th till 29th, 2006.

Watching single biological molecules at work is now possible and the
unravelled features of the molecular machinery at the nanoscale are
currently changing our view of the cell. Though, bridging the gap
between nano- and micro-world is not easy: how to relate single molecule
events to biological functions at the cell level? how do molecular
structures assemble, coordinate and integrate in the wholeÖ? Thanks to
physicists and biologists, we will address these questions and try to
give some answers. Speakers will present recent concepts and methods in
physics and biology dedicated to single molecule observation and
description of the cell at the sub-micron level. Particular emphasis
will be given to the optical methods, which allow the observation of
live cells in physiological conditions.

This school is part of the Nanosciencestech Marie Curie Series of Events:
http://www.france-optique.org/Nanosciencestech.html

The students, including PhDs, post-docs, and young researchers, will be
encouraged to present their own work (also in adjacent fields) during
poster sessions.

A European and more generally an international audience is targeted, and
all lectures in the summer school will be given in English.

Important dates:
Pre-registration: Application deadline 28 february 2006
Selected participant will be informed before the end of march

Registration and paying attendees deadline: 30 April 2006

All correspondence should be sent to

SociČtČ FranĮaise d'Optique
Nanobio summer school
c/o FranĮoise CHAVEL
Centre Scientifique d'Orsay - B’t. 503
91403 ORSAY cedex France

Phone : ++33 1 69 35 88 33
Fax : ++33 1 69 85 35 65 or ++33 1 69 35 87 33
mail : nanobio-at-france-optique.org

Sponsors :

EUROPEAN COMMUNITY
SOCIETE FRANCAISE DķOPTIQUE
GDR 2588 ń Microscopie Fonctionnelle
CARL ZEISS FRANCE


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From: phillipst-at-missouri.edu
Date: Fri, 20 Jan 2006 08:48:26 -0600
Subject: [Microscopy] Re: viaWWW: Separation of resin from glass tissue

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next time, you should only polymerize for about 7-8 hours before popping
the cells off. I then usually re-embed the pieces that come off and
polymerize using standard times.


At 06:43 PM 01/19/06, you wrote:



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From: lenoska1-at-seznam.cz
Date: Fri, 20 Jan 2006 09:10:24 -0600
Subject: [Microscopy] =?us-ascii?Q?fluorescence=20microscopy?=

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Name: Zuzana Lenochova

Organization: PrF UK

Title-Subject: positive TUNEL in negative control

Question:

Greetings ListServers,
does anybody have any experience using TUNEL assay on plants? I use the kit from Roche, nucleotides dyed with TMR, and have a problem that all the seccions (free-hand,fixed,penetrated with Triton and pectinase/cellulase solution), even those treated just with labeling solution without enzymes, are TUNEL positive. Do you know any reasonable explanation how could the nucleotides bind on the nucleus (and slightly on plasmatic membrane) without enzymes added?

Best regards,

Zuzana Lenochova
Charles University
Prague

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From: larry-at-cymru.freewire.co.uk
Date: Fri, 20 Jan 2006 14:31:08 -0600
Subject: [Microscopy] Re: Magnetic contrast with 4QBSED

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} } Hi,
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} } I am interested to study magnetic structure (type II magnetic contrast) by
} } SEM. Our SEM equipped with a Four Quadrant Backscattered Electron
} Detector. I
} } will appreciate if you have experience using this type of detector for
} } magnetic domain structure to guide us with the recommended working
} parameters
} } as combinations of quadrant setting, working distance, tilt and etc.
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} }
} } Yossi.
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I've never done this and I'm trying to remember an illustration from
an old JEOL SEM brochure, which described an attachment for magnetic
domain imaging.

From what I recall, the experimental setup was similar to that used
now for EBSD. Sample at a high tilt with a detector to one side. In
this case, a BSE detector. The idea was that the weak effect of the
magnetic domains caused a small divergence of BS electrons. By
positioning the detector a long way from the sample and at a high
angle from the incident beam, the divergence was amplified.
--
Larry Stoter

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From: GBurgess-at-exchange.hsc.mb.ca
Date: Fri, 20 Jan 2006 16:10:03 -0600
Subject: [Microscopy] Wrinkled LM Sections

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Even though I am very experienced in Electron Microscopy, there are times
when I cut LM sections that are extremely wrinkled, and they look like
cracked glass. I normally don't experience this, but when it happens, even
considering every possible variable, I'm still not exactly sure what might
be at work here to give this sort of result.

I'm looking for ideas here as to what might be causing this effect. What do
people think?


Garry Burgess
Charge Technologist - Electron Microscopy
Health Sciences Centre
Winnipeg, Canada.

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From: W.Muss-at-salk.at
Date: Sat, 21 Jan 2006 04:43:59 -0600
Subject: [Microscopy] Re: Wrinkled LM Sections

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Good morning, all,

Dear Garry,

in my experience (I am familiar with large semithin sections = up to 5 x 5
mm from human diagnostic samples for some 20 years now) your question
cannot be answered with one sentence.
There are several parameters to be included in a thoroughly checking of
causes, starting from the type of tissue, via fixation, (full) dehydration,
intermedium (full evaporation/complete exchange by resin), type, quality
[polymerisation, hardness] of resin, quality of knife edge, sectioning
parameters (knife, cutting angle, speed) and type of transfer of sections
from the water trough to the slide, and last but not least, "special
treatment" of the floating section on a water drop (i.e., for example,
trying to spread sections by xylene vapor [other - more healthier -
vapors?] or gentle - longer - warming up by means of a light bulb
positioned above the section/slide).
So IMO, you have to discriminate the problem(s) from the whole processing
schedule.......(;-(.....

You tell us that you face problems not everytime, but sometimes.....
Could you give us some examples (tissue type, some hints on chemicals
/resin you use?) where you get such results? (not to forget the dimensions
of the tissue blocks and type of knife used)....

Perhaps there is a simple solution......who knows,


best regards and have a nice weekend,

Wolfgang Muss

Salzburg, Austria

Paracelsus Medical Private University (PMU)
Institute of Pathology
Electron Microscopy Lab
Muellner Hauptstrasse 48
A-5020 SALZBURG, Austria/Europe
Phone work: +43+662+4482+4720
Mobile phone work:+43+662+4482-57704
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Von: GBurgess-at-exchange.hsc.mb.ca[SMTP:GBurgess-at-exchange.hsc.mb.ca]
Antwort an: GBurgess-at-exchange.hsc.mb.ca
Gesendet: Freitag, 20. Janner 2006 23:52
An: W.Muss-at-salk.at
Betreff: [Microscopy] Wrinkled LM Sections

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I'm looking for ideas here as to what might be causing this effect. What
do
people think?


Garry Burgess
Charge Technologist - Electron Microscopy
Health Sciences Centre
Winnipeg, Canada.

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From: wong-at-msg.ucsf.edu
Date: Sat, 21 Jan 2006 08:02:30 -0600
Subject: [Microscopy] viaWWW: Making formvar thicker

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Email: wong-at-msg.ucsf.edu
Name: Mei Lie Wong

Title-Subject: [Filtered] formvar

Question: When using formvar in ethylene dichloride, is the any way to make the films slightly thicker. I know when using formvar in chloroform, you can just drain slower or faster to make films thinner or thicker. Is there something like this for the ethylene dichloride formula? I have tried several different times but so far haven't hit on anything totally satisfactory.

Thanks in advance.

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From: cgarber-at-2spi.com
Date: Sat, 21 Jan 2006 10:41:37 -0600
Subject: [Microscopy] Formvar filmed grids: controlling thickness

Contents Retrieved from Microscopy Listserver Archives
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Mei Lie Wong wrote:
========================================================
Question: When using formvar in ethylene dichloride, is the any way to make
the films slightly thicker. I know when using formvar in chloroform, you
can just drain slower or faster to make films thinner or thicker. Is there
something like this for the ethylene dichloride formula? I have tried
several different times but so far haven't hit on anything totally
satisfactory.
=======================================================
This is discussed on
http://www.2spi.com/catalog/submat/sup_film6.html

Disclaimer: SPI Supplies manufactures Formvar coated grids for customers
and we disclose some of the tricks we use for making thicker or thinner
films when using ethylene dichloride.

Chuck

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

Charles A. Garber, Ph. D. Ph: 1-610-436-5400
President
SPI SUPPLIES FAX: 1-610-436-5755
PO BOX 656 e-mail:cgarber-at-2spi.com
West Chester, PA 19381-0656 USA
Cust.Service: spi2spi-at-2spi.com

Look for us!
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From: bigelow-at-engin.umich.edu
Date: Sat, 21 Jan 2006 11:29:44 -0600
Subject: [Microscopy] RE: Starting Ion Pumps

Contents Retrieved from Microscopy Listserver Archives
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I was interested in the comments about banging on the sides of a
sputter ion pump with a hammer or screw driver to overcome the
startup problem that arises when a whisker or flake of titanium has
formed and shorted out the path between the cathode and anode of the
pump. Another way to treat this problem is discussed on p. 295 of my
book 'Vacuum Methods in Electron Microscopy' (available from SPI,
Ladd, M.E. Taylor, etc. For a description see:
www.2spi.com/catalog/books/book48.html). This involves allowing the
pressure in the pump to rise slightly above 1 Pa (10-2 Torr) and then
turning on the high voltage power for a few seconds for three or four
times. Be careful not to do this long enough to damage the power
supply.
--
Wilbur C. Bigelow, Professor Emeritus
Materials Sci. & Engr., Univ. of Michigan
Ann Arbor, Michigan 48109-2136
e-mail: bigelow-at-engin.umich.edu;
Fx:734-763-4788; Ph:734-662-5237
Address mail to: 1136 Mixtwood Rd.
Ann Arbor, MI 48103-3035

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From: gary-at-gaugler.com
Date: Sat, 21 Jan 2006 12:09:23 -0600
Subject: [Microscopy] Re: RE: Starting Ion Pumps

Contents Retrieved from Microscopy Listserver Archives
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Yes, that might also work. However, if I just got
through with an 8 hour bakeout, why would I want to
degrade vacuum in the gun chamber? I would either
never get the chamber pumped down or it would take a
very long time to reach terminal vacuum.

The idea is to get vacuum as good as possible and then
isolate the gun chamber before turning on the pump. Even
so, startup current would be high.

I've used the hair dryer most of the time. Then, sometimes
whack the pump with the screwdriver handle. This is only for
the small pumps. For some reason, they don't start as readily
as the larger ones.

gary g.


At 09:48 AM 1/21/2006, you wrote:



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From: murphyjudy-at-comcast.net
Date: Sun, 22 Jan 2006 17:23:22 -0600
Subject: [Microscopy] We Remember Del Philpott

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Celebration of life for
Delbert E. Philpott
September 24, 1923 - December 11, 2005

Yesterday at the Liberty Lodge Masonic Center in Santa Clara, CA, we
honored the life and friendship of Dr. Del Philpott, a lifelong
microscopist and soldier who truly believed in the perpetuation of peace
and worked for that goal his entire life.

His wife, Donna, agreed to our reprinting of a few of his many
achievements which she listed in the memorial pamphlet.

Del was born and raised in Omro, Wisconsin. Dr. Philpott was a World
War II Veteran who served with the Fighting 69th Infantry Division as
they raced across Europe to link up with the Russian 58th Guards at the
Elbe River in Germany in April of 1945. The famous link-up photo, by
photographer Allan Jackson of 3 Americans (including Del), and 3
Russians shaking hands was printed in newspapers across the United
States. The soldiers however didn't see the photo because the war
hadn't yet ended. Fellow Infantryman Sam Popkins, who had helped Allan
recruit soldiers for the photo, later told Del about it and confirmed
that Del was one the soldiers in the photo. Del was awarded the Bronze
Star and Purple Heart Medals for his service.

After the war, Del completed his Bachelor's Degree in chemistry at
Indiana University in 1948 and his Master's Degree in 1949. As a
Research Associate at the University of Illinois Medical School in
Chicago from 1949 to 1952, he established the first electron microscope
facility for the medical school. He was Head of Electron Microscopy,
Marine Biological Lab and Institute for Muscle Research, Woods Hole, MA
from 1952 to 1963, where he established the electron microscope facility
for the 2 Institutes.

A pioneer in the field of Electron Microscopy, he directed and ran
research projects under Nobel Laureate Dr. Albert Szent-Gyorgyi in the
winter and for the Marine Biological Lab at Woods Hole during the
summer. He built his own ultramicrotome for ultra-thin sectioning.

He published the first paper on ultrastructure of the human heart with
Dr. Bruno Kish and published the first electron micrographs of a protein
crystal isolated from muscle. He demonstrated that ribosomes travel
from the nucleus to the cytoplasm via the nuclear pores. Del got his
PhD in cytology in 1963 at Boston University.

While Professor of Biochemistry at the University of Colorado from 1963
to 1965, he established and ran the electron microscope laboratory at
the medical school. In 1966, he was Head of the Department of Electron
Microscopy and Co-Director of the Institute for Biomedical Research at
Mercy Hospital in Denver, CO.

Del came to California in the 60's to work as a Research Scientist as
the Head of the Ultrastructure Laboratory at the Moffett Field NASA Ames
Research Center. He inspected lunar soil for signs of life and had
experiments flown on both American and Russian spacecraft including
Apollo 17, Cosmos 736, Cosmos 936, and SL-3. He developed a fixative
for space flight that preserves the ultrastructure of tissues for over 4
years without the need for embedding. Del retired from NASA in 1990,
but worked under contract with the Lockheed Martin Company at NASA Ames
as a Research Associate whose duties included assisting visiting Russian
scientists with space related experiments.

During his professional career, Del authored over 230 scientific papers
and wrote articles for several books and magazines.

In 1995, Del and his wife Donna co-edited the book "Hands Across The
Elbe", stories by American and Russian veterans about the link up that
cut Fascist Germany in half in April 1945.

Del was one of 10 veterans chosen to be guests at the Russian
government's May 9, 2005, 60th Anniversary Celebration of the end of
World War II. He was selected to be seated at the table with the
President of China, President Putin, and President and Mrs. Bush at the
Kremlin Reception following the Parade e in Red Square.

Del's interests included flying, raising dogs, ham radio, ceramics,
spinning, photography, woodcarving, magic, and travel. He held offices
in many organizations including several scientific societies, veterans'
organizations, and The Unique Boutique at the Sunnyvale Senior Center.
He was a member of Liberty Lodge $299, Valley Star Chapter #250 O.E.S.,
and the San Jose Scottish Rite.

He wanted to be remembered for his part in developing interaction
between the American and Russian scientists in space research. He
believed that interactions among the citizens of all nations are
necessary for the perpetuation of peace and felt that the "Spirit of the
Elbe" encourages this vision.

Thank you Donna for putting together such an information-packed
bibliography.

I knew Del from the late 60's from the microscopy society. He was a
wonderful friend and had an uncanny sense of humor. I would like to
share 2 of Del's favorite stories. And for those of you who knew Del,
you know he had LOTS of stories.

I can no longer remember the exact year, but it was in the 70's.
Someone sent in a bogus abstract for the EMSA meeting with a crazy title
and crazy names of authors. Unfortunately I no longer remember those
either. However the following year, as you can imagine, EMSA was trying
to review the abstracts or at least look more carefully at the titles
and authors. That year Del sent in an abstract about research on lunar
samples. Well, whoever reviewed his abstract decided that no one could
possibly be named Delbert Philpott and certainly no one was looking at
moon samples, SO, his paper was rejected!!

Del's sense of humor was ever present. Before he knew Donna, again in
the 70's sometime, Del was at the EMSA meeting telling us about a
Valentine's present he got for his girlfriend. He bought some dog
biscuits (yes dog biscuits), had them chocolate coated, put them in
those fancy white wrappers, and put them in a standard Valentine's Candy
Box. He gave them to his girlfriend. The next year I saw Del, I asked
him how his girlfriend liked her Valentine's gift. He said "Well, I
actually haven't seen her since then. I guess she didn't like that
brand dog biscuits"!!!!

Del is gone, but his loving sense of humor, and desire for the promotion
of peace through interactions among citizens of all nations will always
be with us.
We love you Del, and will miss you always.

God Bless Us All,
Judy Murphy


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From: nairvinods-at-gmail.com
Date: Sun, 22 Jan 2006 17:45:06 -0600
Subject: [Microscopy] Wrinkled LM Sections

Contents Retrieved from Microscopy Listserver Archives
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Dear Gary,
I have been working with LR white and have been facing similar
problems. They are more so only when I immuno label those sections.
Then again my sections are more than 2 mm in size and they are thin
sections. I have managed to reduce the wrinkles by changing the
washing techniques between immunolabelling steps but that hasnt
completely eliminated the wrinkles. I have observed that LR white
section around 1mm wide didnt have so many wrinkles.
Unfortunately for me I cant have thicker sections or smaller sections
to avoid this problem. I cannot use a different resin since it
interfers with my immunolabeling. SO the bottom line being, I have
learnt to live with it unfortunately.
But I have a feeling that smaller sections do better and the wrinsing
the grids in a drop of water/ buffer on a petridish does help reduce
the wrinkles greatly if not eliminate it.

Any advice on photoshop micracles of ironing out the wrinkles would be
appreciated. I hope facelifting TEM pics will not be frowned upon by
the microscopy community.

Looking forward for comments and suggestion.

Regards,
Vinod Nair
Graduate Student
Dept. of Biology
New Mexico State University

On 1/21/06, W.Muss-at-salk.at {W.Muss-at-salk.at} wrote:
}
}
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} Good morning, all,
}
} Dear Garry,
}
} in my experience (I am familiar with large semithin sections = up to 5 x 5
} mm from human diagnostic samples for some 20 years now) your question
} cannot be answered with one sentence.
} There are several parameters to be included in a thoroughly checking of
} causes, starting from the type of tissue, via fixation, (full) dehydration,
} intermedium (full evaporation/complete exchange by resin), type, quality
} [polymerisation, hardness] of resin, quality of knife edge, sectioning
} parameters (knife, cutting angle, speed) and type of transfer of sections
} from the water trough to the slide, and last but not least, "special
} treatment" of the floating section on a water drop (i.e., for example,
} trying to spread sections by xylene vapor [other - more healthier -
} vapors?] or gentle - longer - warming up by means of a light bulb
} positioned above the section/slide).
} So IMO, you have to discriminate the problem(s) from the whole processing
} schedule.......(;-(.....
}
} You tell us that you face problems not everytime, but sometimes.....
} Could you give us some examples (tissue type, some hints on chemicals
} /resin you use?) where you get such results? (not to forget the dimensions
} of the tissue blocks and type of knife used)....
}
} Perhaps there is a simple solution......who knows,
}
}
} best regards and have a nice weekend,
}
} Wolfgang Muss
}
} Salzburg, Austria
}
} Paracelsus Medical Private University (PMU)
} Institute of Pathology
} Electron Microscopy Lab
} Muellner Hauptstrasse 48
} A-5020 SALZBURG, Austria/Europe
} Phone work: +43+662+4482+4720
} Mobile phone work:+43+662+4482-57704
} Fax-No. at work: ++43+662+4482-882 ext (please, only by indicating: "c/o
} W.Muss")
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}
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} ------------------------------------------------------------------------
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} } http://www.scur.org {
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}
}
}
}
} ----------
} Von: GBurgess-at-exchange.hsc.mb.ca[SMTP:GBurgess-at-exchange.hsc.mb.ca]
} Antwort an: GBurgess-at-exchange.hsc.mb.ca
} Gesendet: Freitag, 20. Janner 2006 23:52
} An: W.Muss-at-salk.at
} Betreff: [Microscopy] Wrinkled LM Sections
}
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} Even though I am very experienced in Electron Microscopy, there are times
} when I cut LM sections that are extremely wrinkled, and they look like
} cracked glass. I normally don't experience this, but when it happens, even
} considering every possible variable, I'm still not exactly sure what might
} be at work here to give this sort of result.
}
} I'm looking for ideas here as to what might be causing this effect. What
} do
} people think?
}
}
} Garry Burgess
} Charge Technologist - Electron Microscopy
} Health Sciences Centre
} Winnipeg, Canada.
}
} This e-mail and/or any documents in this transmission is intended for the
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From: murphyjudy-at-comcast.net
Date: Sun, 22 Jan 2006 22:53:44 -0600
Subject: [Microscopy] Re: viaWWW: Making formvar thicker

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

Mei Lie,
Making formvar films in ethylene dichloride
We used these films for 30 yrs and controlled the thickness mostly by
the percentage of formvar to ethylene dichloride. What percentage are
you using?

Judy



Judy Murphy, PhD
Microscopy Training, Imaging, and Lab Design
Stockton, CA 95219
murphyjudy-at-comcast.net





wong-at-msg.ucsf.edu wrote:

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From: murphyjudy-at-comcast.net
Date: Sun, 22 Jan 2006 23:08:27 -0600
Subject: [Microscopy] Re: We Remember Del Philpott

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

A web site was brought to my attention that is a collection of
photographs by him (and several of him) during Del's time with Albert
Szent-Gyorgyi. I found it very interesting and rich in the history of
science.

http://profiles.nlm.nih.gov/WG/Views/Exhibit/other/visuals.html

Enjoy,
Judy Murphy

Thank you Peter for sharing the information AND hold on to that signed
copy of Crossing the Elbe!


murphyjudy-at-comcast.net wrote:

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From: pjordan-at-dslextreme.com
Date: Sun, 22 Jan 2006 23:53:28 -0600
Subject: [Microscopy] Iongetter pump for Zeiss 109

Contents Retrieved from Microscopy Listserver Archives
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Hi:
I am restoring a Zeiss 109 TEM where the Leybold IZ-80 iongetter pump has
been removed, however they left the magnets. The pump does not have to be in
working condition. If you have on lying around and are willing to part with
it or sell it please let me know.
Peter Jordan/EMSI


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From: nairvinods-at-gmail.com
Date: Mon, 23 Jan 2006 00:03:23 -0600
Subject: [Microscopy] Re: Wrinkled LM Sections

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Dear Gary,
Let me begin with apologising for not reading the email correctly. You
can see what I have been pondering over from my reply with regards to
LR white resin:( Guess it was desperation that made me misread LM as
LR and link it to the wrinkeling problems I have been facing.
Having said that I feel very foolish now for being hasty and replying
to your email query. My sincere apologies.

But then again I am open for suggestions,comments or insight into the
problem I am facing.
regards,
Vinod


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From: tivol-at-caltech.edu
Date: Mon, 23 Jan 2006 11:37:41 -0600
Subject: [Microscopy] Re: viaWWW: Making formvar thicker

Contents Retrieved from Microscopy Listserver Archives
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On Jan 21, 2006, at 6:03 AM, wong-at-msg.ucsf.edu wrote:

} Question: When using formvar in ethylene dichloride, is the any way to
} make the films slightly thicker. I know when using formvar in
} chloroform, you can just drain slower or faster to make films thinner
} or thicker. Is there something like this for the ethylene dichloride
} formula? I have tried several different times but so far haven't hit
} on anything totally satisfactory.
}
Dear Mei Lie,
Using a more concentrated formvar solution should do it, and if you
only have the pre-disolved formvar, you could try either letting some
of the solvent evaporate or double dipping. For finer variations you
might try letting the formvar drain at a shallower angle, which will
drain it slightly slower. Good luck.
Yours,
Bill Tivol, PhD
EM Scientist and Manager
Cryo-Electron 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: glenmac-at-u.washington.edu
Date: Mon, 23 Jan 2006 11:39:42 -0600
Subject: [Microscopy] fiber illuminator for Opni-1

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

Dear All,
We have an older Zeiss Opni-1 surgical microscope and we'd like to
replace its broken tungsten illuminator with a fiber optic
illuminator. this would preserve the large illuminated field
diameter and be adjustable in angle. the usual web searches have
shown that retrofits were made for this scope, but haven't provided
any useful information on performance or options. It is already
used in conjunction with bifurcated light guides, but we have users
who need the stock style of illumination.

Thanks,
Glen


Glen MacDonald
Core for Communication Research
Virginia Merrill Bloedel Hearing Research Center
Box 357923
University of Washington
Seattle, WA 98195-7923 USA
(206) 616-4156
glenmac-at-u.washington.edu

************************************************************************
******
The box said "Requires Windows 95 or better", so I bought a Macintosh.
************************************************************************
******



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From: vfink-at-shaw.ca
Date: Mon, 23 Jan 2006 12:04:18 -0600
Subject: [Microscopy] thermal conductivity of thin film measurement techniques

Contents Retrieved from Microscopy Listserver Archives
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Dear All:

I need to measure thermal conductivity of thin, 50-200 nm film. I would
appreciate any suggestions regarding to equipment, techniques, and
literature references.

Thank you,

Victoria Fink
vfink-at-shaw.ca



==============================Original Headers==============================
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From: vfink-at-shaw.ca
Date: Mon, 23 Jan 2006 12:04:25 -0600
Subject: [Microscopy] thermal conductivity of thin film measurement techniques

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


Dear All:

Inked to measure thermal conductivity of thin, 50-200 nm film. I would
appreciate any suggestions regarding to equipment, techniques, and
literature references.
Thank you,

Victoria Fink
vfink-at-shaw.ca



==============================Original Headers==============================
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From: sales-at-laddresearch.com
Date: Mon, 23 Jan 2006 12:39:16 -0600
Subject: [Microscopy] Re: viaWWW: Making formvar thicker

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

A number of our customers for coated grids request a variety of
thicknesses. What we do for thicker coated grids is increase the
percentage of formvar in ethylene dichloride.

Mike Bouchard

Disclaimer: Ladd Research sells Formvar coated grids and the
essentials to make your own.


Ladd Research
83 Holly Court
Williston, VT 05945

On-line Catalog: http://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 09:46 AM 1/21/2006, wong-at-msg.ucsf.edu wrote:



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

Ladd Research
83 Holly Court
Williston, VT 05945

On-line Catalog: http://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



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From: underwoo-at-u.washington.edu
Date: Mon, 23 Jan 2006 13:08:15 -0600
Subject: [Microscopy] Cryoprotection causing shrinkage artifact

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

To Fellow Microscopists,

I am curious about the possible skinkage artifact that may or may not occur
when cryoprotecting lightly fixed (4% para / 0.1% glut) watery tissues
(embryonic skin) in 20% PVP / 1.7M sucrose for ultrathin cryomicrotomy? Does
the cryoprotectant replace the water in a controlled way and keep the tissue
fat and happy or does it draw the water out and collapse the tissue? I would
love any information on this subject or a possible reference.

Robert Underwood
University of Washington
Dermatology


==============================Original Headers==============================
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From: jae5-at-lehigh.edu
Date: Mon, 23 Jan 2006 13:25:49 -0600
Subject: [Microscopy] Post doctoral position at Lehigh

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

Postdoctoral Position,
Department of Materials Science and Engineering
Lehigh University,

Available March 2006. Opening for post-doc to work on project involving
processing of patterned sapphire substrates via oxidation and solid
state conversion of a metallic Al coating. Person would be responsible
for processing and EBSD/TEM characterization of both the substrates,
together with TEM of MOCVD grown GaN layers to determine the defect
density. Expertise in materials processing and electron microscopy
required, experience with thin film processing a plus. Please forward
resume by e-mail to Prof. Helen Chan (Helen.Chan-at-lehigh.edu) and / or
Prof. Richard Vinci (rpv2-at-lehigh.edu).

...........

Department of Materials Science and Engineering
Lehigh University
5 East Packer Avenue
Bethlehem
Pennsylvania 18015-3195


==============================Original Headers==============================
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From: hagglundk1-at-nku.edu
Date: Tue, 24 Jan 2006 09:02:20 -0600
Subject: [Microscopy] SEM x-ray peak ratios

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

I have a colleague who recently experienced some down time on his SEM.
The system went through repeated cycles of pump down, and venting and
was eventually left powered down while waiting on a computer
replacement.

After getting everything operating again, an oil film was found that had
built up on the x-ray detector thin window. This was initially causing
an unacceptable amount of background in the 0-3kV range of the x-ray
signal as well as an overall low count rate. The collimator was removed
and cleaned, which corrected the noise and count rate. They are in the
process of replacing the roughing lines and cleaning the chamber to
prevent further contamination. Now, when calibrating with a copper
standard, the ratio of the L line signal versus the K lines is
dramatically favoring the L. This was not the case before.

Any ideas why this might be happening and how to correct it?

Karl Hagglund
Biological Sciences, SC-102B
Northern Kentucky University, Nunn Drive
Highland Heights, KY 41099
859-572-5238
http://semlab.nku.edu


==============================Original Headers==============================
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From: maloneyb-at-fiu.edu
Date: Tue, 24 Jan 2006 13:40:31 -0600
Subject: [Microscopy] Phillips 300 TEM calibration

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

Does anyone remember a quick and easy way to calibrate - it has what 25
taps - so if anyone remembers how to do the calculations, please let me
know ASAP.
Thanks
Barbara

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From: edelmare-at-muohio.edu
Date: Tue, 24 Jan 2006 14:11:14 -0600
Subject: [Microscopy] Cryo-ultramicrotomy workshop?

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

I have had a request from a user looking for a cryo-
ultramicrotomy workshop. Is anyone offering one in the next year or
know of one? Thanks!



Richard E. Edelmann, Ph.D.
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

"WE ARE MICROSOFT.
RESISTANCE IS FUTILE.
YOU WILL BE ASSIMILATED."

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==============================End of - Headers==============================




From: emlabservices-at-cox.net
Date: Tue, 24 Jan 2006 16:34:06 -0600
Subject: [Microscopy] Emispec Systems/FEI Support?

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

Hi All,

Several years ago, Emispec Systems was purchased by FEI Company of
Hillsboro, OR. Recently I've been contacted by several customers who in the
past had purchased these data acquisition systems for their EM's and stated
they are having difficulty making contact with the responsible person or
department at FEI for upgrades and maintenance support.

I'm wondering if parties who are present owners of Emispec ES Vision systems
have had success in seeking this support and can provide contact information
or advise of appropriate channels to go through to obtain system support.

Please respond off list if you can shed some light on this.

Regards,

Bob Roberts
EM Lab Services, Inc.
449 NW 62nd St.
Topeka, Kansas 66617-1780
785.246.1232 voice
785.246.0168 fax
www.emlabservices.com



==============================Original Headers==============================
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From: fmalherbe-at-swin.edu.au
Date: Wed, 25 Jan 2006 07:33:17 -0600
Subject: [Microscopy] [Filtered] AskAMicroscopist: Printer for an SEM

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

Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (fmalherbe-at-swin.edu.au) from http://www.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Tuesday, January 24, 2006 at 18:23:45
---------------------------------------------------------------------------

Email: fmalherbe-at-swin.edu.au
Name: Francois Malherbe

Organization: Swinburne University of Technology

Education: Graduate College

Location: Melbourne, Victoria, Australia

Question: We are purchasing a new SEM, what are the basic specs you would recommend for the printer:

- inkjet, laser or other?
- USB, serial?
- dpi?

Thank you for your help.

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

==============================Original Headers==============================
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9, 12 -- Subject: [Filtered] AskAMicroscopist: Printer for an SEM
9, 12 -- Content-Type: text/plain; charset="us-ascii"
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From: sallwein-at-ncifcrf.gov
Date: Wed, 25 Jan 2006 07:34:04 -0600
Subject: [Microscopy] [Filtered] MicroscopyListserverviaWWW: Employment Opportunity to

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

This Question/Comment was submitted to the Microscopy Listserver
using the WWW based Form at http://www.microscopy.com/MLFormMail.html
---------------------------------------------------------------------------
Remember this posting is most likely not from a Subscriber, so when replying
please copy both sallwein-at-ncifcrf.gov as well as the MIcroscopy Listserver
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Email: sallwein-at-ncifcrf.gov
Name: Stacy Allwein

Organization: SAIC-Frederick, Inc.

Title-Subject: [Filtered] Employment Opportunity to work in Support of the National Cancer Institute at SAIC-Frederick, Inc.

Question: SAIC-Frederick, Inc., working in support of the National Cancer Institute at Frederick, seeks an experienced Research Associate to be responsible for independent imaging TEM and SEM samples in a core facility, processing samples and coordinating GLP samples in the lab, processing, sectioning and imaging core samples, training, and conduct collaborative projects with the National Cancer Institute and the National Institutes of Health. Position requires a BS degree and at least four years of experience with TEM and/or SEM sample processing and imaging.

Please visit our web site at http://saic.ncifcrf.gov and refer to opportunity KMB134306 to apply on line.

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

==============================Original Headers==============================
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7, 13 -- Subject: [Filtered] MicroscopyListserverviaWWW: Employment Opportunity to
7, 13 -- work in Support of the National Cancer Institute at SAIC-Frederick
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From: murphyjudy-at-comcast.net
Date: Wed, 25 Jan 2006 09:51:52 -0600
Subject: [Microscopy] Re: Cryo-ultramicrotomy workshop?

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

The 5th International Cryo-EM Course is I believe June 6-17, 2006 in
Vancouver, Canada at University of British Columbia by Dr. Elaine
Humphrey from UBC and Dr. Kent McDonald, from UC, Berkeley with
international speakers.
This is an excellent course and caters to the needs of the particular
students. Be sure to check with Elaine about dates.
For more info email
Dr. Elaine Humphrey
ech-at-interchange.ubc.ca

Information from the previous course is listed at
http://www.emlab.ubc.ca/CryoEM2005/index.html

Judy


Judy Murphy, PhD
Microscopy Training, Imaging, and Lab Design
Stockton, CA 95219
murphyjudy-at-comcast.net







edelmare-at-muohio.edu wrote:

} ----------------------------------------------------------------------------
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==============================Original Headers==============================
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From: murphyjudy-at-comcast.net
Date: Wed, 25 Jan 2006 09:57:46 -0600
Subject: [Microscopy] Re: Cryo-ultramicrotomy workshop?

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

Link to current cryocourse June 6-15, 2006 at University of British
Columbia, Vancouver, BC
Here is the link to the current cryo course in 2006.
http://www.emlab.ubc.ca/CryoEM2006/index.html


Judy Murphy wrote:

} The 5th International Cryo-EM Course is I believe June 6-17, 2006 in
} Vancouver, Canada at University of British Columbia by Dr. Elaine
} Humphrey from UBC and Dr. Kent McDonald, from UC, Berkeley with
} international speakers.
} This is an excellent course and caters to the needs of the particular
} students. Be sure to check with Elaine about dates.
} For more info email
} Dr. Elaine Humphrey
} ech-at-interchange.ubc.ca
}
} Information from the previous course is listed at
} http://www.emlab.ubc.ca/CryoEM2005/index.html
}
} Judy
}
}
} Judy Murphy, PhD
} Microscopy Training, Imaging, and Lab Design
} Stockton, CA 95219
} murphyjudy-at-comcast.net
}
}
}
}
}
}
}
} edelmare-at-muohio.edu wrote:
}
} } ----------------------------------------------------------------------------
} }
} } The Microscopy ListServer -- CoSponsor: The Microscopy Society of
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} } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} } ----------------------------------------------------------------------------
} }
} }
} } I have had a request from a user looking for a cryo-
} } ultramicrotomy workshop. Is anyone offering one in the next year or
} } know of one? Thanks!
} }
} }
} }
} } Richard E. Edelmann, Ph.D.
} } 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
} }
} } "WE ARE MICROSOFT.
} } RESISTANCE IS FUTILE.
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==============================Original Headers==============================
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From: rcommon-at-msu.edu
Date: Wed, 25 Jan 2006 10:07:43 -0600
Subject: [Microscopy] Olympus BX41 Manual

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


I am in need of a manual for the Olympus BX41 microscope, particularly the
epifluorescence unit, U-L100HGAPO. If anybody knows of on-line
instructions, or how I could obtain a copy of the manuals, I would be very
grateful. I will be happy to pay any copying and mailing charges, of
course.

Ralph Common
Dept. of Physiology
Michigan State University


==============================Original Headers==============================
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4, 24 -- Subject: Olympus BX41 Manual
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From: shem-at-laurentian.ca
Date: Wed, 25 Jan 2006 14:51:27 -0600
Subject: [Microscopy] Etching of Cu-Ni matte for reflected light microscopy

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

Hello Everyone,
Is there anyone out there who has experience etching Cu and Ni sulfides, in order
to see compositional zoning ?

The phases I am interested in are: Cu2S and Ni3-XS2. In both these compounds Ni and Cu substitute
for each other. It is these substitutions which I suspect change the etching behaviour of the compounds.
Etching could thus be a quick and dirty alternative to excessive X-ray mapping/analysis.

Any input is welcome !

Thanks,

Skage Hem



_______________________________________________________________

Skage Hem
Research Scientist, Ph.D.
CAF, Department of Earth Sciences
Laurentian University
Ramsey Lake Road
Sudbury, ON
Canada
P3E 2C6

ph. 705-675-1151 x4040
cell. 705-562-7321
fax 705-675-4898

http://laurentian.ca/geology/Research/hem.html



==============================Original Headers==============================
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13, 17 -- From: "Skage Hem" {shem-at-laurentian.ca}
13, 17 -- To: {microscopy-at-microscopy.com}
13, 17 -- Subject: Etching of Cu-Ni matte for reflected light microscopy
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From: opmills-at-mtu.edu
Date: Wed, 25 Jan 2006 14:51:54 -0600
Subject: [Microscopy] estimates of value

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

Hi,

I'm trying to establish a value on a functional Link (Oxford) EDS
detector and LEMAS stage automation system. Detector has been
rebuilt 5 years ago.

Before I can sell it I have to get a few opinions of value for my
property management department. Can any of you help me?

Please respond to me directly.

Thanks!

Owen

Owen P. Mills
Director, Materials Characterization & Fabrication Facilities
Electron Optics Engineer, Applied Chemical & Morphological Analysis
Laboratory

Materials Science & Engineering
Michigan Technological University
Rm 512 M&M Bldg.
Houghton, MI 49931
PH 906-369-1875
FAX 906-487-2934
mailto:opmills-at-mtu.edu
http://www.mm.mtu.edu/~opmills



==============================Original Headers==============================
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10, 31 -- To: Microscopy Listserver {Microscopy-at-microscopy.com}
10, 31 -- From: "Owen P. Mills" {opmills-at-mtu.edu}
10, 31 -- Subject: [Microscopy] estimates of value
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From: smalinskas-at-yahoo.com
Date: Wed, 25 Jan 2006 15:39:14 -0600
Subject: [Microscopy] Re: Etching of Cu-Ni matte for reflected light microscopy

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

Skage:

I don't have the answer, but you may want to post this
question on Metallography.com for wider exposure.

Stu Smalinskas, P.E.
Metallurgist
SKF
Plymouth, Michigan

--- shem-at-laurentian.ca wrote:
}
} Hello Everyone,
} Is there anyone out there who has experience etching
} Cu and Ni sulfides, in order
} to see compositional zoning ?
}
} The phases I am interested in are: Cu2S and Ni3-XS2.
} In both these compounds Ni and Cu substitute
} for each other. It is these substitutions which I
} suspect change the etching behaviour of the
} compounds.
} Etching could thus be a quick and dirty alternative
} to excessive X-ray mapping/analysis.
}
} Any input is welcome !
}
} Thanks,
}
} Skage Hem
}
}
}
}
_______________________________________________________________
}
} Skage Hem
} Research Scientist, Ph.D.
} CAF, Department of Earth Sciences
} Laurentian University
} Ramsey Lake Road
} Sudbury, ON
} Canada
} P3E 2C6
}
} ph. 705-675-1151 x4040
} cell. 705-562-7321
} fax 705-675-4898
}
} http://laurentian.ca/geology/Research/hem.html
}
}

__________________________________________________
Do You Yahoo!?
Tired of spam? Yahoo! Mail has the best spam protection around
http://mail.yahoo.com

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5, 20 -- Subject: Re: [Microscopy] Etching of Cu-Ni matte for reflected light microscopy
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From: r-holdford-at-ti.com
Date: Wed, 25 Jan 2006 16:07:49 -0600
Subject: [Microscopy] Texas Society for Microscopy Spring Meeting Announcement & Call for

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

THE TEXAS SOCIETY FOR MICROSCOPY invites you to our Spring 2006 meeting on
April 20-22, 2006 at Alcon Research Labs, 6201 South Freeway, Ft. Worth,
TX 76134

All registration forms and lodging details are available on our web site:
http://www.texasmicroscopy.org/

ABSTRACTS MUST BE RECEIVED BY: March 20, 2006
Advanced Registration Deadline: April 7, 2006.
Advanced registration is strongly suggested to afford TSM an accurate
participant count for event organization.

**Workshops— Thursday, April 20, 2006 (held at Alcon Labs, Ft. Worth)

“Microwave Immunology 101”
Sponsored by Ted Pella, Inc.
Speaker: Rick Giberson, Sr. Applications Engineer


“ESEM: not just for Biology Anymore”
Sponsored by FEI Company
Speaker: Daniel Phifer, Sr. Applications Engineer


**Guest Speaker — Friday, April 21, 2006

“Materials Science in Museums”
Dr. Pamela Vandiver, Professor of Materials Science and Engineering and
Archeology, Co-Director of Program in Heritage Conservation Science at
the University of Arizona, and former Senior Research Scientist at the
Smithsonian Institution’s Center for Materials Research and Education.


--
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
Becky Holdford (r-holdford-at-ti.com)
972-995-2360
972-648-8743 (pager)
SC Packaging FA Development
Texas Instruments, Inc.
Dallas, TX
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~


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From: r-holdford-at-ti.com
Date: Wed, 25 Jan 2006 16:18:42 -0600
Subject: [Microscopy] Call for Papers--Contamination Control in Electron and Ion Microscopy

Contents Retrieved from Microscopy Listserver Archives
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Invitation and Call for Papers
Contamination Control in Electron and Ion Microscopy
Microscopy and Microanalysis 2006, Chicago, IL
Organizers: Ronald Vane (XEI Scientific) and Andrįs E. Vladįr, Ph.D. (NIST)
Session A17 (POSTERS ONLY)
ABSTRACTS DUE February 15, 2006

This session is being organized to present the latest research on
contamination sources and control techniques in charged beam microscopy.
Herein we invite papers on all aspects of contamination control inside
electron microscopes or focused ion beam tools including sources of
contamination, the effects of contamination, contamination artifacts and
interference on microscopy results, and contamination control methods
and techniques. Contamination includes hydrocarbons, particulates, and
other foreign matter that may interfere with imaging and analysis.

Traditionally, contamination control in SEMs has focused on pump oils,
fingerprints, dirty specimens, and good vacuum practice in manufacturing
and service. The use of dry pumps at all stages of the vacuum system of
new FESEMs (Field Emission SEMs) and the use of better vacuum practices
on the part on users and manufacturers have reduced contamination, but
not eliminated problems. Most commonly, tools in the field begin to
exhibit contamination artifacts after several months of use due to trace
amounts of contaminants brought in with specimens. The Semiconductor and
nano-sciences industries have demanded tools that can image structures
{2 nm in size at { 2kV. Instrument manufacturers have responded with
Field Emission tools that easily produce better than 400 kX
magnification at high contrast with low kV beams. Control of
contamination has assumed greater significance as semiconductor
companies move to ever smaller dimensions. It is now common to observe
features with {2 kV and {10 nm in size, close to these tools’ resolution
limits. In such cases, the slightest amount of hydrocarbon (HC)
contamination in the chamber can cause loss of resolution and contrast.
The electron beam reacts with any stray HC in the beam path or on the
surface creating HC ions that condense and form hydrocarbon deposits on
the area being scanned. Even with baking, dry pumps and LN traps,
artifacts and contamination haze remains.

What are the sources and what can be done? Some Research Topic Ideas:

* Do Hydrocarbons adsorbed on materials migrate on the surface to
interact with charged beams to form deposits or is vapor phase transport
more important? The effectiveness of cold fingers and Evactron(R)
chamber cleaning suggests that vapor phase transport of contaminants is
an important alternative mechanism to surface transport.

* Is the gas phase interaction of the electron beam and contaminant
molecules important in SEM contamination buildup? Electron impact
ionization is a common technique in mass spectrometry of organics. What
is the importance of electron impact ionization on vapor phase organics
in the SEM in causing contamination build up in scanned areas?

* What gives better results: plasma cleaning every specimen before
introduction to the SEM chamber, Evactron chamber cleaning, or Evactron
cleaning specimens in the chamber?

* Study Hydrocarbon removal rates for different decontamination methods,
for different types of hydrocarbons and concentrations, under different
Evactron De-Contaminator power and pressure settings, and for different
pump speeds and flow rates during Evactron cleaning.

* How fast does AMC (atmospheric molecular contamination) accumulate on
specimens in different environments after they are cleaned? Freshly
prepared and cleaned specimens are known to be freer from contamination
than specimens that have sat in room air for several days. Quantify this
effect.

* Establish a standard procedure for depositing repeatable amounts of
contamination for removal technique studies.

* Establish a protocol for measuring tool contamination that can be
transferred between tools and used to compare tools or used to establish
contamination standards or limits.


--
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
Becky Holdford (r-holdford-at-ti.com)
972-995-2360
972-648-8743 (pager)
SC Packaging FA Development
Texas Instruments, Inc.
Dallas, TX
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~


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From: kjl226-at-vt.edu
Date: Wed, 25 Jan 2006 19:39:41 -0600
Subject: [Microscopy] viaWWW: Embedding spores

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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: kjl226-at-vt.edu
Name: Kathy

Organization: Virginia Tech

Title-Subject: [Filtered] Embedding spores

Question: What is the best way to process and embed spores?

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

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From: Ken.Blight-at-cancer.org.uk
Date: Thu, 26 Jan 2006 07:43:03 -0600
Subject: [Microscopy]

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

Dear Ralf,

Why not just ask Olympus, I'm pretty sure they will have one as its
still a current microscope. If you are happy with a photocopy expect to get
it free of charge from your local rep (and I'm sure they will email you the
pdf version as well).

Zeiss and Leica now have all the recent microscope manuals on-line, but I
have obtained photocopied manuals for 20+ year old microscopes and fittings,
and once even a BASIC program hardware/software guide for a very odd
programmable HP+Zeiss motorised XY stage discontinued in the 1970's (in
the 1990's). The most interesting bit was the BASIC 'Trace on' debugging
command: 'TRON', but to be honest it worked better when Disney did it.

Keith

----------------------------------------------------------
Dr Keith J Morris
Imaging Facilities Manager
Cell Biology Division
Institute of Ophthalmology
University College London
11-43 Bath Street
London EC1V 9EL

Tel: 020 7608 4050
Fax: 020 7608 4034
email: keith.morris-at-ucl.ac.uk


----- Original Message -----
X-from: {rcommon-at-msu.edu}
To: {keith.morris-at-ucl.ac.uk}
Sent: Wednesday, January 25, 2006 5:27 PM

Hi
I would be very interested in hearing from anyone who has
experience of imaging plates in electron microscopy.

Ken Blight
Senior Scientific Officer
Electron Microscopy
Cancer Research UK
London
England.

==============================Original Headers==============================
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From: joe.p.neilly-at-abbott.com
Date: Thu, 26 Jan 2006 07:59:47 -0600
Subject: [Microscopy] Request for Speakers at Pharmaceutical Symposium , M&M 2006

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

The session organizers for the Pharmaceutical Symposium of M&M 2006 are
looking for additional speakers. Presentations should discuss biological
or material science applications of significance to the pharmaceutical
industry. The meeting will be held in Chicago, July 30 - Aug. 3. The
abstract submission deadline is Feb. 15, 2006. If you are interested in
submitting a platform or poster presentation, instructions for abstract
submission are available at the meeting web site at
http://mm2006.microscopy.org/ . For additional questions please contact
the session chairs at the addresses below. Thanks.

Joe Neilly
Abbott Laboratories
200 Abbott Park Rd.
Bldg. AP31, Dept. R4R9
Abbott Park, IL 60064-6202
email: joe.neilly-at-abbott.com
voice: 847-938-5024

Jim DiOreo
Baxter Healthcare
RL WG3-2S
Route 120 and Wilson Road
Round Lake, IL 60002
email: jim_dioreo-at-baxter.com
voice: 847-270-4676

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From: edelmare-at-muohio.edu
Date: Thu, 26 Jan 2006 09:13:09 -0600
Subject: [Microscopy] Re: [Filtered] AskAMicroscopist: Printer for an SEM

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

Francois:

We have a networked color laser printer in our lab which we
use to print routine image / working print from our SEM's and otehr
microscopes. We print images and sectra, EBDS, etc. What we
have found is that fewer and fewer folks actually want hardcopies.
They want and use electronic versions, since even for publication
nearly all of the journals these days want electronic only. My printer
supplies are about 20% or lower of what they were 2-3 years ago
(i.e. over an 80% decrease in printing - at the same time I've seen a
28% increase in user numbers).

For high quality and archive prints we use our Epson 9600 wide
format printer - we generally use if for posters. I would not hesitate
to get another Epson Pro series, perhaps in a smaller format for
more routine size prints. Or look at the HP highend printers.

But remember the inks will dry in the printers if they are not
regularly used. Some of the manufacturers have inks which are
designed to last longer in the printer - but they are not the low- to
medium end printers. Dry inks like laser printers and wax/polymer
"phaser" printers have an advantage here.

FYI: We have a Lexmark C752 color laser printer. And whereas
we've been very very happy with the durablity and quality of the
Lexmark Black / Greyscale laser printers, we have not been thrilled
with the image quality of the C752 prints, and yes we use the high
quality color laser paper.


On 25 Jan 2006, at 7:44, fmalherbe-at-swin.edu.au wrote:

}
}
}
} ----------------------------------------------------------------------
} ------ The Microscopy ListServer -- CoSponsor: The Microscopy Society
} of America To Subscribe/Unsubscribe --
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} Below is the result of your feedback form (NJZFM-ultra-55). It was
} submitted by (fmalherbe-at-swin.edu.au) from
} http://www.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html
} on Tuesday, January 24, 2006 at 18:23:45
} ----------------------------------------------------------------------
} -----
}
} Email: fmalherbe-at-swin.edu.au
} Name: Francois Malherbe
}
} Organization: Swinburne University of Technology
}
} Education: Graduate College
}
} Location: Melbourne, Victoria, Australia
}
} Question: We are purchasing a new SEM, what are the basic specs you
} would recommend for the printer:
}
} - inkjet, laser or other?
} - USB, serial?
} - dpi?
}
} Thank you for your help.
}
} ----------------------------------------------------------------------
} -----
}
} ==============================Original
} Headers============================== 9, 12 -- From
} zaluzec-at-ultra5.microscopy.com Wed Jan 25 07:33:17 2006 9, 12 --
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} Subject: [Filtered] AskAMicroscopist: Printer for an SEM 9, 12 --
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Richard E. Edelmann, Ph.D.
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

"WE ARE MICROSOFT.
RESISTANCE IS FUTILE.
YOU WILL BE ASSIMILATED."

==============================Original Headers==============================
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From: mcauliff-at-umdnj.edu
Date: Thu, 26 Jan 2006 09:19:09 -0600
Subject: [Microscopy] Re: plates

Contents Retrieved from Microscopy Listserver Archives
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Do you mean using glass plates instead of film? If so .....
Plates are much thicker and heavier, these factors contribute to storage
problems.
Plates break when dropped, film does not.
I know people who insist that plates give a better image but I have seen
no evidence of that.
The boxes that Kodak plates came in were great for storing lantern slides.

Geoff

Ken.Blight-at-cancer.org.uk wrote:

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From: Ken.Blight-at-cancer.org.uk
Date: Thu, 26 Jan 2006 09:29:17 -0600
Subject: [Microscopy] Imaging plates

Contents Retrieved from Microscopy Listserver Archives
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Sorry for the confusion caused -The imaging plates I am interested in
are electron detection plates for high quality digital images. I have
no other details otherwise I would not be asking the question!


ken

Ken Blight
Senior Scientific Officer
Electron Microscopy
Cancer Research UK
London
England.

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From: mike.reedy-at-cellbio.duke.edu
Date: Thu, 26 Jan 2006 11:06:05 -0600
Subject: [Microscopy] Re: Imaging plates

Contents Retrieved from Microscopy Listserver Archives
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What you want is described at
http://www.berthold.com.au/imaging_pages/FDL-5000.html

and at
http://www.ditabis.de/

I have no experience with use of these in EM. An impressive 3D
cryo-EM paper based on use of the imaging plates was published by
Holmes et al in
Nature (2003), 425:423-427.
Use of imaging plates requires drying the media before loading into
the camera, and requires breaking camera vacuum and accessing the
camera in order to remove media to "develop" (scan & digitize) the
recorded images and regenerate the plate. Both steps are avoided
when using CCD display and recording of EM images, which provide the
other route for directly digitizing EM images without processing and
scanning photographic negatives.

-mike reedy-


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Michael K. Reedy, M.D.
Duke Univ. Med. Center
Dept. Cell Biology, Box 3011 (for U.S. Mail)
458 Alex Sands Bldg, Research Dr (courier)
Durham, NC 27710

Office 919-668-2534
Lab 919-684-5674
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From: beth-at-plantbio.uga.edu
Date: Thu, 26 Jan 2006 13:42:00 -0600
Subject: [Microscopy] x-ray film processors

Contents Retrieved from Microscopy Listserver Archives
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I hope you all will forgive this detour from microscopy.
If you have a x-ray film processor in your lab and would care to
comment on X-ray film processors I would love to hear from you
off-list. We have an AFP Medical-Mini X-ray film processing system but
we are looking into replacing it with a Konica SRX-101A desktop
processor.
Advice/ opinions on this equipment would be greatly appreciated.
Thanks,
Beth

**********************************************************************
Beth Richardson
EM Lab Coordinator
Plant Biology Department
University of Georgia
Athens, GA 30602-7271

Phone - (706) 542-1790 & FAX - (706) 542-1805
http://www.plantbio.uga.edu/emlab

"Between the two evils,
I always pick the one I never tried before". Mae West (1893-1980)
*******************************************************************

"And it's only the giving that makes you what you are".
Wond'ring Aloud, Jethro Tull (Aqualung)

************************************************************************
***



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From: info-at-lheritier-sa.com
Date: Fri, 27 Jan 2006 08:03:06 -0600
Subject: [Microscopy] viaWWW: TEM - Seeking to acquire JEOL jem100c or equivalent

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Email: info-at-lheritier-sa.com
Name: Patrice MENARD

Organization: LHERITIER SAS

Title-Subject: [Filtered] TEM - Seeking to acquire JEOL jem100c or equivalent.

Question: We are a small French company based in the Paris region, currently looking to replace our JEOL jem100c transmission electron microscope, which unfortunately, has broken down.

We need to acquire a second-hand microscope similar to the model we actually have but as we wish to use it mainly as an electron generator, to test the performance of the customized cameras we manufacture, we would accept a tem microscope, which does not offer all the requirements necessary for good quality imagery.

If you have such equipment and are willing to give us the opportunity of acquiring it, please contact us via our e-mail address: info-at-lheritier-sa.com.


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From: wesaia-at-iastate.edu
Date: Fri, 27 Jan 2006 13:37:35 -0600
Subject: [Microscopy] Need ISIS power supply

Contents Retrieved from Microscopy Listserver Archives
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It looks like we are losing the +24V section of the power supply on our
Oxford ISIS EDS. Does anyone have a spare power supply, or even an
entire ISIS unit they would be willing to part with? It probably should
be a model 200 or 300.

Please contact me off-line and we can discuss the details.

Warren Straszheim
Materials Analysis and Research Lab
Iowa State University
515-294-8187


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From: raynald.gauvin-at-mcgill.ca
Date: Fri, 27 Jan 2006 15:45:24 -0600
Subject: [Microscopy] viaWWW: Advanced Techniques in Microscopy for Materials

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Email: raynald.gauvin-at-mcgill.ca
Name: Raynald Gauvin

Organization: McGill University

Title-Subject: [Filtered] Worshop

Question: Title: Advanced Techniques in Microscopy for Materials
Characterization

Lecturers: David Joy, University of Tennessee
Eric Lifshin, State University of New York
George Vander Voort, Buehler Ltd.
Raynald Gauvin, McGill University
Brendan Griffin, University of Western Australia
Rocco Cerchiara, Fischione Instruments
Pierre Hovington, Hydro-Quebec Research Institute
Tom Kelly, Imago Scientific Instruments
Scott Sitzman, HKL Technology
Marin Lagace, Hydro-Quebec Research Institute

When: May 8-12, 2006

Where: McGill University, Department of Mining, Metals and Materials
Engineering
M.H.Wong Building
3610 University Street
Montreal, Quebec, Canada
H3A 2B2

Contact: Prof. Raynald Gauvin
Tel: (514) 398-8951
Fax: (514) 398-4492
E-mail: raynald.gauvin-at-mcgill.ca


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From: zaluzec-at-microscopy.com
Date: Mon, 30 Jan 2006 08:31:30 -0600
Subject: [Microscopy] MM 2006 Paper Submission Deadline - Feb 15, 2006

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Colleagues

The worlds largest and most comprehensive microscopy and microanalysis meeting is coming to Chicago July 30-Aug 3, 2006. This is the perfect opportunity for both your and as well as undergraduate/graduate students to gain valuable insights to recent work by interacting with the worlds most prominent researchers, as well as learn about the latest techniques in the scientific symposia and in instrumentation.

In addition to more than 50 scientific sessions and symposia there will be a number of short courses/tutorials starting on the weekend and extending through the week.

The meeting also hosts the largest commerical exhibition of microscopy instrumentation at any venue.

This will be the perfect opportunity for you to take advantage of the premier international meeting happening in the USA as well as make valuable contacts for the future.


Of particular importance to students is the availability of scholarship, and other funding opportunities for students which will allow you to attend the meeting at reduced costs.

*Student receive reduced registration rates
* Scholarships and Awards are available from the sponsoring societies
* Poster awards are presented for the best student contributions
* Student Bursaries (i.e. work part time during the meeting) can be used to help defray your expenses

A limited number of awards are also available for professional technical staff.

To be eligible for these opportunities you must submit a paper for either platform or poster presentation at the meeting.
The deadline for submitting a paper to MM2006 is Feb. 15, 2006 (there are no extensions), which is only a few weeks away.

If you have any question please feel free to contact any of the Organizers, for detailed information please visit the meeting WWW site
(http://mm2006.microscopy.org).

Again the paper submission deadline is Feb 15, 2006. I will also point out that the submission process is now entirely electronic so you still have plenty of time to prepare your work and submit it for review and consideration. All papers for accepted for the meeting are included in the proceedings which is published as supplement to the Journal of the Microscopy Society of America - Microscopy & Microanalysis.

Disclaimer: In additions to being your Friendly Neighborhood SysOp, I have the "honor" (cough) of being the Co-chair of the local arrangements committee for this meeting, thus I have a vested interest in getting you to come to Chicago!


Cheers...

Nestor
Your Friendly Neighborhood SysOp

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From: pjoshi-at-miami.edu
Date: Mon, 30 Jan 2006 08:58:43 -0600
Subject: [Microscopy] Re: Looking for EHT X Regulator unit for Phillips EM 300 TEM

Contents Retrieved from Microscopy Listserver Archives
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Hi Everyone
I am looking for a EHT X-regulator for our old Phillips EM 300 TEM. Anybody knows a good place to buy this part or have such parts on sale please contact me at pjoshi-at-miami.edu
Thank you

Pratik P. Joshi, Ph.D.

Assistant Scientist / Lab Manager
Center for Advanced Microscopy
Department of Chemistry, University of Miami
1301 Memorial Drive, Room 315
Miami, FL- 33146
Ph: (305)284-4736, Fax: (305)284-1880


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From: brad.huggins-at-bp.com
Date: Mon, 30 Jan 2006 20:09:45 -0600
Subject: [Microscopy] viaWWW: LM, Anyone with "Digital Microscope"

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Email: brad.huggins-at-bp.com
Name: Brad Huggins

Organization: BP Chemicals, Naperville, IL

Title-Subject: [Filtered] LM, Anyone with "Digital Microscope" familiarity/experience?

Question: (trouble with MS Outlook and spam errors, so I'm using this format)

For several remote, light microscope applications, I have been looking at the Keyence VHX-100, Digital Microscope. It looks to be a very capable and versatile tool. Aside from its portability, 3D imaging capability appears quite impressive to me. It looks capable of adding - very significantly - to the limited depth-of-focus imaging condition of our existing light microscope systems.

Does anyone on the ListServer have any experience (good, bad or otherwise) with this microscope? Or similar digital microscopes? If you would rather not respond online, please share with me off-line. I will share a summary of "anonymous responses" as
appropriate.

Thanks in advance,

Brad Huggins

BP Chemicals
Naperville, IL
brad.huggins-at-bp.com
630 420-3668


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From: aarti_harle-at-yahoo.co.in
Date: Tue, 31 Jan 2006 06:19:06 -0600
Subject: [Microscopy] mounting media for fluorescence/ confocal microscope

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Dear All

Does anyone has experience with 'Vectashield with or
without DAPI' (Vecotor Labortories) antifade mounting
media for confocal microscopy

Regards
Shrunali Kulkarni
Scientist
Institute of Microbial Technology
India

__________________________________________________
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From: aarti_harle-at-yahoo.co.in
Date: Tue, 31 Jan 2006 22:35:21 -0600
Subject: [Microscopy] Sample preparation technique for confocal microscopy

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Dear All

I would like hear the comment on various sample
preparation technique for confocal microscopy
(indirect Immunofluorescence)
which gives 1) best presarvation
2) minimum background noise

Is it advisible to dry the slide before switching over
to 1 Ab and 2Ab. I mean to ask is this drying in
between lead to some form of precipitate???

thanks in advance

Regards,
Shrunali Kulkarni
Scientist
Institute of Microbial Technology
India

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From: hagglundk1-at-nku.edu
Date: Wed, 1 Feb 2006 08:09:03 -0600
Subject: [Microscopy] ESEM chamber gas choices

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Dear Shrunali

Yes it works fine (as do many anti-fadents e.g Citifluor
http://www.citifluor.co.uk ), although we rarely get problems with DAPI
bleaching - I won't mention Hoescht as I can never spell it. Normally the
likes of Cy5 or TRITC type fluorochromes bleach first, and anyway DAPI is
normally only there in our case to identify the animal cells more easily
(and it really makes specimen focussing a cinch with our 'low contrast'
samples). Vectashield antifadents stop samples fading really quickly rather
than eliminating fading. So you still have to minimise laser (and Hg lamp)
exposure times (e.g. increase scan speed, reduce image size & image
averaging, increase gain, and, normally in the last resort, open up the
confocal iris), particularly if you are doing Z stacks (naturally time-lapse
won't be a problem with fixed 'Vectashield' samples).

Being Cell Biology, most of our specimens are derived from cell cultures and
so have little contrast, making cell visual separation and location more
difficult even with DIC. We only have a little 5 Mw 'violet' 405nm laser
with our Leica SP2 AOBS confocal, but it works very well with nuclear stains
despite being at the very edge of the DAPI/Hoescht excitation spectrum. We
mostly use Mattek dishes (Petri dishes with a hole at the base and little
cover slips glued on - see http://www.glass-bottom-dishes.com/ ) as all our
microscopes are inverted and we only use high power oil objectives. They
have the advantage that we can simply drip the Vectashield into the Petri
dish on top of the fixed cells and replenish if it dries out (samples often
keep for days or longer in a darkened fridge). With glass slides, the
coverslip needs sealing on over the Vectashield with nail varnish, but the
nail varnish (and marker pen ink) often dissolve into the immersion oil
making a bit of a mess, particularly with the inverted microscopes we use.
The nail varnish also sticks to the stage slide holder (as the slide has to
be placed upside down in inverted systems) making a sticky mess and knocking
the sample out of level.

With regard to preparation, we don't deal with microbiological specimens,
just mammalian cells - so I won't offer any advise. There are plenty of
recipes on the internet and in books though, and you can try contacting
staff at another microbiological institutes via on-line searches or
traditional 'old boy' networks. This list server tends to be biased a little
towards electron microscopy. You reduce confocal image 'noise' in samples by
lowering the scan speed, increasing image averaging and upping laser power
on the confocal - the exact opposite of that required to minimise beaching.
So there is always a compromise between image quality and fluorochrome
bleaching. If the sample preparation has worked well your sample would be
expected to be quite bright initially (prior to bleaching) - often problems
with dark images are due to experimental failure rather than the confocal
microscope and it's software.

Regards

Keith

----------------------------------------------------------
Dr Keith J Morris
Imaging Facilities Manager
Cell Biology Division
Institute of Ophthalmology
University College London
11-43 Bath Street
London EC1V 9EL

Tel: 020 7608 4050
Fax: 020 7608 4034
email: keith.morris-at-ucl.ac.uk



----- Original Message -----
X-from: {aarti_harle-at-yahoo.co.in}
To: {keith.morris-at-ucl.ac.uk}
Sent: Tuesday, January 31, 2006 1:02 PM

Dear Shrunali,

I forgot to add it (although I expect you have the link already) : The full
details on Vectorshield products [e.g. Hardset and Mounting Medium versions]
can be found on the manufacturers website :

http://www.vectorlabs.com/products.asp?catID=279&locID=609338

It naturally has the 'test on an inconspicuous area before using'
disclaimer. Anti-fadents have been reported to occasionally cause a leaching
of stain and loss of fluorescence in some instances.

Regards

Keith

----------------------------------------------------------
Dr Keith J Morris
Imaging Facilities Manager
Cell Biology Division
Institute of Ophthalmology
University College London
11-43 Bath Street
London EC1V 9EL

Tel: 020 7608 4050
Fax: 020 7608 4034
email: keith.morris-at-ucl.ac.uk


----- Original Message -----
X-from: {aarti_harle-at-yahoo.co.in}
To: {keith.morris-at-ucl.ac.uk}
Sent: Tuesday, January 31, 2006 1:02 PM

I am interested in hearing what type of gases people are using in their
ESEM applications. We recently learned our silicon drift detector is
incompatible with water vapor, and we apparently cannot complete any
x-ray microanalysis while working in standard ESEM or VP modes (I was
pretty surprised to learn this). We are thinking that using a dry gas
such as nitrogen or helium will allow us to work in ESEM modes and use
our x-ray system as well, as there is no potential for moisture
condensing on the crystal surface.

Our ESEM is a Quanta 200, and it is not equipped with the peltier stage,
so we mainly use the environmental modes to alleviate charging in
uncoated samples. The x-ray system was purchased with the microscope
about four years ago. I would rather not discuss the name of the x-ray
vendor on the list, as we have a significant investment in this detector
and do not foresee finding the money to purchase a new one soon.

Our first concern is the impact of the gas on spectra. We can coat
samples and run them in high vacuum mode, but customers like spectra
that represent the sample and not the coating material. We periodically
have samples that cannot be coated, so ESEM becomes critical for our
imaging needs. If the gas is going to have a dramatic impact on signal,
are we better off coating and running high vacuum?

Is there a best choice for chamber gas? Our service engineer recommends
nitrogen as having benefits for keeping the system clean. We can set it
up fairly quickly, and I have a spare tank and regulator. I have heard
of people using helium and believe that I read somewhere that there was
some advantage to using helium.

I am still trying to get information from the vendor on whether this
will allow us to use the x-ray and ESEM simultaneously, or whether we
have options with different collimators or windows. Their responses
have been pretty slow coming.



Karl Hagglund
Biological Sciences, SC-102B
Northern Kentucky University, Nunn Drive
Highland Heights, KY 41099
859-572-5238


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From: lesley.bechtold-at-jax.org
Date: Wed, 1 Feb 2006 09:23:30 -0600
Subject: [Microscopy] Seeking information

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Hi,

I am looking for information about staffing, equipment and general practices in microscopy facilities at other institutions. If anyone else is interested in the same sort of information, please reply to me directly. Thank you.

Lesley Bechtold


Lesley S. Bechtold
Senior Manager, Histopathology & Microscopy Sciences
The Jackson Laboratory
600 Main St.
Bar Harbor, ME 04609
207-288-6322



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From: ptomic-at-ciclonsemi.com
Date: Wed, 1 Feb 2006 10:00:46 -0600
Subject: [Microscopy] viaWWW: Electrolytical Metal Analysis Tool

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Email: ptomic-at-ciclonsemi.com
Name: Peter Tomic

Organization: Ciclon Semiconductor Device Corp.

Title-Subject: [Filtered] ELYMA

Question: I would like to hear from people in the semiconductor industry on their experience with ELYMAT [Electrolytical Metal Analysis Tool]. This is an imaging system for the analysis of metal and oxygen contamination in Si wafer technology.

Peter Tomic
Ciclon Semiconductor Device Corp.

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From: twigg-at-estd.nrl.navy.mil
Date: Wed, 1 Feb 2006 10:08:11 -0600
Subject: [Microscopy] viaWWW: Postdoctural Position at NRL

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Email: twigg-at-estd.nrl.navy.mil
Name: Mark Twigg

Organization: Naval Research Laboratory

Title-Subject: [Filtered] Postdoctural Position

Question: Postdoctoral Research Position
Electronics Science and Technology Division
Naval Research Laboratory, Washington, DC

We are seeking a Postdoctoral fellow with a strong transmission electron microscopy and extended-defects/nanostructures background to join our electronic materials program at NRL. You will utilize your skills to characterize SiC and GaN epitaxial layers, as well as nanoelectronic materials based on colloidal gold. We have our own Hitachi H9000UHR HRTEM and sample preparation laboratory. We also have access to a Philips CM30, a JEOL 2200 FETEM with Omega energy filter and Z-contrast STEM, and an FEI Nova 600 Dual Beam FIB. The qualified candidate should have a Ph.D. in Materials Science, Physics, Chemistry, or a related physical science or engineering discipline. Experience in a variety of electron microscopy techniques including HRTEM, CBED, and EDXS, and EELS is important. Experience with FIB, XTEM, and mechanical lapping sample preparation techniques is also desirable.
Participants must be citizens or permanent residents of the United States. Two postdoctoral study programs are available at the Naval Research Laboratory: NRC and ASEE. NRC postdoctoral positions are administered by the National Research Council, a division of the National Academy of Sciences. The ASEE Postdoctoral Fellowship Program is administered by the American Society for Engineering Education. Further information about NRL's NRC post-doc program is found on the NRC web site (http://hroffice.nrl.navy.mil/jobs/postdoc.htm).
Fellowships are awarded for one year and may be extended for a second year. The base annual stipend for both programs the first year is $62,886.

Point of Contact: Dr. Mark E. Twigg
Naval Research Laboratory
Code 6812
4555 Overlook Avenue, S.W.
Washington, DC 20375-5320

Email: twigg-at-estd.nrl.navy.mil
tel: (202) 404-8543
fax: (202) 404-7194


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From: dyel-at-mail.nih.gov
Date: Wed, 1 Feb 2006 11:14:32 -0600
Subject: [Microscopy] viaWWW: Tyrodes-Cacodylate Buffer

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Email: dyel-at-mail.nih.gov
Name: Chip Dye

Organization: NIH-NICHD

Title-Subject: [Filtered] Tyrodes-Cacodylate Buffer

Question: Dear ListServer:

Has anyone used this buffer before? I plan on using this to make up a fixative to perfuse fix a rat. I will be looking at myelin of the sciatic nerve. Why does this buffer have such a high osmolarity? Any thoughts?

Thank you!

Tyrodes-Cacodylate Buffer (mammalian Ringers) (401 mOsmols)
1 liter dd H2O
NaCl 6 gm
KCl 0.2 gm
Na(CH3)2AsO2.3H2O 10.7 gm
MgCl2.6H2O 0.1 gm
NaHCO3 1.0 gm
Glucose 1.0 gm
CaCl2.2H2 0.2 gm
(adjust to pH 7.4 with 3.1 %HNO3)


Chip Dye

Microscopist
Microscopy & Imaging Core
NICHD, NIH
Bldg. 49, Room 5W14
Bethesda, Maryland 20892-4480
Phone: 301-496-3627
FAX: 301-496-9939
Email: dyel-at-mail.nih.gov
Pager : Dial 102, 11568, your phone number
Outside campus: Dial 1-800-NIH-BEEP, 11568, you phone number
http://mic.nichd.nih.gov


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From: lcgould-at-med.cornell.edu
Date: Wed, 1 Feb 2006 12:54:15 -0600
Subject: [Microscopy] Re: viaWWW: Tyrodes-Cacodylate Buffer

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Chip,
I've never used Tyrode's in conjunction with cacodylate. Years ago,
when the lab I was in was collaborating with an electrophysiologist
looking at structure-function relationships in heart muscle, we used
Tyrode's because it is a bicarbonate-buffered solution and he could
keep the isolated papillary muscles happy in it for hours as long as
he bubbled oxygen-CO2 through it. Lovely structure (see various
papers by Robinson, TR, etal during the 1980's).
Lee
--
Leona Cohen-Gould, M.S.
Sr. Staff Associate
Director, Electron Microscopy & Histology Core Facility
Manager, Optical Microscopy Core Facility
Joan & Sanford I. Weill Medical College
of Cornell University
voice (212)746-6146
fax (212)746-8175
http://www.cornellcelldevbiology.org

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From: dyel-at-mail.nih.gov
Date: Wed, 1 Feb 2006 13:03:24 -0600
Subject: [Microscopy] viaWWW: Tyrodes-Cacodylate Buffer

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Email: dyel-at-mail.nih.gov
Name: Chip Dye

Organization: NIH-NICHD

Title-Subject: [Filtered] Tyrodes-Cacodylate Buffer

Question: Dear ListServer:

Has anyone used this buffer before? I plan on using this to make up a fixative to perfuse fix a rat. I will be looking at myelin of the sciatic nerve. Why does this buffer have such a high osmolarity? Any thoughts?

Thank you!

Tyrodes-Cacodylate Buffer (mammalian Ringers) (401 mOsmols)
1 liter dd H2O
NaCl 6 gm
KCl 0.2 gm
Na(CH3)2AsO2.3H2O 10.7 gm
MgCl2.6H2O 0.1 gm
NaHCO3 1.0 gm
Glucose 1.0 gm
CaCl2.2H2 0.2 gm
(adjust to pH 7.4 with 3.1 %HNO3)


Chip Dye

Microscopist
Microscopy & Imaging Core
NICHD, NIH
Bldg. 49, Room 5W14
Bethesda, Maryland 20892-4480
Phone: 301-496-3627
FAX: 301-496-9939
Email: dyel-at-mail.nih.gov
Pager : Dial 102, 11568, your phone number
Outside campus: Dial 1-800-NIH-BEEP, 11568, you phone number
http://mic.nichd.nih.gov


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From: jae5-at-lehigh.edu
Date: Wed, 1 Feb 2006 15:13:05 -0600
Subject: [Microscopy] Free TEM Workshop Materials and Geology

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Pan American Advanced Studies Institute
on
Transmission Electron Microscopy in Materials Science

July 9 to July 22, 2006

Expenses paid
Further information: http://www.pasi-tem2006.cl

At the University of Chile - in Santiago, Chile - a new field-emission
TEM is being installed. This microscope will form the basis of a Pan
American Advanced Studies Institute which will cover principles and
applications of TEM to topics in materials and geological sciences.
The Institute is funded by the National Science Foundation of the United
States.

We invite applications from students and young researchers who wish to
participate in this two-week intensive workshop. APPLICANTS WHO ARE
ACCEPTED WILL HAVE THEIR EXPENSES PAID. The number of participants in
the workshop will be limited to 48.

The lectures and laboratories (presented by a distinguished group of
lecturers) will treat a wide range of topics: principles and
applications of transmission electron microscopy; microanalysis; latest
results and advances; and sample preparation.

The Institute is open to participants from any country in the Americas
(including the U.S.A.). Participation by women and members of minority
groups is greatly encouraged. Details of how to apply are given on the
web site:
http://www.pasi-tem2006.cl
The closing date for applications is March 13, 2006

--
...........
Alwyn Eades
Department of Materials Science and Engineering
Lehigh University
5 East Packer Avenue
Bethlehem
Pennsylvania 18015-3195
Phone 610 758 4231
Fax 610 758 4244
jae5-at-lehigh.edu


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From: gary-at-gaugler.com
Date: Wed, 1 Feb 2006 20:27:17 -0600
Subject: [Microscopy] Re: ESEM chamber gas choices

Contents Retrieved from Microscopy Listserver Archives
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The Quanta 200 uses a W filament. Unless it is a SFEG.
If there are no ion pumps, you should be able to use
Nitrogen or He. If there is an ion pump, do not use He.
It will kill the pump.

I found that for VP (20-120Pa) that N2 works fine and is
better than air (less vacuum issues). So, for ESEM, I would
think that N2 ought to be the gas of choice as well.

For VP mode, I do quant with EDAX Genesis using collected
values at two pressure/vacuum values. This is their VIP
option. I find good correlation between it and high vacuum
mode. Which EDS are you using? Does it have an ESEM
option mode?

gary g.




At 06:45 AM 2/1/2006, you wrote:



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From: W.Muss-at-salk.at
Date: Thu, 2 Feb 2006 09:16:29 -0600
Subject: [Microscopy] Re: Tyrodes-Cacodylate Buffer

Contents Retrieved from Microscopy Listserver Archives
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Good morning,
dear Chip,

it is a long time ago I have used a Tyrode-GA-mixture as a perfusion
fixative for rat brain (selective hypothalamic target preparation
containing ventricle III as well as magnocellular nuclear regions,
providing "open" blood vessels/capillaries in that region; followed by a
3-dim reconstruction of those nuclear regions)....

Later on I had to use also such perfusion mixtures for retrograde perfusion
of pig (renal) arteries......
I don't know or have at hand now the original receipt of } Tyrode {'s
Mammalian Ringer solution, but have found in my methods/techniques
collection } old { descriptions and protocols of the respective solutions and
measurement data.

I have seen that your mixing formula varies only in the amount of
Glucose......you indicate: 1 gm, in my descriptions I do have listed 10
grms.

On the other hand, when working with Tyrode's, initially I haven't mixed
the buffer with sodium cacodylate, but instead with a very small amount of
sodium-dihydrogenphosphate as follows:
NaCl: 6.0 gm
KCl: 0.2 gm
CaCl2 (as CaCl2.2H2O) 0.2 (0.25g) gm
MgCl2 (as MgCl2.6H2O) 0.1 (0.2. g) gm
sodium-dihydrogenphosphate 0.05 gm
Sodium-hydrogencarbonate(NaHCO3) 1.0 gm
Glucose 10.0 gm
------------------------------------------------------------------------
----
ad 1000 ml A.bidest (DD A.dest.)
Phosphate(s) must be dissolved extra and should finally be added when all
other } powder-substances { (! especially CaCl2 and MgCl2) are readily
dissolved.

I have noted in the protocol:
initial pH of the resulting solution: 8.4, approx. 275-277 mosmol
(measuring also pH 8.14, mosmol: 310, 290, 285, 282, 295, 295).

pH-correction with approx. 2.95 ml 1 N HCl to pH. 7.2

The fixative for perfusion consisted of:
160 ml 25% glutaraldehyde (which means a final GA-concentration of 4% in
the fixative) to be mixed with the above mentioned Tyrode stock solution to
an endvolume of 1000 ml.
If there formed a pale/ milky precipitate (which sometimes occured, perhaps
due to a poor GA-quality with a high amount of aldehyde polymers - we had
at that time in the late 1970ies - but most probably due to the
Na2HPO4/NaH2PO4 !) the solution should be / has to be filtrated.

After doing so, I noted: pH ==} 8.4, 545-550 mosmol

Since the pH of that fix-mixture increased (????) to 8.4 with time again,
it was necessary to adjust again with some ml of 1 N HCl to a pH of at
least 7.7 (the solution now seemed to act quite well as a } buffer { system !
) and finally,
having adjusted the solution to pH 7.2 again, I then measured 1160 /1190 /
1200 mosmol.

As the following washing buffer I used at that time a Tyrode solution as
given above, but added Glucose 25 g ad 1000 ml solution (which perhaps was
wrong), and measured (after pH correction with approx. 1.3 ml of 1 N HCl to
pH 7.2) 360 / 370 / 375 mosmol.

The 4% glutaraldehyde perhaps will contribute approx. 280 mosmol to the
total osmolarity (if calculating from some own measurements and some tables
found in the literaturefor 1, 1.5, 2.0 and 5 % GA-Solutions, respectively),
the rest up to 1200 mosmol therefore must originate from the (more
complete) dissociation of total ions dissolved in the tyrode's solution.

You state that the Tyrode solution you measured had 400/401 mosmol and is
thought to be unusually high.

I assume that there is a contribution by the sodium-cacodylate you added
(which is, if you use 10.7 g / 1000 ml, a final molarity of 0.05 mol) which
could be in the range of 100 mosmol.

Consider also that due to more effective dissociation of ionic components
in the solution the more diluted your fluid is, osmolarity will increase -
not in a 1:1 mode
(eg. Na-phosphate ions in Na2HPO4 and NaH2PO4 0,1 M will not have double
the osmol amount of 0.05 M, which in fact will be higher due to more
effective dissociation).

In my files I have other/additional data concerning the use of Tyrode's
ringer solution, I you like, I could share those with you on request
offline.

Best regards,

Wolfgang Muss
OR Dr. Wolfgang Muss
EM-Lab
====} with pride: 25 years in operation by 2nd of Feb. 2006 {====
Institute of Pathology, SALK
(Salzburger Landeskliniken gemeinnuetzige GesmbH)
Muellner Hauptstrasse 48
A-5020 SALZBURG AUSTRIA/EUROPE

and/or/alternatively

Paracelsus Medical Private University (PMU)
Institute of Pathology
Electron Microscopy Lab
Muellner Hauptstrasse 48
A-5020 SALZBURG, Austria/Europe
Phone work: +43+662+4482+4720
Mobile phone work:+43+662+4482-57704
Fax-No. at work: ++43+662+4482-882 ext (please, only by indicating: "c/o
W.Muss")
E-Mail work: W.Muss-at-SALK.at

Mobile-phone private: ++43+676+5 369-456
E-Mail private: wij.Muss-at-aon.at
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----------
Von: dyel-at-mail.nih.gov[SMTP:dyel-at-mail.nih.gov]
Antwort an: dyel-at-mail.nih.gov
Gesendet: Mittwoch, 01. Februar 2006 20:07
An: W.Muss-at-salk.at
Betreff: [Microscopy] Tyrodes-Cacodylate Buffer

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Email: dyel-at-mail.nih.gov
Name: Chip Dye
Organization: NIH-NICHD
Title-Subject: [Filtered] Tyrodes-Cacodylate Buffer

Question: Dear ListServer:

Has anyone used this buffer before? I plan on using this to make up a
fixative to perfuse fix a rat. I will be looking at myelin of the sciatic
nerve. Why does this buffer have such a high osmolarity? Any thoughts?

Thank you!

Tyrodes-Cacodylate Buffer (mammalian Ringers) (401 mOsmols)
1 liter dd H2O
NaCl 6.0 gm
KCl 0.2 gm
Na(CH3)2AsO2.3H2O 10.7 gm
MgCl2.6H2O 0.1 gm
NaHCO3 1.0 gm
Glucose 1.0 gm
CaCl2.2H2 0.2 gm
(adjust to pH 7.4 with 3.1 %HNO3)


Chip Dye

Microscopist
Microscopy & Imaging Core
NICHD, NIH
Bldg. 49, Room 5W14
Bethesda, Maryland 20892-4480
Phone: 301-496-3627
FAX: 301-496-9939
Email: dyel-at-mail.nih.gov
Pager : Dial 102, 11568, your phone number
Outside campus: Dial 1-800-NIH-BEEP, 11568, you phone number
http://mic.nichd.nih.gov


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From: jae5-at-lehigh.edu
Date: Thu, 2 Feb 2006 09:36:47 -0600
Subject: [Microscopy] Re: ESEM chamber gas choices

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Water in an ESEM affecting EDS detectors?

There are some things that are not clear about this problem. The
question was written as if the use of water in an ESEM is a problem
specifically for silicon drift detectors. Surely the problem is the
same for all types of detector; the problem is not with the detector but
with the window. In a system working correctly, the vacuum of the
detector is quite separate from the vacuum of the chamber. So it does
not matter what gas you use in the ESEM as long as the window is intact.

Does water in an ESEM cause the window of the detector to develop holes
more quickly than, say, nitrogen in the ESEM? This could be the case,
depending on the window design. We did have window failures in our
ESEM, but we are told that this was a temporary problem and that the
windows being sold now are just fine in an ESEM using water. Does
anyone have specific knowledge on this?

Alwyn Eades

--
...........
Alwyn Eades
Department of Materials Science and Engineering
Lehigh University
5 East Packer Avenue
Bethlehem
Pennsylvania 18015-3195
Phone 610 758 4231
Fax 610 758 4244
jae5-at-lehigh.edu


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From: phillipst-at-missouri.edu
Date: Thu, 2 Feb 2006 11:40:15 -0600
Subject: [Microscopy] Cytoviva system

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Does anyone have experience with the Cytoviva imaging system. It looks to
me (from their website at www.cytovita.com like it is a retrofit
condenser. They claim a resolution better than 50 nm. In fact, their
website says "The typical optical microscope provides a maximum theoretical
resolution limit of 240nm. With resolution below 100nm and detection below
50nm, CytoViva allows you to see details never before possible with
traditional microscopy. " Can someone explain to me how one can improve on
the maximum theoretical resolution?

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



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From: W.Muss-at-salk.at
Date: Thu, 2 Feb 2006 12:40:31 -0600
Subject: [Microscopy] Re: Cytoviva system

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Dear Prof Phillips,
perhaps the instrument of Cytoviva ( http://www.cytoviva.com ) is a further
development of a technique I found in PNAS 2002 (if you want I can provide
you with a pdf of this article). Unfortunately I have not followed that
thread.....

FYI: Title of work:

Fast 100-nm resolution three-dimensional microscope reveals structural
plasticity of mitochondria in live yeast
Alexander Egner, Stefan Jakobs, and Stefan W. Hell*

3370-3375 PNAS March 19, 2002 vol. 99 no. 6
By introducing beam-scanning multifocal multiphoton 4Pi-confocal
microscopy, we have attained fast fluorescence imaging of live
cells with axial super resolution. Rapid scanning of up to 64 pairs
of interfering high-angle fields and subsequent confocal detection
enabled us to perform three to five times finer optical sectioning
than confocal microscopy. In conjunction with nonlinear image
restoration, we demonstrate, to our knowledge for the first time,
three-dimensional imaging of live eukaryotic cells at an equilateral
resolution of ~ 100 nm. This imaging mode allowed us to reveal the
morphology and size of the green fluorescent protein-labeled
mitochondrial compartment of live Saccharomyces cerevisiae (bakers'
yeast) growing on different carbon sources. Our studies show
that mitochondria of cells grown on medium containing glycerol as
the only carbon source, as opposed to glucose-grown cells, exhibit
a strongly branched tubular reticulum. We determine the average
tubular diameter and find that it increases from 339 +/- 5 nm to
360 +/- 4 nm when changing from glucose to glycerol, that is, from
a fermentable to a nonfermentable carbon source. Moreover, this
change is associated with a 2.8-fold increase of the surface of the
reticulum, resulting in an average increase in volume of the
mitochondrial compartment by a factor of 3.0 +/- 0.2.
Best regards

Wolfgang Muss
Salzburg



----------
Von: phillipst-at-missouri.edu[SMTP:phillipst-at-missouri.edu]
Antwort an: phillipst-at-missouri.edu
Gesendet: Donnerstag, 02. Februar 2006 18:44
An: W.Muss-at-salk.at
Betreff: [Microscopy] Cytoviva system

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Does anyone have experience with the Cytoviva imaging system. It looks to
me (from their website at www.cytovita.com like it is a retrofit
condenser. They claim a resolution better than 50 nm. In fact, their
website says "The typical optical microscope provides a maximum theoretical
resolution limit of 240nm. With resolution below 100nm and detection below
50nm, CytoViva allows you to see details never before possible with
traditional microscopy. " Can someone explain to me how one can improve on
the maximum theoretical resolution?

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



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From: larry-at-cymru.freewire.co.uk
Date: Thu, 2 Feb 2006 14:51:23 -0600
Subject: [Microscopy] Re: ESEM chamber gas choices

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Hi Alwyn,

My information on this is not definite but, my understanding is that
different EDS manufacturers use different process for producing their
detector windows. Consequently, there are different consequences
between detectors depending on the gases used in the chamber. I can't
see that there would be a difference between Si(Li) and SDD but I can
understand a difference between manufacturers.

Best regards,
--
Larry Stoter
JEOL (UK) Ltd
tel: +44-(0)1707-377117, fax: +44-(0)1707-373254, e-mail: larrys-at-jeoleuro.com

PLEASE NOTE
1. Any mail other than plain text will be automatically deleted.
2. Any mail, legitimate or not, apparently or actually from hotmail,
netscape, yahoo or excite will automatically be deleted.
3. Mail with no subject or without a clear subject will be ignored :-)

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From: michael-at-shaffer.net
Date: Thu, 2 Feb 2006 16:31:55 -0600
Subject: [Microscopy] RE: Re: ESEM chamber gas choices

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Larry writes ...

} My information on this is not definite but, my understanding is that
} different EDS manufacturers use different process for producing their
} detector windows. Consequently, there are different consequences
} between detectors depending on the gases used in the chamber. I can't
} see that there would be a difference between Si(Li) and SDD but I can
} understand a difference between manufacturers.

To bring this back to SDD, my understanding is that all SDD detector chips
are manufactured by KETEK ... While individual SDD detector manufacturers
will choose to employ different windows. I am still waiting to find out the
source of information regarding a general query about SDDs' incompatibility
with water vapor.

Genuinely, Michael Shaffer :o)

SEM/MLA Laboratory Coordinator
http://www.mun.ca/creait/maf/
Inco Innovation Centre
Memorial University
St. John's, Newfoundland



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From: Judith_A_Ruiz-at-whirlpool.com
Date: Thu, 2 Feb 2006 19:41:52 -0600
Subject: [Microscopy] viaWWW: lab service for hire, How much do you charge

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Email: Judith_A_Ruiz-at-whirlpool.com
Name: Judith Ruiz

Organization: Whirlpool corporation

Title-Subject: [Filtered] How much do you charge

Question: For those who do lab service for hire, how much is the
going rate for SEM/EDX work? I only do work within our corporation
but we're putting together a cost of what our work would cost if it
was done outside.

Any help would be appreciated.

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From: mmstuason-at-yahoo.ca
Date: Thu, 2 Feb 2006 19:42:16 -0600
Subject: [Microscopy] viaWWW: type of CO2 do you use for CPD

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Email: mmstuason-at-yahoo.ca
Name: Lizette Tuason

Title-Subject: [Filtered] Type of CO2 for CPD

Question: Hi everyone,

What type of CO2 do you use for CPD? The CO2 we
currently have hooked up to our Denton DCP-1 is
supercritical fluid extraction grade but it costs
more than CAD$750. I looked up some online sites
where people mentioned that they use just
standard CO2 (in the US). Iķm not sure what
standard CO2 is and what the equivalent would be
here. Iķm buying from Praxair (Canada) and there
are several categories that Iķm not sure which
one I should be choosing.

Could anyone whoķs doing CPD tell me what CO2
you buy ń company and catalog number?

Thanks.

Lizette Tuason

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From: gary-at-gaugler.com
Date: Thu, 2 Feb 2006 20:20:13 -0600
Subject: [Microscopy] Re: viaWWW: lab service for hire, How much do you

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$200/hour....$150/hour.... $125/hour...$100/hour....pick a number.

This is a nice set of numbers but there is more to outsourcing than just
the hourly rate...IMO. If a specimen is prepared in-house and then
sent out-house, what is the out-house person to look for? Well, you
will have to spend x hours writing a detailed procedural instruction
for what you want to be analyzed. If the specimen deviates from this,
then what? What is your writing time worth? Nothing?

The point is that there is great value in having the requester sitting
by the SEM operator. Look here, look there, what is this, what is that?
If you outsource, you will get exactly or less than what you ask for
since the operator does not know what to do outside of your written
request. If the job is so mundane (how many are?) then this is not
a problem. The SEM is a very valuable tool for many areas of interest.
But given a 12mm diameter specimen stub, which 2u are of the most interest?
"I'll know it when I see it." Right. but they threw the specimen over
the wall to the SEM folks. The results thrown back may not match up
with what was needed. The variability of specimens means that there is
no simple approach to evaluation. Plus, the requester is not at the
SEM to know that they saw what they needed the first time.

gary g.

Disclaimer: I do not do this sort of ambiguous work. However, I do
perform SEM analysis of unknown specimens and known specimens but I
know what to look for.


At 05:44 PM 2/2/2006, you wrote:



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From: bfoster-at-mme1.com
Date: Fri, 3 Feb 2006 03:05:42 -0600
Subject: [Microscopy] Re: Cytoviva system

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Dear Tom,

You are partially right about CytoViva: it is a retrofittable condenser fitted with special optics and an illuminator system.

Re: Resolution:
I worked with Aetos at some length on the launch of CytoViva and have seen better than 90nm resolution (as measured with a Richardson test slide).

Re: how it works
Dr. Vitaly Vodyanoy, the inventor currently has a paper in progress which will explain the physics. My personal observations lead me to believe that there is some sort of resonance effect rather than the traditional scattering or diffraction we normally use for imaging. The result looks a lot like fluorescence (bright object/dark background), without the need for staining. It also has an interesting ability to optically section (much like DIC or confocal). We were able to watch spirochetes actually invading cells as well as see more detail in bacteria and some special marine cells.

I encourage you to visit their website, if only to see some interesting images. Their technical applications specialist, Dr. Tom Hasling, is usually quite happy to run samples and Byron Cheatham, their sales manager, can probably arrange for a demo in your lab, if you are serious about potential purchase.


The system compares extremely well to other systems on the market in the $150K range, at about 10% the cost. ... and there's no cost for looking!

Hope this was helpful,
Best regards,
Barbara Foster

Microscopy/Microscopy Education
313 S Jupiter Rd, Suite 100
Allen, TX 75002
P: 972-954-8011
W: www.MicroscopyEducation.com

P. S.
Need a good general reference or light microscopy text for the Spring semester? Call us today to learn more about "Optimizing LIght Microscopy". Copies still available through MME... even for class-room lots ... and we give quantity discounts. Call Ken Piel at (972)954-8011.







At 11:42 AM 2/2/2006, phillipst-at-missouri.edu wrote:



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From: bfoster-at-mme1.com
Date: Fri, 3 Feb 2006 03:15:06 -0600
Subject: [Microscopy] Re: viaWWW: LM, Anyone with "Digital Microscope"

Contents Retrieved from Microscopy Listserver Archives
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Hi, Brad

I'd recommend you also look into the Hi-Scope from Hirox-USA. They provide some really interesting imaging modes and have been well accepted by a number of major customers here in the US.

Caveat: MME has no financial interest in this product.

Hope this helpful,

Best regards,
Barbara Foster

Microscopy/Microscopy Education
313 S Jupiter Rd, Suite 100
Allen, TX 75002
P: 972-954-8011
W: www.MicroscopyEducation.com

P. S.
Need a good general reference or light microscopy text for the Spring semester? Call us today to learn more about "Optimizing LIght Microscopy". Copies still available through MME... even for class-room lots ... and we give quantity discounts. Call Ken Piel at (972)954-8011.



At 08:32 PM 1/30/2006, brad.huggins-at-bp.com wrote:



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From: M_Jarnik-at-fccc.edu
Date: Fri, 3 Feb 2006 08:51:46 -0600
Subject: [Microscopy] Re: viaWWW: type of CO2 do you use for CPD

Contents Retrieved from Microscopy Listserver Archives
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Lizette,

You don't need the really pure stuff. We use food-grade, and in one trial found it cleaner (less water, oil, and particulates) than the expensive, "pure" siphon CO2.
Just be sure to use a siphon-CO2 (comes in a cylinder with a siphon tube, so you get liquid, not gas, coming out), and spend the money for filters and dehydrating sieves.

Phil


We have the same Denton you do and use something what Airgas calls "bone
dry", cat. No. CDBD200S with siphon tube. In addition, we use the
Tousimis liquid CO2 filter (cat. No. 8784, I believe it helps). We pay
about US$ 65 for a cylinder, if I remember well. This seems to be giving
good results for CPD of biological material (mostly mouse embryos) for SEM.

Hope this helps,

Michal

mmstuason-at-yahoo.ca wrote:

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From: Jones_e1269-at-yahoo.com
Date: Fri, 3 Feb 2006 15:44:52 -0600
Subject: [Microscopy] viaWWW: Immuno EM

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Email: Jones_e1269-at-yahoo.com
Name: Jason Jones

Organization: The University of Texas Health Science Center at San Antonio

Title-Subject: [Filtered] Immuno EM

Question:

I am looking for a facility to do immuno-EM on the yeast Candida
albicans for us. We're interested in intracellular localization of a
soluble secretory
protein, secreted aspartyl protease (Sap2p) in wild-type and a
secretory mutant strain we have generated.

We have polyclonal antibodies to Sap2p, which have worked quite well
in the published literature for immunoEM, and we have also 6X-His
tagged
this protein in our wild-type and mutant strains.

IWe do not have immunoEM available as a core facility here.

Thank you.

Sincerely,

Jason Jones

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From: df-at-donnforbes.com
Date: Fri, 3 Feb 2006 15:45:13 -0600
Subject: [Microscopy] viaWWW: SEM for BWA Detection

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Email: df-at-donnforbes.com
Name: Donn Forbes

Organization: Arasil, Inc.

Title-Subject: [Filtered] SEM for BWA Detection

Question: Why isn't SEM a standard technology for detecting and
identifying viruses and bacteria used as biological warfare agents?

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From: bcarrington-at-slfc.org
Date: Fri, 3 Feb 2006 15:48:45 -0600
Subject: [Microscopy] AskAMicroscopist: K-8 Grade Grammar School

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Below is the result of your feedback form (NJZFM-ultra-55). It was
submitted by (bcarrington-at-slfc.org) from
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Email: bcarrington-at-slfc.org
Name: Brenda Carrington

Organization: The Learning Center

Education: K-8 Grade Grammar School

Location: Chesterfeild MO (St. Louis area)

Question: We are haing a microscope day on Feb. 27 from 10:00-3:00.
I don't know how to contact a local person to assist us.
We have over 100 kids grades 2-9.
We will be studying microbes and microscopes and using the GEMS materials.



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From: dsoren-at-umich.edu
Date: Fri, 3 Feb 2006 15:48:58 -0600
Subject: [Microscopy] Cleaning TEM grids

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Dear listers,

We have inherited lots of copper TEM grids that are decades old.
Many of them have a dark patina on them, and, on those grids, we have
been losing our sections during alcohol-based uranyl acetate
staining. The sections remain when we use water-based uranyl
acetate, however. We have tried sonicating the grids in ethanol,
acetone, or chloroform, none of which has solved our problem.

Does anyone have any suggestions for cleaning them so that we will
not lose our sections during staining, or should we pitch them? We
really do prefer to stain with alcohol- based uranyl acetate, since
it provides a more intense staining.

As always, thanks for any suggestions you might have.

Dotty Sorenson

Dorothy Sorenson
Microscopy and Image-analysis Laboratory
Department of Cell and Developmental Biology
University Of Michigan Medical School
4643 Medical Science Building II
1301 Catherine
Ann Arbor, MI 48109-0616
(734)763-1170
FAX (734)763-1166


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From: GBurgess-at-exchange.hsc.mb.ca
Date: Fri, 3 Feb 2006 16:08:11 -0600
Subject: [Microscopy] RE: Cleaning TEM grids

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Hi Dotty,
I would try putting your grids into a drying oven for at least 5 minutes
after you cut your ultrathin sections. After they are totally dried down,
you can take them out. If you forget and leave your grids in the oven
overnight, it doesn't seem to hurt anything at all. The heat seems to bond
the sections to the grids very well, such that I've never had an experience
of sections coming off the grids, regardless of the state of the grids.

To clean my grids, I just dip them into concentrated sodium hydroxide for a
few seconds, and then rinse them by dipping them a few times in distilled
water just before I pick up the sections. It sort of etches the grids.

Garry Burgess
Charge Technologist - Electron Microscopy
Health Science Centre
Winnipeg, Canada



Dear listers,

We have inherited lots of copper TEM grids that are decades old.
Many of them have a dark patina on them, and, on those grids, we have
been losing our sections during alcohol-based uranyl acetate
staining. The sections remain when we use water-based uranyl
acetate, however. We have tried sonicating the grids in ethanol,
acetone, or chloroform, none of which has solved our problem.

Does anyone have any suggestions for cleaning them so that we will
not lose our sections during staining, or should we pitch them? We
really do prefer to stain with alcohol- based uranyl acetate, since
it provides a more intense staining.

As always, thanks for any suggestions you might have.

Dotty Sorenson

Dorothy Sorenson
Microscopy and Image-analysis Laboratory
Department of Cell and Developmental Biology
University Of Michigan Medical School
4643 Medical Science Building II
1301 Catherine
Ann Arbor, MI 48109-0616
(734)763-1170
FAX (734)763-1166


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From: gstrout-at-ou.edu
Date: Fri, 3 Feb 2006 17:33:44 -0600
Subject: [Microscopy] Re: Cleaning TEM grids

Contents Retrieved from Microscopy Listserver Archives
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We clean our grids in acid alcohol, a mixture of 3% HCL in 95% ethanol.
Cover the grids in a small shell vial with the acid alcohol, swirl
intermittantly for 30 to 40 seconds or so and rinse well with 95%
ethanol. If the grids have a very dark patina then clean for a longer
time. You'll know when they are done because they'll have a shiny,
bright copper finish.
We usually clean many grids at a time, but you could clean them one by
one by dipping the grid in the acid alcohol and then rinsing in a stream
of 95% ETOH just before you use them. This works better on grids that
have been cleaned, but have set around for a while, long enough to have
developed a small oxidized layer. Dark patinas like the grids you have
will probably need to be put into a vial and cleaned as described above.

dsoren-at-umich.edu wrote:

} ----------------------------------------------------------------------------
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--
--
==================================================================
Greg Strout
Electron Microscopist, University of Oklahoma
WWW Virtual Library for Microscopy:
http://www.ou.edu/research/electron/www-vl/
e-mail: gstrout-at-ou.edu
Opinions expressed herein are mine and not necessarily those of
the University of Oklahoma
==================================================================



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From: walck-at-southbaytech.com
Date: Fri, 3 Feb 2006 20:16:09 -0600
Subject: [Microscopy] Cleaning TEM grids

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

Try soaking a few grids in store grade ammonia for about 1/2 hr to 1 hr
to see if the patina is removed at all. Rinse well in distilled water
and see what happens.

-Scott

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

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

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


-----Original Message-----
X-from: dsoren-at-umich.edu [mailto:dsoren-at-umich.edu]
Sent: Friday, February 03, 2006 1:53 PM
To: Walck-at-SouthBayTech.com

Dear listers,

We have inherited lots of copper TEM grids that are decades old.
Many of them have a dark patina on them, and, on those grids, we have
been losing our sections during alcohol-based uranyl acetate
staining. The sections remain when we use water-based uranyl
acetate, however. We have tried sonicating the grids in ethanol,
acetone, or chloroform, none of which has solved our problem.

Does anyone have any suggestions for cleaning them so that we will
not lose our sections during staining, or should we pitch them? We
really do prefer to stain with alcohol- based uranyl acetate, since
it provides a more intense staining.

As always, thanks for any suggestions you might have.

Dotty Sorenson

Dorothy Sorenson
Microscopy and Image-analysis Laboratory
Department of Cell and Developmental Biology
University Of Michigan Medical School
4643 Medical Science Building II
1301 Catherine
Ann Arbor, MI 48109-0616
(734)763-1170
FAX (734)763-1166


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From: dianavd-at-eye.usyd.edu.au
Date: Sun, 5 Feb 2006 22:06:54 -0600
Subject: [Microscopy] Re: Cleaning TEM grids

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

I've always cleaned grids by quickly passing through the flame of an
alcohol burner. Quick and easy. Works on my old grids, though they
aren't decades old!


Diana van Driel
Dept Ophthalmology
Sydney University
GPO Box 4337
Sydney, NSW
AUSTRALIA 2001

Phone 61 2 93827278
Mobile 0423 151614
FAX 61 2 93827318
On 4 Feb 2006, at 8:50 AM, dsoren-at-umich.edu wrote:

}
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}
} Dear listers,
}
} We have inherited lots of copper TEM grids that are decades old.
} Many of them have a dark patina on them, and, on those grids, we have
} been losing our sections during alcohol-based uranyl acetate
} staining. The sections remain when we use water-based uranyl
} acetate, however. We have tried sonicating the grids in ethanol,
} acetone, or chloroform, none of which has solved our problem.
}
} Does anyone have any suggestions for cleaning them so that we will
} not lose our sections during staining, or should we pitch them? We
} really do prefer to stain with alcohol- based uranyl acetate, since
} it provides a more intense staining.
}
} As always, thanks for any suggestions you might have.
}
} Dotty Sorenson
}
} Dorothy Sorenson
} Microscopy and Image-analysis Laboratory
} Department of Cell and Developmental Biology
} University Of Michigan Medical School
} 4643 Medical Science Building II
} 1301 Catherine
} Ann Arbor, MI 48109-0616
} (734)763-1170
} FAX (734)763-1166
}
}
} ==============================Original
} Headers==============================
} 7, 17 -- From dsoren-at-umich.edu Fri Feb 3 15:48:58 2006
} 7, 17 -- Received: from pushingtin.mr.itd.umich.edu
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} 7, 17 -- From: Dorothy Sorenson {dsoren-at-umich.edu}
} 7, 17 -- Subject: Cleaning TEM grids
} 7, 17 -- Date: Fri, 3 Feb 2006 16:47:15 -0500
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6, 20 -- From dianavd-at-eye.usyd.edu.au Sun Feb 5 22:06:54 2006
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6, 20 -- Subject: Re: [Microscopy] Cleaning TEM grids
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From: benada-at-biomed.cas.cz
Date: Mon, 6 Feb 2006 03:33:26 -0600
Subject: [Microscopy] Re: Cleaning TEM grids

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

Dear Dotty,
We were in the same situation some years ago. We cleaned the grids in the
following way:
- put the grids into small beaker (25ml) filled with 5% - 10% hydrochloric
acid
- let the grids to clean for some minutes, gently shake several times (at
the end of cleaning the grids should be shiny gold)
- remove the cleaning hydrochloric acid solution and wash the grids
several times with distilled water
- remove the distilled water as much as possible from the beaker and
replace it with acetone and let stand for some time
- pour the acetone with the grids into clean glass Petri dish filled with
filter paper
- remove the filter paper with the grids and put it into another glass
Petri dish and let dry out the rest of acetone
- that's all

The troubles with losing the sections during the staining could be
overcome by making the grids sticky:
- put 10 ml of chloroform into 25 ml glass beaker
- cut about 5 cm of 3M Magic Scotch tape and wash it in the chloroform
- remove the rest of the tape from the beaker
- put the grids onto filter paper and drop the "sticky solution" on them
using Pasteur pipette
- let the grids dry and use them for collecting the sections

Best regards from Prague
Oldrich
----------------------------------------
Oldrich Benada
Institute of Microbiology
Acad. Sci. CR
Videnska 1083
CZ - 142 20 Prague 4
Czech Republic
---------------------------------------




}
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} ----------------------------------------------------------------------------
}
} Dear listers,
}
} We have inherited lots of copper TEM grids that are decades old.
} Many of them have a dark patina on them, and, on those grids, we have
} been losing our sections during alcohol-based uranyl acetate
} staining. The sections remain when we use water-based uranyl
} acetate, however. We have tried sonicating the grids in ethanol,
} acetone, or chloroform, none of which has solved our problem.
}
} Does anyone have any suggestions for cleaning them so that we will
} not lose our sections during staining, or should we pitch them? We
} really do prefer to stain with alcohol- based uranyl acetate, since
} it provides a more intense staining.
}
} As always, thanks for any suggestions you might have.
}
} Dotty Sorenson
}
} Dorothy Sorenson
} Microscopy and Image-analysis Laboratory
} Department of Cell and Developmental Biology
} University Of Michigan Medical School
} 4643 Medical Science Building II
} 1301 Catherine
} Ann Arbor, MI 48109-0616
} (734)763-1170
} FAX (734)763-1166
}
}
} ==============================Original
} Headers==============================
} 7, 17 -- From dsoren-at-umich.edu Fri Feb 3 15:48:58 2006
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9, 25 -- From benada-at-biomed.cas.cz Mon Feb 6 03:33:26 2006
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From: dsams-at-schaferlabs.com
Date: Mon, 6 Feb 2006 19:16:53 -0600
Subject: [Microscopy] Mass Spectrometry Scientist / Senior Engineer at Schafer Corporation

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

Overview: Schafer Corporation, Sunol, California, is seeking a skilled and
innovative individual to add to our mass spectrometry group. SVL performs
materials characterization and related analytical services on commercial and
government contracts. The activities of the group include chemical and
elemental analysis of materials on a production basis, maintenance of
several mass spectrometers and ancillary equipment, development of new or
improved MS analysis techniques, and quality assurance of analytical data.

Schafer is a technically strong firm with a reputation for quality and
integrity. Schafer's reputation is a direct result of our dedicated,
motivated, talented, and creative staff that is responsible for developing
our outstanding business relationships with our customers. Our technical
capabilities are vast and growing to provide innovations for the future.

The successful candidate will work as part of a team responsible for
processing and analyzing small samples using laboratory instrumentation,
primarily Thermal Ionization and/or Secondary Ion Mass Spectrometers.

Responsibilities: Duties include instrument operation and data
interpretation; filament construction and preparation; standards loading; as
well as high vacuum, high voltage, and cryogenic system maintenance. The
successful candidate should have or be able to develop skills to process
samples, maintain equipment, and assist in developing and optimizing
analytical techniques.

Qualifications: The ideal candidate will have technical training in an
appropriate physical sciences field and related experience in a scientific
laboratory, preferably operating mass spectrometers or other analytical
equipment. Ability to work independently as well as part of a team is
required. Good customer service focus and commitment to quality are
required. Experience in the use of a light microscope, and sufficient
physical dexterity to perform micromanipulation tasks is highly desired, as
is knowledge of high vacuum and cryogenic principles.
Schafer is looking for people that have established safe laboratory skills
and exceptional aptitude for details including accurate record keeping.
Experience in laboratory data management (word processing, spread sheets and
databases) is also desired. Experience with mechanical systems or machining
skills would be helpful.
Other qualifications include:
. AA degree plus 10 years experience or Bachelor's degree in physical
science or engineering plus two years of technical experience.
. Demonstrated ability to solve technical problems.
. Experience in data evaluation and quality control.
. Must be a US citizen with the ability to obtain government security
clearance.
Apply at:
http://www.schafercorp.com/Careers/open.htm
http://jobs-schafer.icims.com/schafer_jobs/jobs/candidate/job.jsp?jobid=1106
&mode=view


==============================Original Headers==============================
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6, 20 -- From: "David Sams" {dsams-at-schaferlabs.com}
6, 20 -- To: {Microscopy-at-microscopy.com}
6, 20 -- Subject: Mass Spectrometry Scientist / Senior Engineer at Schafer Corporation
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From: RANGETS-at-AOL.COM
Date: Mon, 6 Feb 2006 22:27:40 -0600
Subject: [Microscopy] viaWWW: NEED SCHEMATICS LEO 440

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This Question/Comment was submitted to the Microscopy Listserver
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Email: RANGETS-at-AOL.COM
Name: BEN GHAFFAARI

Organization: RANGE TECHNICAL

Title-Subject: [Filtered] NEED SCHEMATICS

Question: DOES ANYONE HAVE AN EXTRA SET OF "LEO" 440
SCHEMATICS, WE CAN BUY OR PURCHASE, WHEN WE TRY
TO CONTACT THE MFR, WE DO NOT GET A RESPONSE, SO
MAYBE WE ARE USING AN INCORRECT CONTACT. IF
SOMEONE KNOWS WHO WE CONTACT FOR "LEO" PARTS
ANY SUGGESTIONS WILL BE APPRECIATED

BEN

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From: mcmahojt-at-ccf.org
Date: Mon, 6 Feb 2006 22:31:10 -0600
Subject: [Microscopy] viaWWW: Spurr Resin

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Email: mcmahojt-at-ccf.org
Name: Jim McMahon

Organization: Cleveland Clinic Foundation

Title-Subject: [Filtered] Spurr Resin

Question: We have been having great difficulty with the newly
formulated Spurr Resin and in particular the ERL component. Not only
is Spurr no longer low viscosity but we find that its penetration and
polymerization to be far inferior to the original. We are prepared
to abandon the resin in favor of another such as PolyBed or Araldite.
But first I would like to know if anyone else has had similar
problems and was able to solve them.

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

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From: paul_hazelton-at-umanitoba.ca
Date: Mon, 6 Feb 2006 23:49:50 -0600
Subject: [Microscopy] Re: viaWWW: Spurr Resin

Contents Retrieved from Microscopy Listserver Archives
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jim

for what it is worth, i've recently re-tried a commercial version of
Epon 812 on a cell suspension. i found the cells would not settle
through the resin mix, and could not be pelleted. a parallel embedment
in the new Spurr with ERL behaved very well. in short, whatever is said
about low viscosity editions of Epon, neither they, nor the original
were ever thin enough to allow embedments of free cells and i personally
found them more than a little difficult to cut. but then i've always
found Epon, in whatever formulation, prone to static and difficult to
section.

on the other hand, the problem may really be an issue of density of the
medium, causing the the cells to be bouyant in the medium. does anyone
want to contribute knowledge on that?

having said that, i do find the new ERL component to be a little
brittle, but very sectionable.

paul



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From: M_Jarnik-at-fccc.edu
Date: Tue, 7 Feb 2006 08:35:31 -0600
Subject: [Microscopy] Poly-L-lysine coated coverslips

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

We would need some to improve cell attachment for a time course
experiment. Surprisingly, we were not able to locate any coverslips
(unlike slides). Does anybody have recommendations? (Of course, we can
always make our own.)

Thanks,

Michael




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From: mcauliff-at-umdnj.edu
Date: Tue, 7 Feb 2006 08:43:50 -0600
Subject: [Microscopy] Re: Spurr Resin

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

I have recently encountered problems with Spurr as well. I embed 100
micron thick Vibratome sections of brain so penetration should not be a
problem. I am using the same processing proceedure I have used for many,
many years on much larger tissue blocks. The center of the sections
seems to be poorly infiltrated and the block is very brittle. Cutting
intact 1 micron sections is nearly impossible. The viscosity seems
'normal', in other words, like I am used to with Spurr and I am not
familiar with any 'new' or 'improved' version of the mix or its
components. I will cut some more blocks and talk to my supplier (a very
reputable EM supplier I have used for 30 years) to see if I can shed
some light on this problem.

Geoff

mcmahojt-at-ccf.org wrote:

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--
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Geoff McAuliffe, Ph.D.
Neuroscience and Cell Biology
Robert Wood Johnson Medical School
675 Hoes Lane, Piscataway, NJ 08854
voice: (732)-235-4583; fax: -4029
mcauliff-at-umdnj.edu
**********************************************



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From: paul_hazelton-at-umanitoba.ca
Date: Tue, 7 Feb 2006 08:53:50 -0600
Subject: [Microscopy] Re: Poly-L-lysine coated coverslips

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

Michael

Depends on what you want to do. If it is for EM you can use the
Thermonox coverslips. I think all EM suppliers sell them, along with
the general suppliers.

However, if you want to do IF or confocal work the thermonox will quench
the reactions somewhat so you will have to do glass. I've used straight
glass, but many cell lines will not stick well to glass. Undoubtably
someone will provide the name of a supplier of pre-treated slips if they
are out there somewhere.

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: DRK-at-SHCC.org
Date: Tue, 7 Feb 2006 09:55:36 -0600
Subject: [Microscopy] viaWWW: Spurr Resin

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

Regarding the brittle nature of Spurrs resin due to the new ERL component,
we have solved our problems by combining the components according to the
"soft" recipe:

ERL 10g
DER 7g
NSA 26g
DMAE 0.4ml

This mixture results in blocks about as hard as the old "hard" formulation
using VCD. We have not noticed a lack of tissue penetration during
infiltration, but we may be less sensitive to this issue. Polymerization is
still at 70 deg C for 17 hours.

Best wishes,

Doug


Douglas R. Keene
Assistant Investigator
Micro-Imaging Center
Shriners Hospital for Children
3102 S.W. Sam Jackson Park Road
Portland, Oregon 97239
503-221-3434
drk-at-shcc.org

-----Original Message-----
X-from: mcmahojt-at-ccf.org [mailto:mcmahojt-at-ccf.org]
Sent: Monday, February 06, 2006 8:38 PM
To: drk-at-SHCC.org

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Email: mcmahojt-at-ccf.org
Name: Jim McMahon

Organization: Cleveland Clinic Foundation

Title-Subject: [Filtered] Spurr Resin

Question: We have been having great difficulty with the newly
formulated Spurr Resin and in particular the ERL component. Not only
is Spurr no longer low viscosity but we find that its penetration and
polymerization to be far inferior to the original. We are prepared
to abandon the resin in favor of another such as PolyBed or Araldite.
But first I would like to know if anyone else has had similar
problems and was able to solve them.

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

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==============================Original Headers==============================
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From: gary-at-gaugler.com
Date: Tue, 7 Feb 2006 10:06:45 -0600
Subject: [Microscopy] Re: viaWWW: NEED SCHEMATICS LEO 440

Contents Retrieved from Microscopy Listserver Archives
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I think that they are responding in their own way.
Their answer is "No."

gary g.


At 08:30 PM 2/6/2006, you wrote:



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From: scanning-at-fams.org
Date: Tue, 7 Feb 2006 12:54:34 -0600
Subject: [Microscopy] Scanning 2006 Abstract Deadline is February 18

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

Dear Group:

Kindly note the abstract submission deadline for SCANNING 2006 is
February 18. We hope to see you April 25-27 in D.C.!

Best regards,


Phaedra McGuinness
Managing Editor
SCANNING, The Journal of Scanning Microscopies
P.O. Box 485
Mahwah, NJ 07430
Tel: (201) 818-1010 * Fax: (201) 818-0086 *email: scanning-at-fams.org *
Web: www.scanning.org


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From: paul_hazelton-at-umanitoba.ca
Date: Tue, 7 Feb 2006 15:19:26 -0600
Subject: [Microscopy] viaWWW: Spurr Resin

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

doug

what you recommend in your note is the logical solution. actually, i
had meant to try the same change for my last embedment of monolayers to
correct for the hardness/brittleness. unfortunately, i got distracted
and forgot. i suspect that to get the medium hardness you may need 8-9
ml of the ERL component.

also, i used to use 0.4gm DMAE but changed that to .3gm to give a longer
pot life. also, higher catalyst concentration could have an effect on
viscosity and penetration.

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: WAHeeschen-at-dow.com
Date: Tue, 7 Feb 2006 16:05:53 -0600
Subject: [Microscopy] Job Posting from The Dow Chemical Company

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http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Job Posting from The Dow Chemical Company

An immediate opening exists for a skilled microscopist in Dow's Global
Analytical Sciences Laboratory part of Dow's Corporate R&D function. We are
seeking an individual with a passion for characterization science and a
desire to both apply this science to complex industrial problems and to
advance the technology. A Ph.D. in science is preferred, but an individual
with a Masters degree and relevant work experience will also be considered.
Working experience in electron microscopy is essential with skills in
scanning and transmission electron microscopy, focused ion beam methods,
energy dispersive x-ray analysis, electron energy loss spectroscopy,
electron diffraction and microtomy preferred. A background in catalysis is
a plus. Excellent written and oral communication skills (English) are
essential with an ability to work in a globally diverse team environment.
The position supports new product development activities working from a
laboratory with a FEG TEM-STEM, 2 TEMs, a FIBSEM, EPMA, FEGSEM with
cryotransfer system, a full complement of scanned probe and light
microscopes, SAXS, XPS and TOF-SIMS. The position is located in Midland,
Michigan. Applicants must have the ability to work in the USA.

Dow is a leader in science and technology, providing innovative chemical,
plastic and agricultural products and services to many essential consumer
markets. With annual sales of $40 billion, Dow serves customers in 175
countries and a wide range of markets that are vital to human progress:
food, transportation, health and medicine, personal and home care, and
building and construction, among others. Committed to the principles of
sustainable development, Dow and its 43,000 employees seek to balance
economic, environmental and social responsibilities. References to "Dow" or
the "Company" mean The Dow Chemical Company and its consolidated
subsidiaries unless otherwise expressly noted.

Please submit curriculum vitae and a list of references to Dr. John
Blackson, Building 1897, The Dow Chemical Company, Midland, MI 48667.

Best Regards,
Bill
William A. Heeschen, Ph.D.
Microscopy, Digital Imaging
The Dow Chemical Company
Midland, MI 48674
waheeschen-at-dow.com



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From: scanning-at-fams.org
Date: Wed, 8 Feb 2006 08:04:08 -0600
Subject: [Microscopy] Abstract deadline for Scanning 2006

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Dear Group:

Kindly note the abstract submission deadline for SCANNING 2006 is
February 18. We hope to see you April 25-27 in D.C.!

Best regards,


Phaedra McGuinness
Managing Editor
SCANNING, The Journal of Scanning Microscopies
P.O. Box 485
Mahwah, NJ 07430
Tel: (201) 818-1010 * Fax: (201) 818-0086 *email: scanning-at-fams.org *
Web: www.scanning.org


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From: mcauliff-at-umdnj.edu
Date: Wed, 8 Feb 2006 15:25:23 -0600
Subject: [Microscopy] Spurr's resin update

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

Greetings all:

I just got off the phone with Stacy (the owner) at Electron
Microscopy Sciences. I discussed my recent problems with Spurr's resin
with her. She told me that one component, ERL 4206, is no longer
maunfactured due to its high toxicity. It has been replaced with ERL
4221. She thinks that this is the cause of recent problems with Spurr
embedments. She has discussed this with others (I did not ask who) and
the consensus recommendations are to prolong dehydration in graded
ethanols to 20 minutes each step, make the 1:1 prop. oxide:Spurr step 4
hours and make the first change of pure resin 6 hours or longer. Since
longer times in solvents are known to extract cytoplasmic components
(and I cannot imaging that a 100 micron Vibratome section of CNS needs
20 min. per change of graded ethanol) I am going back to Epon
substitutes. Also, my tissues are in a second change of fresh resin on a
rotator overnight and then spend at least 1 hour under vacuum, I can't
see how infiltration could be a problem. I have used both the soft and
firm mixtures with different tissues and had inconsistant block textures
and infiltration problems with both. I don't have these problems with an
Araldite kit I bought at the same time. I can't risk knock-out mice in
long-term treatment/recovery experiments to reagents I cannot be sure of.

Geoff

--
**********************************************
Geoff McAuliffe, Ph.D.
Neuroscience and Cell Biology
Robert Wood Johnson Medical School
675 Hoes Lane, Piscataway, NJ 08854
voice: (732)-235-4583; fax: -4029
mcauliff-at-umdnj.edu
**********************************************



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From: Daniel.Salamon-at-nrc-cnrc.gc.ca
Date: Wed, 8 Feb 2006 15:33:09 -0600
Subject: [Microscopy] S4800 STEM scintillator size

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

Hi everyone,

I will be experimenting with alternative scintillators for our Hitachi
S4800 STEM unit. As I was advised to no touch the scintillator in the
microscope, I hope someone will be able to offer measurements for the
original S4800 scintillator.

Thanks,
Daniel Salamon
Technical Officer, Electron Microscopy

National Institute for Nanotechnology, NRC
W6-017A ECERF Bldg, 9107-116 Street
Edmonton, AB. T6G 2V4

Phone: Office (780) 492 8878
Lab (780) 492 8872
DocuFax: (780) 492 8632



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From: gerd.leitinger-at-meduni-graz.at
Date: Thu, 9 Feb 2006 06:45:00 -0600
Subject: [Microscopy] EM: precautions when en bloc staining

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

Dear list,

We are just starting an experiment with en-bloc staining of tissue in uranyl
acetate in 70% ethanol.
It seems to me that since the blocks are larger than thin sections, they
probably take up more radioactive material- so should we take any
precautions when handling the blocks after the resin has cured? (i.e. store
the blocks in separate special containers, wear gloves when sectioning or
are there any special cleaning procedures for the knife?).

thank you for sharing your knowledge

Gerd

Dr. Gerd Leitinger

Institut für Zellbiologie, Histologie und Embryologie
Medizinische Universität Graz
Harrachgasse 21
A-8010 Graz
Austria

Tel. ++43 316 380 4237
Fax. ++43 316 380 9625
Mailto: Gerd.Leitinger-at-meduni-graz.at



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From: donc-at-asmicro.com
Date: Thu, 9 Feb 2006 06:56:12 -0600
Subject: [Microscopy] AFM equipment wanted

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

I am seeking to buy NanoScope AFM equipment, such as MultiMode or Dimension,
to use with my Nanoscope IIIA controller. If you have anything to sell,
please contact me offline.

regards,
Don Chernoff
==================================
Advanced Surface Microscopy, Inc. E-Mail: donc-at-asmicro.com
3250 N. Post Rd., Ste. 120 Voice: 317-895-5630
INDIANAPOLIS IN 46226 USA Toll free: 800-374-8557 (in USA & Canada)
web: http://www.asmicro.com Fax: 317-895-5652
[business activities: analytical services in AFM, AFM probes, consulting,
training,
calibration and test specimens, calibration and measurement software,
used NanoScope equipment.]


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From: cumings-at-umd.edu
Date: Thu, 9 Feb 2006 08:35:58 -0600
Subject: [Microscopy] Wanted: Gatan 622SC

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

Hello microscopists,

We are looking to purchase a second-hand Gatan 622SC camera
(200kV-compatible) in decent working condition to install on our JEOL
2100 TEM. We are hoping to find one with a working intensifier and an
intact scintillator, although any problems are possibly acceptable.
We would be able to pay for it using a University of Maryland PO. The
amount of money might allow you to upgrade your TEM to a digital CCD
camera from AMT.

Please do not reply to the listserver. Direct any inquiries or offers
directly to me at the email address below.

Many thanks,

-John

--
John Cumings
cumings-at-umd.edu
Assistant Professor
Department of Materials Science and Engineering
University of Maryland
College Park, MD 20742-2115

office (301) 405-0789 (1246 Kim Building)
fax (301) 314-8164
--


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From: hyi-at-emory.edu
Date: Thu, 9 Feb 2006 10:06:53 -0600
Subject: [Microscopy] Bacterial Pili

Contents Retrieved from Microscopy Listserver Archives
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Dear Microscopists:


We have a few users study bacterial pili using negative staining with
PTA (pH 6.5). We have got some pretty nice images. But often time, we
could not find pili even on the positive control.  When we did see
pili, they were not on all bacteria in the same sample. Is this a
common phenomenon? Do bacteria lose their pili easily when the external
condition is not favorable? if that is the case, what should be done to
minimize the loss.

Thank you in advance. 

Hong
Emory EM


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From: TindallR-at-missouri.edu
Date: Thu, 9 Feb 2006 10:24:23 -0600
Subject: [Microscopy] Bacterial Pili

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Hong,

We often have this same problem. At least in our lab, flagella and pili seem to hide when we're trying to find them and show up when we don't care if we see them or not.

As a rule, they seem to be easily detached by rough handling, including centrifuging, and maybe by the staining process itself, since we often find them isolated from the bacteria on the grid.

One group of researchers gets around this problem by growing the bacteria on carbon coated grids and staining them in situ. Check JOURNAL OF BACTERIOLOGY, Feb. 2005, p. 1173-1181 for images. For preparation details this article references Brown et al, 2001. Immunocytochemical localization of HrpA and HrpZ supports a role for the Hrp pilus in the transfer of effector proteins from Pseudomonassyringae pv. tomato across the host plant cell wall. Mol. Plant-Microbe Interact. 14:394-404. I have not yet read the last article, so I can't comment on it.

Good luck.

Randy

Randy Tindall
EM Specialist
Electron Microscopy Core Facility---We Do Small Well!
W122 Veterinary Medicine
University of Missouri
Columbia, MO 65211
Tel: (573) 882-8304
Fax: (573) 884-2227
Email: tindallr-at-missouri.edu
Web: http://www.emc.missouri.edu







-----Original Message-----
X-from: hyi-at-emory.edu [mailto:hyi-at-emory.edu]
Sent: Thursday, February 09, 2006 10:08 AM
To: Tindall, Randy D.

Dear Microscopists:


We have a few users study bacterial pili using negative staining with PTA (pH 6.5). We have got some pretty nice images. But often time, we could not find pili even on the positive control.  When we did see pili, they were not on all bacteria in the same sample. Is this a common phenomenon? Do bacteria lose their pili easily when the external condition is not favorable? if that is the case, what should be done to minimize the loss.

Thank you in advance. 

Hong
Emory EM


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From: Edward.Calomeni-at-osumc.edu
Date: Thu, 9 Feb 2006 11:18:04 -0600
Subject: [Microscopy] Bacterial Pili

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Hong,

Rough handling may contribute to loss of pili, however the PTA itself is
probably the culprit. Try changing the pH either up or down. Way back when,
I did a study with rotavirus and various negative stains. Bottom line is
that the length of time of staining, pH and concentration all contributed to
the destruction (preservation) of the viral particles. If I remember
correctly, the length of time was the most important factor. Also try using
either uranyl acetate or ammonium molybdate as the negative stain.

Best of luck,

Ed


Edward P. Calomeni
Director EM Lab - Pathology
The Ohio State University
M018 Starling Loving Hall
320 W. 10th Ave.
Columbus, OH 43210
614-293-5580 (office)
614-293-8806 (lab)
edward.calomeni-at-osumc.edu
-----Original Message-----
X-from: TindallR-at-missouri.edu [mailto:TindallR-at-missouri.edu]
Sent: Thursday, February 09, 2006 11:29 AM
To: Edward Calomeni

Hong,

We often have this same problem. At least in our lab, flagella and pili seem
to hide when we're trying to find them and show up when we don't care if we
see them or not.

As a rule, they seem to be easily detached by rough handling, including
centrifuging, and maybe by the staining process itself, since we often find
them isolated from the bacteria on the grid.

One group of researchers gets around this problem by growing the bacteria on
carbon coated grids and staining them in situ. Check JOURNAL OF
BACTERIOLOGY, Feb. 2005, p. 1173-1181 for images. For preparation details
this article references Brown et al, 2001. Immunocytochemical localization
of HrpA and HrpZ supports a role for the Hrp pilus in the transfer of
effector proteins from Pseudomonassyringae pv. tomato across the host plant
cell wall. Mol. Plant-Microbe Interact. 14:394-404. I have not yet read the
last article, so I can't comment on it.

Good luck.

Randy

Randy Tindall
EM Specialist
Electron Microscopy Core Facility---We Do Small Well!
W122 Veterinary Medicine
University of Missouri
Columbia, MO 65211
Tel: (573) 882-8304
Fax: (573) 884-2227
Email: tindallr-at-missouri.edu
Web: http://www.emc.missouri.edu







-----Original Message-----
X-from: hyi-at-emory.edu [mailto:hyi-at-emory.edu]
Sent: Thursday, February 09, 2006 10:08 AM
To: Tindall, Randy D.

Dear Microscopists:


We have a few users study bacterial pili using negative staining with
PTA (pH 6.5). We have got some pretty nice images. But often time, we could
not find pili even on the positive control.  When we did see pili, they were
not on all bacteria in the same sample. Is this a common phenomenon? Do
bacteria lose their pili easily when the external condition is not
favorable? if that is the case, what should be done to minimize the loss.

Thank you in advance. 

Hong
Emory EM


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From: tivol-at-caltech.edu
Date: Thu, 9 Feb 2006 12:48:20 -0600
Subject: [Microscopy] Re: EM: precautions when en bloc staining

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On Feb 9, 2006, at 4:45 AM, gerd.leitinger-at-meduni-graz.at wrote:

} We are just starting an experiment with en-bloc staining of tissue in
} uranyl
} acetate in 70% ethanol.
} It seems to me that since the blocks are larger than thin sections,
} they
} probably take up more radioactive material- so should we take any
} precautions when handling the blocks after the resin has cured? (i.e.
} store
} the blocks in separate special containers, wear gloves when sectioning
} or
} are there any special cleaning procedures for the knife?).
}
} thank you for sharing your knowledge
}
Dear Gerd,
There is still a very small amount of radioactive material in the
block; furthermore, uranium has a very long half-life, thus very small
activity, and the alpha particles it emits have a short range, so any
decays except those nearer the surface of the block than the thickness
of the dead layer of your skin will not result in much radiation
leaving the block. You can easily measure this with an ionization
chamber (to detect the x- and gamma-radiation that make up most of the
escaping radiation). That said, the laws regarding the handling of
radioactive materials--even those with very little activity--vary from
country to country, locale to locale, and sometimes for different
institutions within a particular locale, so your safety office may
mandate special procedures for storage and handling.
Yours,
Bill Tivol, PhD
EM Scientist and Manager
Cryo-Electron 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: nairvinods-at-gmail.com
Date: Thu, 9 Feb 2006 13:08:22 -0600
Subject: [Microscopy] Re: Bacterial Pili

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Dear Hong,
I have been using 2% Uranyl acetate to visualize bacterial pili.

I have been using 2% Uranyl acetate to visualize pili. I have had to
play with the time of staining and rinsing in order to get a good
contrast to visualize the pili on different bacterial genera. I have
noticed that not all bacteria show the same distribution of pili.
I have also noticed that the way you grow the bacteria also affect
visualization of pili. For our bacteria I have noticed that I get far
better results when I grow them up in stationary cultures without
shaking them.

What I have been seeing it the bacteria behave differently when in
aggregation versus solitude. This is more so owing to the several
differnt kinds of pili each bacterial strain is capable of forming. If
the bacteria you are working with produces different types of pili
then you will see differences between the bacterial cells visualized
on the same grid.

I have had the same strain of bacteria producing really small pili
which were visualized only at a very high mag.
Hope that was helpful
regards,
Vinod
Graduate Student
Dept Of Biology
New Mexico State University
On 2/9/06, hyi-at-emory.edu {hyi-at-emory.edu} wrote:
}
}
}
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} Dear Microscopists:
}
}
} We have a few users study bacterial pili using negative staining with
} PTA (pH 6.5). We have got some pretty nice images. But often time, we
} could not find pili even on the positive control. When we did see
} pili, they were not on all bacteria in the same sample. Is this a
} common phenomenon? Do bacteria lose their pili easily when the external
} condition is not favorable? if that is the case, what should be done to
} minimize the loss.
}
} Thank you in advance.
}
} Hong
} Emory EM
}
}
} ==============================Original Headers==============================
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From: icmicroanalysis-at-cox.net
Date: Thu, 9 Feb 2006 13:09:33 -0600
Subject: [Microscopy] Stains/etches for delineating source/drains diffusions on semiconductor die cross sections

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


I'm compiling a list of chemical stains/etches for delineating N+ and P+
source/drain implants/diffusions on semiconductor die cross sections. Any
suggestions?


==============================Original Headers==============================
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3, 22 -- From: "IC Microanalysis" {icmicroanalysis-at-cox.net}
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3, 22 -- Subject: Stains/etches for delineating source/drains diffusions on semiconductor die cross sections
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From: hagglundk1-at-nku.edu
Date: Fri, 10 Feb 2006 10:11:00 -0600
Subject: [Microscopy] Follow up ESEM gas choices

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

I put together a summary of the responses to my query on ESEM gas
choices. People responded that they had experimented with Nitrogen,
Nitrous Oxide, Oxygen, and Helium. Opinions were mixed on the
performance of various gases (see below), but in general I feel
comfortable trying Nitrogen in the near future. I received warnings
that Helium can damage photomultipliers and Argon can damage ion pumps.
Because of safety issues, we will most likely avoid oxygen and nitrous
oxide for the time being.

FEI also contacted me and reminded that before experimenting with
various gases in the ESEM, I should check with them to make sure that
warranties are not voided by using any particular gas. If a gas could
damage your system or components, the manufacturer should be able to
tell you before you damage your system. Always check with the
manufacturer before changing major system parameters.

My detector issue drew a number of separate questions and queries, and
we are working separately with the manufacturers on this. We did learn
that we can collect spectra with the SDD crystal at room temperature.
This eliminates the potential for vapor condensation on the crystal
surface. There is some peak broadening and a slightly higher background
when we do this, but it does provide us with bulk data and may prove to
be our final solution.

I also learned that many bulk gases also contain amounts of moisture
that might condense under variable pressure conditions. Ultra high
purity, low moisture gas, is sometimes considerably more expensive than
bulk lab gas. I have to check with my vendor to find out what kind of
moisture my tank might contain before hooking it to the ESEM.

Thanks again for everyone's help on this. The list has proven itself
invaluable again.

Karl

-----
"I have used water vapor, N2 and Argon. None of them gave as good a
signal (image) as the water vapor did.... The Ionization of the other
gasses is apparently not that good and give no nice images"
-----
"I have been using dry nitrogen with my variable pressure instrument. We
vent the system with the same gas. My EDS system seems to work well but
have not run extensive tests to determine what effects the gas has other
than the beam scattering issues."
-----
" A disadvantage to using helium is your PMT's could be damaged, if it
is released into the room. Also, another gas you could be careful with
is argon, as it is a ion-pump poison (if released into a ion-pumped
system).
Nitrogen sounds friendly to me"
-----
"In my experience, if you can't use oxygen the next best gas to use is
argon - easily obtainable and provides good imaging conditions. Air and
nitrogen should be avoided, since the nitrogen breaks down at too low a
voltage and you can't get much signal. Some European groups looked at a
wider range of imaging gases and nitrous oxide I believe did very well,
although I have not tried it. I have tried oxygen and it works well, but
tends to ruin the pump oil fairly quickly."
-----
"The Quanta 200 uses a W filament. Unless it is a SFEG. If there are
no ion pumps, you should be able to use Nitrogen or He. If there is an
ion pump, do not use He. It will kill the pump.

I found that for VP (20-120Pa) that N2 works fine and is better than air
(less vacuum issues). So, for ESEM, I would think that N2 ought to be
the gas of choice as well."
-----
" We routinely use He in our Hitachi VP-SEM. It has a tungsten gun and
no ion pumps, so there is no problem with fouling those pumps as Gary
Gaugler mentioned.

If you consider that He is a monoatomic species with a weight of 4 and
that nitrogen is diatomic with a weight of 28, your gut can tell you
that He will scatter less than N2 or room air. That is our experience
and we can see the effect in iamges.

There is no effect on the x-ray signals from He that we have ever seen.
He x-rays are below our low-energy discriminator. I suppose one could
see N or O x-rays due to the gas, but I suppose their contribution is
much less than from the sample. You could arrange a test to evaluate
that, say you collected spectra from a beryllium or boron sample using
high vacuum, helium, and nitrogen."
-----
Original Post:
---
----

I am interested in hearing what type of gases people are using in their
ESEM applications. We recently learned our silicon drift detector is
incompatible with water vapor, and we apparently cannot complete any
x-ray microanalysis while working in standard ESEM or VP modes (I was
pretty surprised to learn this). We are thinking that using a dry gas
such as nitrogen or helium will allow us to work in ESEM modes and use
our x-ray system as well, as there is no potential for moisture
condensing on the crystal surface.

Our ESEM is a Quanta 200, and it is not equipped with the peltier stage,
so we mainly use the environmental modes to alleviate charging in
uncoated samples. The x-ray system was purchased with the microscope
about four years ago. I would rather not discuss the name of the x-ray
vendor on the list, as we have a significant investment in this detector
and do not foresee finding the money to purchase a new one soon.

Our first concern is the impact of the gas on spectra. We can coat
samples and run them in high vacuum mode, but customers like spectra
that represent the sample and not the coating material. We periodically
have samples that cannot be coated, so ESEM becomes critical for our
imaging needs. If the gas is going to have a dramatic impact on signal,
are we better off coating and running high vacuum?

Is there a best choice for chamber gas? Our service engineer recommends
nitrogen as having benefits for keeping the system clean. We can set it
up fairly quickly, and I have a spare tank and regulator. I have heard
of people using helium and believe that I read somewhere that there was
some advantage to using helium.

I am still trying to get information from the vendor on whether this
will allow us to use the x-ray and ESEM simultaneously, or whether we
have options with different collimators or windows. Their responses
have been pretty slow coming.
_____

Karl Hagglund
Biological Sciences, SC-102B
Northern Kentucky University, Nunn Drive
Highland Heights, KY 41099
859-572-5238
http://semlab.nku.edu



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From: kenconverse-at-qualityimages.biz
Date: Fri, 10 Feb 2006 10:20:25 -0600
Subject: [Microscopy] S4800 STEM scintillator size

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

Daniel,
If you contact Gene Taylor at M. E. Taylor Engineering

Phone 301-774-6246

Fax 301-774-6711

www.semsupplies.com

I'm sure he can give you what you need. Scintillators have been his
specialty for decades. Also, many of the other EM supply companies should
be able to provide what you're looking for.

Disclaimer: A number of years ago when my business was larger, I was a
distributor of Taylor supplies and have many happy customers.

Ken Converse
Owner

QUALITY IMAGES
Servicing Scanning Electron Microscopes
Since 1981
474 So. Bridgton Rd.
Bridgton, ME 04009
207-647-4348
Fax 207-647-2688
kenconverse-at-qualityimages.biz
qualityimages.biz


-----Original Message-----
X-from: Daniel.Salamon-at-nrc-cnrc.gc.ca [mailto:Daniel.Salamon-at-nrc-cnrc.gc.ca]
Sent: Wednesday, February 08, 2006 4:35 PM
To: kenconverse-at-qualityimages.biz

Hi everyone,

I will be experimenting with alternative scintillators for our Hitachi
S4800 STEM unit. As I was advised to no touch the scintillator in the
microscope, I hope someone will be able to offer measurements for the
original S4800 scintillator.

Thanks,
Daniel Salamon
Technical Officer, Electron Microscopy

National Institute for Nanotechnology, NRC
W6-017A ECERF Bldg, 9107-116 Street
Edmonton, AB. T6G 2V4

Phone: Office (780) 492 8878
Lab (780) 492 8872
DocuFax: (780) 492 8632



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==============================Original Headers==============================
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From: RossLM-at-missouri.edu
Date: Fri, 10 Feb 2006 11:55:40 -0600
Subject: [Microscopy] 2 contract positions at Monsanto in St. Louis

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

I am posting the following ad for Dr. Jingyue Liu at Monsanto. Please
contact Dr. Liu for further information, email address at the bottom
of the request.

Lou Ross


Two Contract Researchers Are Needed in the Catalyst Characterization
Group of Monsanto Company

Job Location: Creve Coeur Campus, St. Louis, Missouri 63167

The following two openings (one year contract researchers) are
immediately available:

1) Research associate for TEM sample preparation. Required skills:
extensive experience and expertise in ultramicrotoming thin sections
of catalyst powders or other nanophase materials for transmission
electron microscopy observation. This job requires strong hands-on
skills and new method development for unique samples. Experience
with sample preparation equipments such as high vacuum carbon
coating, embedding, fixing and staining biological tissues, and
maintenance of sample preparation facility is highly desirable.
Experience in operating SEM instruments is a plus.

2) Research associate for catalyst preparation, treatment and
characterization. Required skills: extensive experience in preparing
model and practical catalysts or nanoparticles and TEM
characterization of such samples. Experience in catalyst treatment
and testing is a plus. Strong hands-on skills and demonstrated
capability of designing complex experiments are required for this job.

Both jobs require extensive hands-on experiences in research labs.
The following competencies are required: innovation in solving
challenging problems; good communication and interpersonal skills;
teamwork skills and results orientation.

Since Monsanto does not directly hire contract researchers, the
selected candidates will work for a contract agency. Interested
parties please send your resume and application letter to:
Jingyue.liu-at-monsanto.com.

--
Senior Electron Microscope Specialist
Electron Microscopy Core Facility
W136 Veterinary Medicine
University of Missouri
Columbia, MO 65211-5120
(573) 882-4777, fax 884=2227
email: rosslm-at-missouri.edu
http://www.emc.missouri.edu

==============================Original Headers==============================
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From: ldmm-at-risc4.numis.northwestern.edu
Date: Fri, 10 Feb 2006 12:25:19 -0600
Subject: [Microscopy] Texts that combine TEM + scanning probe techniques

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

I am comtemplating broadening a mainly TEM undergrad course (with a lab)
to include some scanning probe techniques. Does anyone know of reasonable
texts which have some coverage?

-----------------------------------------------
Laurence Marks
Department of Materials Science and Engineering
MSE Rm 2036 Cook Hall
2220 N Campus Drive
Northwestern University
Evanston, IL 60201, USA
Tel: (847) 491-3996 Fax: (847) 491-7820
email: L - marks -at- northwestern . edu
http://www.numis.northwestern.edu
-----------------------------------------------



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From: anna8261-at-yahoo.com
Date: Sat, 11 Feb 2006 17:21:03 -0600
Subject: [Microscopy] AskAMicroscopist: heterostructure InP/InGaAsP

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

Below is the result of your feedback form (NJZFM-ultra-55). It was
submitted by (anna8261-at-yahoo.com) from
http://www.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html
on Friday, February 10, 2006 at 02:52:59
---------------------------------------------------------------------------

Email: anna8261-at-yahoo.com
Name: anna naghshegar

Organization: pooysh

Education: Graduate College

Location: tehran, iran

Question: what is stain solution for double heterostructure InP/InGaAsP?

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

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From: cbalane-at-wesleyan.edu
Date: Sun, 12 Feb 2006 01:38:59 -0600
Subject: [Microscopy] colloidal gold structure

Contents Retrieved from Microscopy Listserver Archives
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Hello listservers,
I realize that this might be a silly question but I was wondering if there
was anyone here who knows what the structure of colloidal gold is.

Thanks,
Carlo


--
Carlo Franco Bolivar Balane

Box 4058, 222 Church Street, or Wolfe Laboratory
Wesleyan University Station Rm. 157, HA Laboratories
Middletown, CT, 06459-4058 Wesleyan University



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7, 32 -- Subject: colloidal gold structure
7, 32 -- From: "Carlo Franco Balane-Bolivar" {cbalane-at-wesleyan.edu}
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From: neil-at-young8696.freeserve.co.uk
Date: Sun, 12 Feb 2006 06:20:41 -0600
Subject: [Microscopy] RE: colloidal gold structure

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

In general multiply-twinned fcc domains I would have thought. Although depending on particle size, very small colloids can have structure forbidden by conventional bulk crystallography such as a 5-fold icosahedral or decahedral arrangements, plenty of literature around, both experimental and simulations. I seem to remember passivation of the gold 'cluster' can force the resulting colloid into certain structures. Generally Au over 3-4nm in diameter is mt-fcc though.

Neil Young
Cluster Physics / Electron Microscopy
Nanoscale Physics Research Laboratory
School of Physics and Astronomy
University of Birmingham
UK



} Message Received: Feb 12 2006, 07:42 AM
} From: cbalane-at-wesleyan.edu
} To: neil-at-young8696.freeserve.co.uk
} Cc:
} Subject: [Microscopy] colloidal gold structure
}
}
}
}
} ----------------------------------------------------------------------------
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} Hello listservers,
} I realize that this might be a silly question but I was wondering if there
} was anyone here who knows what the structure of colloidal gold is.
}
} Thanks,
} Carlo
}
}
} --
} Carlo Franco Bolivar Balane
}
} Box 4058, 222 Church Street, or Wolfe Laboratory
} Wesleyan University Station Rm. 157, HA Laboratories
} Middletown, CT, 06459-4058 Wesleyan University
}
}
}
} ==============================Original Headers==============================
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From: kenconverse-at-qualityimages.biz
Date: Sun, 12 Feb 2006 14:39:26 -0600
Subject: [Microscopy] SEM x-ray peak ratios

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

Karl,
I hadn't seen any answer to your question so I'll give it a shot. For those
of us who grew up with Be windows, K bigger than L seems normal. However,
the norm for light element detectors is the opposite, at least if you
normally operate at 20kV or so. I'm wondering if you've had a long-standing
contamination problem with your light element window. Now that it's clean,
you may be seeing the correct ratio and may find that your light element
detection is also better.

The other thing to be certain of is that you are operating at the same kV as
you were earlier. Lower kV will favor lower energy peaks and higher kV will
favor higher energy peaks. It can be quite dramatic. Put a little graphite
on a piece of Cu tape and collect a spectrum from an area containing both.
Maintain a constant dead-time while collecting a spectrum at 30 kV and
another at about 2 kV. You'd never know it's the same sample.

Ken Converse
owner

QUALITY IMAGES
Servicing Scanning Electron Microscopes
Since 1981
474 So. Bridgton Rd.
Bridgton, ME 04009
207-647-4348
Fax 207-647-2688
kenconverse-at-qualityimages.biz
qualityimages.biz


-----Original Message-----
X-from: hagglundk1-at-nku.edu [mailto:hagglundk1-at-nku.edu]
Sent: Tuesday, January 24, 2006 10:33 AM
To: kenconverse-at-qualityimages.biz

I have a colleague who recently experienced some down time on his SEM.
The system went through repeated cycles of pump down, and venting and
was eventually left powered down while waiting on a computer
replacement.

After getting everything operating again, an oil film was found that had
built up on the x-ray detector thin window. This was initially causing
an unacceptable amount of background in the 0-3kV range of the x-ray
signal as well as an overall low count rate. The collimator was removed
and cleaned, which corrected the noise and count rate. They are in the
process of replacing the roughing lines and cleaning the chamber to
prevent further contamination. Now, when calibrating with a copper
standard, the ratio of the L line signal versus the K lines is
dramatically favoring the L. This was not the case before.

Any ideas why this might be happening and how to correct it?

Karl Hagglund
Biological Sciences, SC-102B
Northern Kentucky University, Nunn Drive
Highland Heights, KY 41099
859-572-5238
http://semlab.nku.edu


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From: George.Theodossiou-at-amcor.com.au
Date: Sun, 12 Feb 2006 18:58:09 -0600
Subject: [Microscopy] Nikon Microscope

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

Hi All,

A colleague has a Nikon SMZ-2T Microscope that he is trying to rescue. He
is after a copy of the manual. If anyone can help pls contact him at
hbeh-at-hotmail.com or via myself.

Thank you for your help

Regards
George



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From: gerd.leitinger-at-meduni-graz.at
Date: Mon, 13 Feb 2006 04:22:44 -0600
Subject: [Microscopy] EM: precautions when en bloc staining summary

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

Thank you everybody who replied to my question regarding safety
measures when working with uranyl acetate stained blocks.

To sum up the answers: The binding capacity of tissue to uranyl acetate is
apparently low and therefore very little (if any) radioactivity can escape
the blocks. However, when trimming I have been advised to wear gloves
and
a mask to prevent myself from inhaling chips from the specimen, and it
seems necessary that we collect the chips and dispose of them as
radioactive waste.

thank you

Gerd



Dr. Gerd Leitinger

Institut für Zellbiologie, Histologie und Embryologie
Medizinische Universität Graz
Harrachgasse 21
A-8010 Graz
Austria

Tel. ++43 316 380 4237
Fax. ++43 316 380 9625
Mailto: Gerd.Leitinger-at-meduni-graz.at



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From: aarti_harle-at-yahoo.co.in
Date: Mon, 13 Feb 2006 04:53:08 -0600
Subject: [Microscopy] confocal microscopy

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

Dear All

We are trying to viusalize infected erythrocytes
(malarial parasite) on confocal microscope by indirect
fluorescence without any fixation and washing is
carried out with PBS

The immages are not satisfactory because
fading/bleaching is fast though we are using % DABCO
in 50% glycerol and secondly lot of background noise.

Most of the literature shows the fixation with
ethanol/methenol but I would like to carry out the
fixation with paraformaldehyde for the obvious reason
and washing with MSM-PIPES with tris.

any suggestion????
or any body can describle the sample preparation for
infected erythrocytes using paraformaldehy as a
fixative and MSM PIPES as a washing buffer...

Does non fixation of specimen play any role in fast
bleaching???

Regards
Shrunali
Scientist
IMTECH, India



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From: phillipst-at-missouri.edu
Date: Mon, 13 Feb 2006 08:58:53 -0600
Subject: [Microscopy] Re: confocal microscopy

Contents Retrieved from Microscopy Listserver Archives
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You don't specify which fluorochrome you are using. If you are using FITC
or Rhodamine, that is one of the reasons you are getting rapid fading. The
Alexa488 and Alexa568 fluorochromes from Molecular Probes (Invitrogen) are
far superior in this regard - especially for confocal. Molecular Probes
also has a new anti-fade mounting medium called Prolong Gold but I don't
have enough experience at the moment to discuss its efficacy. Fixation
will not have any effect on bleaching but will contribute to background
noise (especially glutaraldehyde). Generally 2% PF is okay in regards to
background fluorescence and sometimes it is safe to add 0.1 - 02%
glutaraldehyde. Aldehyde fixatives may, however, interfere with your
antibody recognizing its epitope. good luck.

rote:



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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



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From: dljones-at-bestweb.net
Date: Mon, 13 Feb 2006 09:22:45 -0600
Subject: [Microscopy] Zeiss 940A

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I'm working on getting a Zeiss 940A up and running for my local high school. We
are working on a shoe-string budget as one may expect from a small public high
school.

I am looking for technical documentation for this instrument so that I can try
and get it functioning. If anyone has schematics or any other technical
documentation for instrument this I would greatly appreciate getting copies in
some way.

If anyone has a similar instrument that would be able to donate it to us for a
spare parts machine, we would come to pick it up. We are located in the Hudson
Valley.

I am willing to pay for copies if need be, as well as transportation costs.

Thank you in advance,

dj


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From: roncastellano-at-adelphia.net
Date: Mon, 13 Feb 2006 14:43:56 -0600
Subject: [Microscopy] AskAMicroscopist: Assistance to setup a JOEL JSM-35C SEM

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This Question was submitted to Ask-A-Microscopist by
(roncastellano-at-adelphia.net)
from
http://www.microscopy.org/Ask-A-Microscopist/Ask-A-Microscopist.html
on Monday, February 13, 2006 at 13:21:29
Remember to consider the Grade/Age of the student when considering the Question
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Email: roncastellano-at-adelphia.net
Name: Ron Castellano

Organization: Wolfram Analytical Labs

Education: Undergraduate College

Location: 924 Fords Corner Road, Nanty Glo, PA 15943

Title: Assistance to setup a JOEL JSM-35C SEM

Question: I'm a retired assay technician(precious metal analysis)
I have purchased an old SEM for private use in a small lab I am building here.
I need someone to set up and test unit at my site. The unit is said
to be operational when de-installed, it appears to be in good shape.
Are there any technicians, perhaps a graduate student, who can assist
me. Of course I am willing to pay any reasonable fee for this
service. Ron Castellano

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From: ken.blight-at-cancer.org.uk
Date: Wed, 15 Feb 2006 09:24:56 -0600
Subject: [Microscopy] SEM of Protozoa

Contents Retrieved from Microscopy Listserver Archives
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It is likely that I will need to view protozoa, in the SEM, in the near
future.Is there a special processing protocol or any problems to look out
for.


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From: eoptics-at-mcmaster.ca
Date: Wed, 15 Feb 2006 09:52:58 -0600
Subject: [Microscopy] TEM, Gatan 673 Wide Angle Camera, Looking for Spare Parts

Contents Retrieved from Microscopy Listserver Archives
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Good Day Readers:

I am in need of a working video card for a Gatan 673 Wide TV Camera,
vintage 1987.
This camera system is used on a Philips CM12 for teaching purposes.
The video card model is Schlumberger/Fairchild FAB-2-1-28 rev. F.

I will consider buying the controller if you do not wish to sell the card
separately.

If you wish you can contact me offline.

Thanks in advance,

Fred Pearson

*******************************************
Fred Pearson
McMaster University
Brockhouse Institute for Materials Research
ABB-B145
1280 Main Street West
Hamilton ON. Canada
L8S 4M1

email:eoptics-at-mcmaster.ca
phone: (905) 525-9140 ext.24609
fax: (905) 521-2773
*******************************************

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From: wim5-at-lehigh.edu
Date: Wed, 15 Feb 2006 12:43:10 -0600
Subject: [Microscopy] JEOL JSM-6300f FEGSEM for sale

Contents Retrieved from Microscopy Listserver Archives
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Lehigh University, Center for Advanced Materials and Nanotechnology,
Bethlehem, PA is offering for sale a JEOL-6300F FEGSEM which will be
decommissioned in mid-March 2006. The Microscope includes an Oxford/ISIS
thin window EDS system, JEOL dry airlock and ARC64 digital imaging
system, solid state backscatter detector, IR chamberview camera. Asking
price: $60K Interested parties can contact Dr. Chris Kiely at
chk5-at-lehigh.edu.

For technical questions regarding the microscope please contact Bill
Mushock wim5-at-lehigh.edu

--




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From: tonygr-at-MIT.EDU
Date: Wed, 15 Feb 2006 13:26:25 -0600
Subject: [Microscopy] Transfer of used equipment originally bought with US funds

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

I'm writing this as a response to Chris Kiely's post, but it is a
very general point that probably bears repeating from time to time.

If a piece of equipment has been purchased by US Government funds
(regardless of agency), then it is not permissible to use other US
Government funds to re-purchase the same piece of equipment
later. This is true even if title to the equipment has been passed
to the holding instrumentation.

To use the specific example of the Lehigh SEM, if it was purchased
through a government grant, I would not be able to buy it from them,
regardless of how much use it would be to me, because the only funds
I have available (at least for the purposes of my illustration!) are
NSF funds. This is a limitation enforced on me, as the spender of
Government funds, rather than on the seller (for they are -
presumably - selling their own property), but it does mean that I
need to know up front whether the instrument was purchased with any
US Government funds (even if other funds were used as well) before I
can decide whether I might be interested in it. I, and my
institution, would be in major hot water should an auditor discover I
had used government funds in this way (I suspect that here at MIT our
internal systems would catch this before the sale went through, but
that may not be true everywhere).

Tony Garratt-Reed.



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Anthony J. Garratt-Reed, M.A., D.Phil.
MIT Room 13-1027
77 Massachusetts Avenue
Cambridge, MA 02139-4307
USA

Tel: 617-253-4622
Fax: 617-258-6478
************************************************


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From: zaluzec-at-microscopy.com
Date: Wed, 15 Feb 2006 14:59:46 -0600
Subject: [Microscopy] Administrivia: please read the FAQ

Contents Retrieved from Microscopy Listserver Archives
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Colleagues

Just a reminder to please read the FAQ if you not sure about a posting.

Posting of equipment/items for sale on the Listserver is against our rules.
You should use the MSA Surplus Equipment site for that function.

Nestor
Your Friendly Neighborhood SysOp

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From: scanning-at-fams.org
Date: Thu, 16 Feb 2006 10:16:33 -0600
Subject: [Microscopy] SCANNING 2006 Abstract Deadline Extended to March 1

Contents Retrieved from Microscopy Listserver Archives
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Dear Listers,

Please note the following:

ABSTRACT DEADLINE EXTENDED:

Because it's abstract time for both SCANNING 2006 and M&M, we know
there is time pressure on all contributors. We are therefore extending
the abstract deadline for SCANNING 2006 to March 1. Abstracts will
appear in the Program Issue of SCANNING (March-April Issue) if received
by March 1. Thank you.

For current program information and to download the meeting and hotel
registration forms, please visit www.scanning.org.

Best regards,


Phaedra McGuinness
Managing Editor
SCANNING, The Journal of Scanning Microscopies
P.O. Box 485
Mahwah, NJ 07430
Tel: (201) 818-1010 * Fax: (201) 818-0086 *email: scanning-at-fams.org *
Web: www.scanning.org


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From: grmitch-at-netzero.com
Date: Thu, 16 Feb 2006 14:48:15 -0600
Subject: [Microscopy] AskAMicroscopist: teaching a unit on microscopic life to 5th

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This Question was submitted to Ask-A-Microscopist by (grmitch-at-netzero.com)
from
http://www.microscopy.org/Ask-A-Microscopist/Ask-A-Microscopist.html
on Wednesday, February 15, 2006 at 20:08:01
Remember to consider the Grade/Age of the student when considering the Question
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Email: grmitch-at-netzero.com
Name: Linda Mitchell

Organization: BPS Environmental Center

Education: K-8 Grade Grammar School

Location: Birmingham, Michigan USA

Title: Microorganisms!

Question: I will be teaching a unit on microscopic life to 5th
graders next month, March (quite a cold month here in Michigan). My
question for you:
What is the best way to keep my microorganisms alive throughout the
program? We obtain our samples directly from the 10 acres of the
property here at the nature center. One of the samples is from our
pond area and the second from the swamp woods area. We have no
difficulty in finding a healthy sample, yet the problem that we often
encounter is keeping them alive. We have supplies - lab pans,
microscopes, even an aquarium that is our "indoor pond" when the
water ices over. Do you have any feeding, climate tips that would
help in these areas? This is a program that is enjoyed by both the
staff and students - they are so amazed by what they find. Thanks for
your input.

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

==============================Original Headers==============================
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8, 13 -- From: grmitch-at-netzero.com (by way of Ask-A-Microscopist)
8, 13 -- Subject: AskAMicroscopist: teaching a unit on microscopic life to 5th
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From: zaluzec-at-microscopy.com
Date: Thu, 16 Feb 2006 14:52:48 -0600
Subject: [Microscopy] MM2006 Server OverLoad - Deadline extended

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

Colleagues

I'm out of the country right now, but I have been getting multiple
questions about problems with the MM2006 server for submitting
papers.

Please note that there was a HUGE spike in the paper submissions
and the server is overloaded. As a consequence the MM2006
Executive committee has extended the deadline for paper submissions.

A note to this effect is appended below.

Nestor
Your Friendly Neighborhood SysOp (in Australia for the moment).


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


The paper submission deadline for M&M 2006 in Chicago has been
extended two days until 5:00pm Pacific Standard Time on Friday
February 17, 2006. Due to an ongoing problem with the server today we
have decided to extend the deadline to allow everyone the opportunity
to submit their papers. Please try to access the paper submission
site {http://bono.cup.org/} http://bono.cup.org/ later to register and
submit your paper. Our sincerest apologies for any problems this may
have caused.

Thanks,
Paul Kotula
Microscopy &Microanalysis 2006 Program Chair

Paul G. Kotula, Ph.D.
Principal Member of Technical Staff
Materials Characterization Department
Sandia National Laboratories
PO Box 5800, MS 0886
Albuquerque, NM 87185-0886

ph:(505) 844-8947

==============================Original Headers==============================
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From: sue.tyler-at-noaa.gov
Date: Thu, 16 Feb 2006 14:56:50 -0600
Subject: [Microscopy] viaWWW: Glass Knife Breaker

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Email: sue.tyler-at-noaa.gov
Name: Sue Tyler

Organization: NOAA

Title-Subject: [Filtered] Glass knife breaker

Question: I read the recent string on the glass knife makers and I
was interested in the conclusion. We are interested in purchasing
either the GKM or the Leica KMR. Any pros or cons would be
appreciated.

Regards-
Sue

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

==============================Original Headers==============================
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7, 12 -- From: sue.tyler-at-noaa.gov (by way of MicroscopyListserver)
7, 12 -- Subject: viaWWW: Glass Knife Breaker
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From: diller-at-stefan-diller.com
Date: Thu, 16 Feb 2006 14:57:31 -0600
Subject: [Microscopy] viaWWW: TN2000 EDS System

Contents Retrieved from Microscopy Listserver Archives
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Email: diller-at-stefan-diller.com
Name: Stefan Diller

Organization: Scientific Photography

Title-Subject: [Filtered] Tracor Northern TN2000 EDS system

Question: Dear All,
is anybody out there knowing some details or having some images
available of an 1991 Tracor Northern TN2000 EDS system?

Is there still a possiblity to get service in Germany?
Is there any possibility to get the EDS data out of the analyzer into
a PC for saving, printing, emailing?
What is the latest version of software for the TN2000?

Please reply offline, if convenient...
I will do a summarizing for the list...

Best regards,
Stefan

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

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9, 12 -- From: diller-at-stefan-diller.com (by way of MicroscopyListserver)
9, 12 -- Subject: viaWWW: TN2000 EDS System
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From: soleimanij-at-tbzmed.ac.ir
Date: Thu, 16 Feb 2006 14:57:52 -0600
Subject: [Microscopy] viaWWW: cutting polymer

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Email: soleimanij-at-tbzmed.ac.ir
Name: Jafar Soleimani Rad

Organization: University

Title-Subject: [Filtered] cutting a polymer

Question: We are having problem with cutting a polymer, Acrylonitril
Butadien Styrene (ABS.
After embedding in resin it dose not stick to it and becomes
separated when trying to cut with ultramicrotom. It is also
impossible to cut paraffin blocks. I would appreciate if you guide me
with your experiences in this matter.

Sincerely

JSRad


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

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9, 12 -- Subject: viaWWW: cutting polymer
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From: b.farwell-at-unf.edu
Date: Thu, 16 Feb 2006 14:59:17 -0600
Subject: [Microscopy] viaWWW: AFM controller connection problems

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Email: b.farwell-at-unf.edu
Name: Brenton

Organization: University of N. Florida (student)

Title-Subject: [Filtered] AFM controller connection problems

Question: We are using a Molecular Imaging AFM and controller and are
using "MI Metrology Series 2000 AFM System" program to interface to
the AFM controller. The computer is having connection issues with
the controller. Cables have been checked, and the setup disk has run
on the controller, it gives 4 beeps, however the computer still can't
connect. Ping gives no response. Anybody have any ideas?
Thanks very much
Brenton

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

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From: jcrum-at-ncmir.ucsd.edu
Date: Thu, 16 Feb 2006 15:52:20 -0600
Subject: [Microscopy] Wanted: Balzers MED 010 Manual

Contents Retrieved from Microscopy Listserver Archives
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Hello,

I recently acquired a used Balzers MED 010 of unknown vintage. I am in
need of the User Manual, or any such literature. I would like to get
this unit back to operating condition.

Thanks,

John Crum
NCMIR
UCSD

==============================Original Headers==============================
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From: andy-at-psemdoctor.com
Date: Thu, 16 Feb 2006 17:20:44 -0600
Subject: [Microscopy] Looking for used PSEM

Contents Retrieved from Microscopy Listserver Archives
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I am looking for a used PSEM (or parts) to purchase. Preferably this system
would be the original model manufactured by RJ Lee Instruments in the mid to
late 90’s. Anyone that can help me out, please email directly to
andy-at-psemdoctor.com.
Thank you,

Andrew L. Zobel
 
PSEMDOCTOR, LLC
117 Bryant Dr.
Pittsburgh, PA 15235
Tel. 412-215-8906
Fax 412-241-3598
WWW.PSEMDOCTOR.COM




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From: ech-at-interchange.ubc.ca
Date: Thu, 16 Feb 2006 19:56:45 -0600
Subject: [Microscopy] Re: viaWWW: Glass Knife Breaker

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

Hi Sue
We have the Leica glass knife breaker since 2001. We are very happy
with it. Reliable, good knives! Our users like it in preference to
our old workhorse LKB knife breakers.
Elaine

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


--
Dr. Elaine Humphrey
Director, BioImaging Facility
President, Microscopy Society of Canada (2003-2005)
University of British Columbia
6270 University Blvd, mail-stop Botany
Vancouver, BC
CANADA, V6T 1Z4
Phone: 604-822-3354
FAX: 604-822-6089
e-mail: ech-at-interchange.ubc.ca
website: www.emlab.ubc.ca

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From: jeff-at-metallography.com
Date: Fri, 17 Feb 2006 08:43:23 -0600
Subject: [Microscopy] Seeking manual for B&L Research I metallograph

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

Listers,

A colleague is looking for the instruction manual for a Bausch & Lomb
Research I metallograph. If anybody can help him in this quest please
respond directly to him, Gregory Dexter at greg-at-met-sol.com .

Thanks,

Jeff Stewart
Metallographic Laboratory Manager
Stern-Leach Company
49 Pearl Street
Attleboro, MA 02703
508-222-7400 x1329


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From: dsherman-at-purdue.edu
Date: Fri, 17 Feb 2006 11:51:22 -0600
Subject: [Microscopy] Microscope cooling lines

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Listers,

For years we have been wondering where the dark material was coming from
that accumulated in our water filters. These are filters in the closed
circuit lines between our microscopes and water recirculating units. The
lines are mainly in the ceiling or totally insulated as they run down the
walls in the scope rooms. Imaging our surprise when we went to do a minor
repair on one and watched as the plumber removed a regulator and inserted a
piece of galvanized pipe.

Apparently when the building was built (20 years ago), the contractor
used galvanized pipe when copper had been specified. As it was hidden, we
did not know about the switch. All visible lines hooking up the chiller
compressor cooling with building water were copper.

Well now these galvanized lines are really breaking down and clogging the
water pumps. We intend to replace all but were questioning whether it would
be best to replace with copper or PVC piping. Any suggestions?

One concern was whether there would be the need to acid clean these lines in
the future (this is done routinely to the compressor lines to remove mineral
build-up). Since it is closed circuit, we should not accumulate large
amounts of minerals even though tap or deionized water will be used for the
system. We also can control algae growth with chemicals. Any suggestions on
this and should this dictate which material is used for the pipes?

Debby

Debby Sherman, Manager Phone: 765-494-6666
Life Science Microscopy Facility FAX: 765-494-5896
Purdue University E-mail: dsherman-at-purdue.edu
S-052 Whistler Building
170 S. University Street
West Lafayette, IN 47907
http://www3.agriculture.purdue.edu/microscopy


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8, 21 -- Subject: Microscope cooling lines
8, 21 -- From: Debby Sherman {dsherman-at-purdue.edu}
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From: wpchan-at-u.washington.edu
Date: Fri, 17 Feb 2006 12:15:44 -0600
Subject: [Microscopy] manual for Denton DV-502A

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

Hi

I wonder if anyone has a manual for the Denton DV-502A vacuum evaporator
that I can borrow. Ours was lost during moving to storage and now I have
to set it up again. It would be nice to make sure I set it up correctly.
Denton can supply the manual for $250 so I am looking at other less
expensive ways first. Thanks a lot!

--
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: bozzola-at-siu.edu
Date: Fri, 17 Feb 2006 12:57:04 -0600
Subject: [Microscopy] Re: Microscope cooling lines

Contents Retrieved from Microscopy Listserver Archives
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Debby,

For our new IMAGE building, I specified PVC piping for the closed
circ loop between the water chillers and scopes. We still notice a
greenish sludge building up in the water filters (takes about 6-8
months to become significant) but I am certain that this is coming
from the EM (copper cooling coils and iron
connections---} electrolytic reaction). The EM service people told us
that if we ever used acid to clean the lines that they would no
longer warranty the microscope. The PVC lines are perfectly clean.

JB




} For years we have been wondering where the dark material was coming from
} that accumulated in our water filters. These are filters in the closed
} circuit lines between our microscopes and water recirculating units. The
} lines are mainly in the ceiling or totally insulated as they run down the
} walls in the scope rooms. Imaging our surprise when we went to do a minor
} repair on one and watched as the plumber removed a regulator and inserted a
} piece of galvanized pipe.
}
} Apparently when the building was built (20 years ago), the contractor
} used galvanized pipe when copper had been specified. As it was hidden, we
} did not know about the switch. All visible lines hooking up the chiller
} compressor cooling with building water were copper.
}
} Well now these galvanized lines are really breaking down and clogging the
} water pumps. We intend to replace all but were questioning whether it would
} be best to replace with copper or PVC piping. Any suggestions?
}
} One concern was whether there would be the need to acid clean these lines in
} the future (this is done routinely to the compressor lines to remove mineral
} build-up). Since it is closed circuit, we should not accumulate large
} amounts of minerals even though tap or deionized water will be used for the
} system. We also can control algae growth with chemicals. Any suggestions on
} this and should this dictate which material is used for the pipes?
}
} Debby
}
} Debby Sherman, Manager Phone: 765-494-6666
} Life Science Microscopy Facility FAX: 765-494-5896
} Purdue University E-mail: dsherman-at-purdue.edu
} S-052 Whistler Building
} 170 S. University Street
} West Lafayette, IN 47907
} http://www3.agriculture.purdue.edu/microscopy
}
}
} ==============================Original Headers==============================
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} 8, 21 -- Subject: Microscope cooling lines
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} 8, 21 -- To: "message to: MSA list" {microscopy-at-microscopy.com}
} 8, 21 -- Message-ID: {C01B7748.D5AB%dsherman-at-purdue.edu}
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--
##############################################################
John J. Bozzola, Ph.D., Director
I.M.A.G.E. (Integrated Microscopy & Graphics Expertise)
750 Communications Drive - MC 4402
Southern Illinois University
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Phone: 618-453-3730
Email: bozzola-at-siu.edu
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From: pgrover-at-bilbo.bio.purdue.edu
Date: Fri, 17 Feb 2006 13:19:13 -0600
Subject: [Microscopy] re: microscope cooling lines

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

Debby,

Another material you may wish to consider is called PEX, which is a
cross-linked polyethylene. I don't know too much about its characteristics,
except that it's very smooth inside, which should retard crud accumulation
and it's more opaque than white pvc. It may be worth checking out.

Paul


--------------------------------------------------------------------------
Famous Last Words Department: "I did not get my Spaghetti-O's, I got
spaghetti. I want the press to know this."
~~ Thomas J. Grasso, d. March 20, 1995 Executed by injection, Oklahoma.



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7, 21 -- Subject: re: microscope cooling lines
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From: dljones-at-bestweb.net
Date: Fri, 17 Feb 2006 13:24:27 -0600
Subject: [Microscopy] Re: Microscope cooling lines

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

Debbie,

It would be good to know what is the operating temperature, pressure, flow rate
and the characteristics of the water feed for make-up.

There is no material in-compatibility between copper piping and PVC piping. They
can be used in the same water curcuit.

If you want to know if there is a problem using copper, you can take some of the
water out of the system and see how much copper is present. You need to have a
base line of copper from the water source, so you should take a water sample
from the source also. There is likely not a problem, it depends upon your water
chemistry. Knowing your water chemistry is fundamental to knowing what piping is
preferred.

You may have building code restrictions regarding PVC piping that is hidden,
which may be the reason the orginal contractor put in galvanized piping. You
will have to look at what codes apply to where you are.

Regarding acid cleaning, you should know what your deposits are in your piping
before deciding how to clean them. As an FYI, copper will generally corrode at
pH's below about 6.3 or so. There are low pH cleaners that can be used with
copper, but they contain corrosion inhibitors.

dj

On Fri, 17 Feb 2006 dsherman-at-purdue.edu wrote:

}
}
}
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
} To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
} On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} ----------------------------------------------------------------------------
}
} Listers,
}
} For years we have been wondering where the dark material was coming from
} that accumulated in our water filters. These are filters in the closed
} circuit lines between our microscopes and water recirculating units. The
} lines are mainly in the ceiling or totally insulated as they run down the
} walls in the scope rooms. Imaging our surprise when we went to do a minor
} repair on one and watched as the plumber removed a regulator and inserted a
} piece of galvanized pipe.
}
} Apparently when the building was built (20 years ago), the contractor
} used galvanized pipe when copper had been specified. As it was hidden, we
} did not know about the switch. All visible lines hooking up the chiller
} compressor cooling with building water were copper.
}
} Well now these galvanized lines are really breaking down and clogging the
} water pumps. We intend to replace all but were questioning whether it would
} be best to replace with copper or PVC piping. Any suggestions?
}
} One concern was whether there would be the need to acid clean these lines in
} the future (this is done routinely to the compressor lines to remove mineral
} build-up). Since it is closed circuit, we should not accumulate large
} amounts of minerals even though tap or deionized water will be used for the
} system. We also can control algae growth with chemicals. Any suggestions on
} this and should this dictate which material is used for the pipes?
}
} Debby
}
} Debby Sherman, Manager Phone: 765-494-6666
} Life Science Microscopy Facility FAX: 765-494-5896
} Purdue University E-mail: dsherman-at-purdue.edu
} S-052 Whistler Building
} 170 S. University Street
} West Lafayette, IN 47907
} http://www3.agriculture.purdue.edu/microscopy
}
}
} ==============================Original Headers==============================
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} 8, 21 -- Received: from exchange.purdue.edu (1061exfe02.adpc.purdue.edu [128.210.63.223])
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} 8, 21 -- Subject: Microscope cooling lines
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From: tivol-at-caltech.edu
Date: Fri, 17 Feb 2006 13:36:47 -0600
Subject: [Microscopy] Re: Microscope cooling lines

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


On Feb 17, 2006, at 9:51 AM, dsherman-at-purdue.edu wrote:

} For years we have been wondering where the dark material was coming
} from
} that accumulated in our water filters. These are filters in the closed
} circuit lines between our microscopes and water recirculating units.
} The
} lines are mainly in the ceiling or totally insulated as they run down
} the
} walls in the scope rooms. Imaging our surprise when we went to do a
} minor
} repair on one and watched as the plumber removed a regulator and
} inserted a
} piece of galvanized pipe.
}
} Apparently when the building was built (20 years ago), the
} contractor
} used galvanized pipe when copper had been specified. As it was
} hidden, we
} did not know about the switch. All visible lines hooking up the
} chiller
} compressor cooling with building water were copper.
}
} Well now these galvanized lines are really breaking down and clogging
} the
} water pumps. We intend to replace all but were questioning whether it
} would
} be best to replace with copper or PVC piping. Any suggestions?
}
} One concern was whether there would be the need to acid clean these
} lines in
} the future (this is done routinely to the compressor lines to remove
} mineral
} build-up). Since it is closed circuit, we should not accumulate large
} amounts of minerals even though tap or deionized water will be used
} for the
} system. We also can control algae growth with chemicals. Any
} suggestions on
} this and should this dictate which material is used for the pipes?
}
Dear Debby,
If you intend to clean the lines with acid, I suggest PVC, since Cu
can be etched at low pH. In addition to floating some dichlorophene
for algal control, we add a corrosion inhibitor. We have been using a
Mo-based formula, which was available from Aqua Labs on the East coast
and from Skasol on the West coast, so find a distributor in your area.
I think that either Aqua or Skasol would be able to give you that info.
Yours,
Bill Tivol, PhD
EM Scientist and Manager
Cryo-Electron 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: tivol-at-caltech.edu
Date: Fri, 17 Feb 2006 13:42:58 -0600
Subject: [Microscopy] microscope cooling lines

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


On Feb 17, 2006, at 11:19 AM, pgrover-at-bilbo.bio.purdue.edu wrote:

} Another material you may wish to consider is called PEX, which is a
} cross-linked polyethylene. I don't know too much about its
} characteristics,
} except that it's very smooth inside, which should retard crud
} accumulation
} and it's more opaque than white pvc. It may be worth checking out.
}
Dear Paul,
PEX sounds like a better material than PVC. It should be essentially
inert--I expect that it will withstand concentrated bases and acids and
would be unaffected by organic solvents (not that one would want to run
those through the scope lines).
Yours,
Bill Tivol, PhD
EM Scientist and Manager
Cryo-Electron Microscopy Facility
Broad Center, Mail Code 114-96
California Institute of Technology
Pasadena CA 91125
(626) 395-8833
tivol-at-caltech.edu


==============================Original Headers==============================
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From: dljones-at-bestweb.net
Date: Fri, 17 Feb 2006 13:43:51 -0600
Subject: [Microscopy] Microscope cooling lines

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

JB,

As an FYI, if you have copper based and iron based materials mixed in the same
system, the iron will corrode preferentially through galvanic coupling, not the
copper. In fact, the iron becomes a sacrifical anode protecting the copper from
corrosion. Any time you have those two materials in the same system, you should
have dielectric couplings between the two or you will actively corrode the
ferrous based material.

If you are having a copper corrosion problem, you should be looking elsewhere
for the cause...

dj

On Fri, 17 Feb 2006 bozzola-at-siu.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
} ----------------------------------------------------------------------------
}
} Debby,
}
} For our new IMAGE building, I specified PVC piping for the closed
} circ loop between the water chillers and scopes. We still notice a
} greenish sludge building up in the water filters (takes about 6-8
} months to become significant) but I am certain that this is coming
} from the EM (copper cooling coils and iron
} connections---} electrolytic reaction). The EM service people told us
} that if we ever used acid to clean the lines that they would no
} longer warranty the microscope. The PVC lines are perfectly clean.
}
} JB
}


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From: jecalvin-at-vassar.edu
Date: Fri, 17 Feb 2006 14:40:00 -0600
Subject: [Microscopy] Re: Microscope cooling lines

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

Along these lines, I noticed a lot more copper leaching into the
lines when I used deionized water than regular or filtered tap water.

Secondly, Why hasn't anyone suggested using antifreeze? Antifreeze
intended for cars should have corrosion inhibitors and should be
suitable for this purpose.

I have still gotten green particles regardless of whether antifreeze
was used or not and have regularly made it a practice to clean the
filters in the lines once a semester. Jerry Calvin



At 1:46 PM -0600 2/17/06, dljones-at-bestweb.net wrote:
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America


--
****************************
Jerry G. Calvin
Science Support Technician
Box 0731 Biology Department
Vassar College
124 Raymond Avenue
Poughkeepsie, NY 12604-0731

(845) 437-7423 - Office
(845) 437-7424 - Confocal Room
FAX: (845) 437-7315
E-Mail: jecalvin-at-vassar.edu

==============================Original Headers==============================
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From: davilla-at-4pi.com
Date: Fri, 17 Feb 2006 15:17:27 -0600
Subject: [Microscopy] Re: microscope cooling lines

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

} PEX sounds like a better material than PVC. It should be essentially

Beware that PEX will degrade over time in sunlight (ultraviolet).
That said, I have PEX in my house instead of Cu, works well and is
easy to run, however all transitions through the walls are Cu
fittings so the PEX stays in the dark.

Scott
--
-----------------------------------------------------------------------
Scott D. Davilla Phone: 919 489-1757 ext 13 (tel)
4pi Analysis, Inc. Fax: 919 489-1487 (fax)
3500 Westgate Drive, Suite 403 email: davilla-at-4pi.com
Durham, North Carolina 27707-2534 web: http://www.4pi.com


==============================Original Headers==============================
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From: dljones-at-bestweb.net
Date: Fri, 17 Feb 2006 15:21:32 -0600
Subject: [Microscopy] Re: Microscope cooling lines

Contents Retrieved from Microscopy Listserver Archives
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Jerry,

Yes, it is not unusual to find more copper leaching into the lines with
deionized water than tap water. It does depend upon the tap water chemistry.

I would not recommend antifreeze for cars, however, I would recommend to use
HVAC grade propylene glycol. That would include inhibitors to protect the metals
in the system and it includes various stablizers for the glycol. I guess if
there are glycols for cars that are propylene glycol based with inhibitors,
those may be OK. I most certainly would not use ethylene glycol based
antifreeze.

I would ask you what kind of antifreeze you are using and have you done an EDS
spectrum of your green deposit from you filters? I would think further
discussion of your specifics may best be addressed off the list...

dj

On Fri, 17 Feb 2006, Jerry Calvin wrote:

} Along these lines, I noticed a lot more copper leaching into the lines when
} I used deionized water than regular or filtered tap water.
}
} Secondly, Why hasn't anyone suggested using antifreeze? Antifreeze intended
} for cars should have corrosion inhibitors and should be suitable for this
} purpose.
}
} I have still gotten green particles regardless of whether antifreeze was used
} or not and have regularly made it a practice to clean the filters in the
} lines once a semester. Jerry Calvin
}
}
}
} At 1:46 PM -0600 2/17/06, dljones-at-bestweb.net wrote:
} } ----------------------------------------------------------------------------
} } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
} } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
} } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} } ----------------------------------------------------------------------------
} }
} } JB,
} }
} } As an FYI, if you have copper based and iron based materials mixed in the
} } same
} } system, the iron will corrode preferentially through galvanic coupling, not
} } the
} } copper. In fact, the iron becomes a sacrifical anode protecting the copper
} } from
} } corrosion. Any time you have those two materials in the same system, you
} } should
} } have dielectric couplings between the two or you will actively corrode the
} } ferrous based material.
} }
} } If you are having a copper corrosion problem, you should be looking
} } elsewhere
} } for the cause...
} }
} } dj
} }
} } On Fri, 17 Feb 2006 bozzola-at-siu.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
} } } ----------------------------------------------------------------------------
} } }
} } } Debby,
} } }
} } } For our new IMAGE building, I specified PVC piping for the closed
} } } circ loop between the water chillers and scopes. We still notice a
} } } greenish sludge building up in the water filters (takes about 6-8
} } } months to become significant) but I am certain that this is coming
} } } from the EM (copper cooling coils and iron
} } } connections---} electrolytic reaction). The EM service people told us
} } } that if we ever used acid to clean the lines that they would no
} } } longer warranty the microscope. The PVC lines are perfectly clean.
} } }
} } } JB
} } }
} }
} }
} } ==============================Original
} } Headers==============================
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} } 7, 25 -- To: bozzola-at-siu.edu
} } 7, 25 -- cc: Microscopy-at-microscopy.com
} } 7, 25 -- Subject: Re: [Microscopy] Re: Microscope cooling lines
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} } Headers==============================
}
}
} --
} ****************************
} Jerry G. Calvin
} Science Support Technician
} Box 0731 Biology Department
} Vassar College
} 124 Raymond Avenue
} Poughkeepsie, NY 12604-0731
}
} (845) 437-7423 - Office
} (845) 437-7424 - Confocal Room
} FAX: (845) 437-7315
} E-Mail: jecalvin-at-vassar.edu
}


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From: bagnell-at-med.unc.edu
Date: Fri, 17 Feb 2006 16:01:57 -0600
Subject: [Microscopy] Floaters

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

A colleague asked me why she is so much more aware of the
floaters in her eyes when she is using the microscope (or the
telescope) than at other times. I assured her that it wasn't just
her, it happens to a lot of us. But why floaters are so much more
noticeable when looking in the microscope I do not know? Any
suggestions?

Bob
--
C. Robert Bagnell, Jr., Ph.D.
Professor and
Director, Microscopy Services Laboratory
Department of Pathology and Laboratory Medicine
University of North Carolina at Chapel Hill
Chapel Hill, NC 27599
phone 919-966-2413
fax 919-966-6718
e-mail bagnell-at-med.unc.edu
web http://www.med.unc.edu/microscopy

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From: jfactor-at-ns.purchase.edu
Date: Fri, 17 Feb 2006 16:02:30 -0600
Subject: [Microscopy] Re: Microscope cooling lines

Contents Retrieved from Microscopy Listserver Archives
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My service engineer recommends neither tap nor distilled water, but
rather bottled spring water. Has anyone yet mentioned the possibility
that green sludge in the filter might be algae growing in the cooling
water or the cooling unit?
--Jan Factor

---------------------------------------
Jan Robert Factor, Ph.D.
Professor of Biology
---------------------------------------
Natural Sciences
Purchase College, State University of New York
735 Anderson Hill Rd.
Purchase, NY 10577
USA
---------------------------------------
Office Tel: 914-251-6659
Office Fax: 914-251-6635
E-mail: jfactor-at-ns.purchase.edu
or- jan.factor-at-purchase.edu
---------------------------------------



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From: leis-at-biochem.mpg.de
Date: Fri, 17 Feb 2006 16:07:44 -0600
Subject: [Microscopy] viaWWW: microtome section thickness

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Email: leis-at-biochem.mpg.de
Name: Andrew Leis

Organization: MPI for Biochemistry

Title-Subject: [Filtered] microtome section thickness

Question: Can anybody provide an explanation concerning the
refractive index -dependency of the interference colours seen in
charts for estimating the thickness of ultrathin sections? The small
print in such charts usually reads "valid for refractive index of ca.
1.5", which is fine for methacrylates and similar embedding media,
but what type of shift (if significant) would be observed for a lower
refractive index, say 1.3? I am not sure whether the usual charts are
valid for the interference colours given by cryosections.

Any assistance would be appreciated.

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

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From: milton.charlton-at-utoronto.ca
Date: Fri, 17 Feb 2006 16:08:13 -0600
Subject: [Microscopy] viaWWW: NTA-nanogold for His tagged protein in EM?

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Email: milton.charlton-at-utoronto.ca
Name: Milton Charlton

Organization: University of Toronto

Title-Subject: [Filtered] NTA-nanogold for His tagged protein in EM?

Question: Has anynone had success using NTA-nanogold to disclose the
location of His-tagged proteins by electron microscopy? I would like
to attach the nanogold this way before injecting the protein into a
cell. An alternative would be to apply gold labelled primary
anti-His after fixation and permeabilization.

Thanks

Milton Charlton
University of Toronto

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From: johnf-at-geology.wisc.edu
Date: Fri, 17 Feb 2006 16:21:54 -0600
Subject: [Microscopy] student scholarships: NIST/MAS Particle Workshop April 2006

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Please announce and distribute to students:

4 Student Scholarships ($500) Available for support for attending and
presenting a paper/poster

PARTICLE WORKSHOP 2006

A Joint NIST / Microbeam Analysis Society
Special Topics Workshop

Dates: April 24th - 26th 2006

Location: NIST, Gaithersburg, MD

The analysis of microscopic particles represents a measurement
challenge in a diverse range of scientific, technological and
environmental disciplines. Examples of areas of interest include
nanoscale particles as building blocks for nanotechnology,
contamination in high technology manufacturing, trace forensic
detection for homeland security, and environmental sampling and
monitoring. Characterization of particle specimens is complicated by
mass, volume and morphological effects that cause well established
bulk techniques to break down. In addition, while the measurement
types under consideration at this workshop are performed on a
particle-by-particle basis, the properties of interest are often
statistical measures of populations of similar particles.

This workshop will focus on the challenges that face industrial and
governmental application of microscopic particle measurement
techniques. The practical tone of the meeting will set by speakers
representing various industries and governmental organizations who
will provide insight into their particle measurement requirements.
These speakers will be followed by sessions that focus on sample
preparation and statistical experimental design and established &
emerging measurement techniques. Instrumentation to be discussed
include SEM/EDS, AEM, TOF/SIMS, optical, FIB and scanned probe.
Each of these sessions will be concluded by a round table discussion
in which attendees are encouraged to contribute.

Registration Deadline: March 31, 2006

Students interested in applying for one of the 4 student scholarships
should immediately contact

Nicholas W. M. Ritchie
NIST Microanalysis Research Group
100 Bureau Drive STOP 8371
Gaithersburg, MD 20899-8371
301-975-3929
nicholas.ritchie-at-nist.gov


There is no registration fee: the workshop is free to all attendees.
However, space is limited and pre-registration is required by NIST
Security for entry onto the NIST campus. This rule is strictly
enforced.

Full information at
www.cstl.nist.gov/div837/Division/meetings/particleworkshop/particle.htm

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From: bfoster-at-mme1.com
Date: Fri, 17 Feb 2006 16:31:50 -0600
Subject: [Microscopy] Re: Floaters

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

Hi, Bob

It has to do with the physics of simple pinhole optics. Essentially,
when you are just in the right focal plane, you are doing an "entopic
exam" of your eye. You can also reproduce the experiment by putting
a small hole (about 1/8") into a piece of cardboard (1/2 of a file
folder) and staring through it at a neutral surface (blank wall; sky,
if not too bright). Adjust the distance between the card and your
eye and, at the right distance, you will see the internal structure.

I've had an interesting array of tiny cataracts for over 25 years and
keep track of their position and size using this method.

Hope this is helpful.

Best regards,
Barbara Foster

Microscopy/Microscopy Education
313 S Jupiter Rd, Suite 100
Allen, TX 75002
P: 972-954-8011
W: www.MicroscopyEducation.com

P. S.
Need a good general reference or light microscopy text for the Spring
semester? Call us today to learn more about "Optimizing LIght
Microscopy". Copies still available through MME... even for
class-room lots ... and we give quantity discounts. Call Ken Piel at
(972)954-8011.



At 04:05 PM 2/17/2006, bagnell-at-med.unc.edu wrote:



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From: gary-at-gaugler.com
Date: Fri, 17 Feb 2006 16:58:51 -0600
Subject: [Microscopy] Microscope cooling lines

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

I had the same sludge problem about 1.5 years ago in
a new Haskris chiller. Haskris recommends using only
distilled water. That lasted about three months and
the slime appeared. It was a combination of algae and
small particles. Dumped the water and replaced with
new distilled water and Skasol to flush. Then, new
distilled water and one half liter of Hexid A4 from
Applied Thermal Control Ltd. UK as supplied by SEM
service tech. Chiller seized after about three months.
Post mortem indicated that the impeller blades failed.
Most likely due to misalignment of motor and pump.

Haskris replaced motor and pump assembly. Fluid was
drained and replaced with distilled water and ethylene
glycol (.5G to 4.5G distilled water). Filters were
changed and no problems for about eight months. Liquid
is not starting to become darker and small build up of
stuff in chiller main filter. It is time to change liquid and
filters. Main filter is in the water tank and is spec'd
at about 50u. External toilet paper style filter is spec'd
at 2u. So both get changed at the same time.

The Haskris unit specifically says to not use automotive
antifreeze since it will deteriorate the BUNA N material in
the chiller. Some anti freeze contains ethylene glycol.
So I'm puzzled by the successful use of distilled water and
EG. Perhaps they meant to say not to use 100% antifreeze
rather than a diluted mix.

The other factor is that the SEM came with basically transparent
water hoses. This is not good since the light gets into them
and advances the algae. So this upcoming liquid and filter
replacement will include replacing the hoses with opaque ones.

Overall, there are three aspects to be concerned about:

1. chiller guts and pump
2. hoses
3. SEM items that get chilled water (TMP, coils, etc.)

I don't think that there is a single simple answer to this
problem since SEMs are different and chillers are different.

gary g.



At 02:04 PM 2/17/2006, you wrote:



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From: dljones-at-bestweb.net
Date: Fri, 17 Feb 2006 20:43:14 -0600
Subject: [Microscopy] Re: Microscope cooling lines

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

Gary,

Just a couple of points I'd like to make w.r.t. your post.

I would not recommend using ethylene glycol as it is toxic. Propylene
glycol is non-toxic.

The problem with using "antifreeze" mixtures not intended for use in these
kinds of systems is that they do not usually contain the proper
additives. Glycol solutions that are not HVAC grade will deteriorate over
time through a type of polymerization that will plug things up and render
the system inoperable. The resulting deposits thus formed are very inert
and to my knowledge no one has ever found a way to clean them so you
basically have to replace the chiller system.

HVAC grade polypropylene contains additives to avoid that problem plus
inhibitors that stop corrosion of most commercially available materials in
piping. That also includes seal materials, but I'm not sure about
specifically N-buena seals. I'd have to look that up, but I would think it
also compatible being such a commonly used seal material.

Biofouling is quite common in closed loop systems. Using a biocide is
usually used in these systems to eliminate this problem.

The original poster to this thread likely is in a location where they have
a professional water treatment company taking care of large HVAC systems.
Perhaps they should talk with the representative of that company and find
out what is being done for chemical treatment of chilled water systems
there. They may be able to just get some of the proper chemicals that
likely exist on-site already.

I would also like to point out, there is really no reason to go through
the expense of using a glycol based system unless there is danger of
freezing the coolant for some reason. There are numerous other water
treatments that are much less expensive and work very well to keep a
closed loop system running well. If there is little to no make up water
needed for the closed system, once set-up properly, there is little more
to do other than enjoy a clean running system...

dj

On Fri, 17 Feb 2006, gary-at-gaugler.com wrote:

}
}
}
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
} To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
} On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} ----------------------------------------------------------------------------
}
} I had the same sludge problem about 1.5 years ago in
} a new Haskris chiller. Haskris recommends using only
} distilled water. That lasted about three months and
} the slime appeared. It was a combination of algae and
} small particles. Dumped the water and replaced with
} new distilled water and Skasol to flush. Then, new
} distilled water and one half liter of Hexid A4 from
} Applied Thermal Control Ltd. UK as supplied by SEM
} service tech. Chiller seized after about three months.
} Post mortem indicated that the impeller blades failed.
} Most likely due to misalignment of motor and pump.
}
} Haskris replaced motor and pump assembly. Fluid was
} drained and replaced with distilled water and ethylene
} glycol (.5G to 4.5G distilled water). Filters were
} changed and no problems for about eight months. Liquid
} is not starting to become darker and small build up of
} stuff in chiller main filter. It is time to change liquid and
} filters. Main filter is in the water tank and is spec'd
} at about 50u. External toilet paper style filter is spec'd
} at 2u. So both get changed at the same time.
}
} The Haskris unit specifically says to not use automotive
} antifreeze since it will deteriorate the BUNA N material in
} the chiller. Some anti freeze contains ethylene glycol.
} So I'm puzzled by the successful use of distilled water and
} EG. Perhaps they meant to say not to use 100% antifreeze
} rather than a diluted mix.
}
} The other factor is that the SEM came with basically transparent
} water hoses. This is not good since the light gets into them
} and advances the algae. So this upcoming liquid and filter
} replacement will include replacing the hoses with opaque ones.
}
} Overall, there are three aspects to be concerned about:
}
} 1. chiller guts and pump
} 2. hoses
} 3. SEM items that get chilled water (TMP, coils, etc.)
}
} I don't think that there is a single simple answer to this
} problem since SEMs are different and chillers are different.
}
} gary g.
}
}
}
} At 02:04 PM 2/17/2006, you wrote:
}
}
}
} } ----------------------------------------------------------------------------
} } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
} } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
} } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} } ----------------------------------------------------------------------------
} }
} } My service engineer recommends neither tap nor distilled water, but
} } rather bottled spring water. Has anyone yet mentioned the possibility
} } that green sludge in the filter might be algae growing in the cooling
} } water or the cooling unit?
} } --Jan Factor
} }
} } ---------------------------------------
} } Jan Robert Factor, Ph.D.
} } Professor of Biology
} } ---------------------------------------
} } Natural Sciences
} } Purchase College, State University of New York
} } 735 Anderson Hill Rd.
} } Purchase, NY 10577
} } USA
} } ---------------------------------------
} } Office Tel: 914-251-6659
} } Office Fax: 914-251-6635
} } E-mail: jfactor-at-ns.purchase.edu
} } or- jan.factor-at-purchase.edu
} } ---------------------------------------
} }
} }
} }
} } ==============================Original Headers==============================
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} 15, 20 -- Subject: Re: [Microscopy] Re: Microscope cooling lines
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From: gary-at-gaugler.com
Date: Fri, 17 Feb 2006 20:51:47 -0600
Subject: [Microscopy] Re: Microscope cooling lines

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


Thanks for the input.

If EG is toxic and PG is not, that is a factor. However,
I do not care if one or the other is toxic. I do not swim
in the 5 gallons of fluid. So, between the two glycols,
which is best for SEM? I do not know. What do you think?

So there are toxic issues, corrosive issues, etc., etc.
Well, then just dump the SEM, eh? No. I do not think so.
One must work with the materials at hand. So, what do you
think are the best materials for chiller fluid?

gary g.



At 06:43 PM 2/17/2006, you wrote:
} Gary,
}
} Just a couple of points I'd like to make w.r.t. your post.
}
} I would not recommend using ethylene glycol as it is toxic.
} Propylene glycol is non-toxic.
}
} The problem with using "antifreeze" mixtures not intended for use in
} these kinds of systems is that they do not usually contain the
} proper additives. Glycol solutions that are not HVAC grade will
} deteriorate over time through a type of polymerization that will
} plug things up and render the system inoperable. The resulting
} deposits thus formed are very inert and to my knowledge no one has
} ever found a way to clean them so you basically have to replace the
} chiller system.
}
} HVAC grade polypropylene contains additives to avoid that problem
} plus inhibitors that stop corrosion of most commercially available
} materials in piping. That also includes seal materials, but I'm not
} sure about specifically N-buena seals. I'd have to look that up, but
} I would think it also compatible being such a commonly used seal material.
}
} Biofouling is quite common in closed loop systems. Using a biocide
} is usually used in these systems to eliminate this problem.
}
} The original poster to this thread likely is in a location where
} they have a professional water treatment company taking care of
} large HVAC systems. Perhaps they should talk with the representative
} of that company and find out what is being done for chemical
} treatment of chilled water systems there. They may be able to just
} get some of the proper chemicals that likely exist on-site already.
}
} I would also like to point out, there is really no reason to go
} through the expense of using a glycol based system unless there is
} danger of freezing the coolant for some reason. There are numerous
} other water treatments that are much less expensive and work very
} well to keep a closed loop system running well. If there is little
} to no make up water needed for the closed system, once set-up
} properly, there is little more to do other than enjoy a clean running system...
}
} dj
}
} On Fri, 17 Feb 2006, gary-at-gaugler.com wrote:
}
} }
} }
} }
} } ----------------------------------------------------------------------------
} } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America


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From: tonygr-at-MIT.EDU
Date: Sat, 18 Feb 2006 07:54:12 -0600
Subject: [Microscopy] TEM film electrostatic discharging

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This is a perennial problem, I know, but does anyone have any effective "pet" solutions to the problem of charging and subsequent electrostatic arcing of TEM film in the extremely dry air we get in New England typically in the winter, and some other people get at other seasons of the year?

The discharges can occur at almost any stage of the handling, from getting the film out of the vendor's packing, loading and unloading in the microscope carriers and loading in the developing racks (we use Lucite ones). We have considered a humidifier in the darkroom, but that would involve blocking off the ventilation (we have make-up air positively blown into to room and exhaust actively sucked out) to prevent our moisture from being simply blown away.

Any hints would be welcome, but switching to digital imaging is one that is not going to happen.

Tony.

*************************************
Anthony J. Garratt-Reed M.A. D.Phil.
MIT Room #13-1027
77 Massachusetts Avenue
Cambridge, Massachusetts 02139-4307
USA

Tel: (617) 253-4622
Fax: (617) 258-6478
*************************************


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From: dsherman-at-purdue.edu
Date: Sat, 18 Feb 2006 10:40:08 -0600
Subject: [Microscopy] Cooling lines-responses

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Listers,

Below are responses (abbreviated) to my question about which to use, copper
or PVC, to replace water lines running from cooling units to microscopes. I
will run any suggestions by my service engineers just to make sure there are
no concerns regarding specific equipment. I do know that water pH is
important and that may play a role in determining what additives to use:


I am a service engineer who works on electron microscopes for the last
17 years and my point of view: I would go with copper this way no matter
what may or may not happen to the lines (cleaning out - whatever) you
have no worries. (Ray Spengler)

I do not think that there will be a problem using copper or PVC. But you
chould check with the eq maker to see if they have any issues.
We use Copper and have not had any problems. We do get build up over time
due to the hoses "rotting" over time and general scale buildup. (Karl Weiss)

USE PVC and make the final connection to the equipment with two coils of
rubber hose for vibration control. ( Fran Laabs)

For our new IMAGE building, I specified PVC piping for the closed
circ loop between the water chillers and scopes. We still notice a
greenish sludge building up in the water filters (takes about 6-8
months to become significant) but I am certain that this is coming
from the EM (copper cooling coils and iron
connections---} electrolytic reaction). The EM service people told us
that if we ever used acid to clean the lines that they would no
longer warranty the microscope. The PVC lines are perfectly clean. (J.
Bozzola)

Another material you may wish to consider is called PEX, which is a
cross-linked polyethylene. I don't know too much about its characteristics,
except that it's very smooth inside, which should retard crud accumulation
and it's more opaque than white pvc. It may be worth checking out. (P.
Grover)

As an FYI, if you have copper based and iron based materials mixed in the
same system, the iron will corrode preferentially through galvanic
coupling, not the copper. In fact, the iron becomes a sacrificial anode
protecting the
copper from corrosion. Any time you have those two materials in the same
system, you should have dielectric couplings between the two or you will
actively corrode the ferrous based material.
Regarding acid cleaning, you should know what your deposits are in your
piping before deciding how to clean them. As an FYI, copper will generally
corrode at pH's below about 6.3 or so. There are low pH cleaners that can be
used with copper, but they contain corrosion inhibitors. (D. Jones)

If you intend to clean the lines with acid, I suggest PVC, since Cu
can be etched at low pH. In addition to floating some dichlorophene
for algal control, we add a corrosion inhibitor. We have been using a
Mo-based formula, which was available from Aqua Labs on the East coast
and from Skasol on the West coast, so find a distributor in your area.
I think that either Aqua or Skasol would be able to give you that info.(B.
Tivol)

Along these lines, I noticed a lot more copper leaching into the
lines when I used deionized water than regular or filtered tap water.
Secondly, Why hasn't anyone suggested using antifreeze? Antifreeze
intended for cars should have corrosion inhibitors and should be
suitable for this purpose.
I have still gotten green particles regardless of whether antifreeze
was used or not and have regularly made it a practice to clean the
filters in the lines once a semester. (Jerry Calvin)

I have used the ethylene/glycol/water mix through
clear PVC piping between my circulator and TEM and
vacuum coating unit for six years now. There is a long
run of the piping in daylight. Absolutely no algae
Problem. Use 50/50 ethylene glycol*/water and PVC pipes and you
will never have a problem again.
Make sure that the chiller/recirculator manufacturer
OKs the use of ethylene glycol but I wouldn't
anticipate any problem. Even 20/80 EG/Water will work
well. (Ted Dunn)


Debby

Debby Sherman, Manager Phone: 765-494-6666
Life Science Microscopy Facility FAX: 765-494-5896
Purdue University E-mail: dsherman-at-purdue.edu
S-052 Whistler Building
170 S. University Street
West Lafayette, IN 47907
http://www3.agriculture.purdue.edu/microscopy



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From: soleimanij-at-tbzmed.ac.ir
Date: Sat, 18 Feb 2006 16:31:57 -0600
Subject: [Microscopy] viaWWW: scale bar

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Email: soleimanij-at-tbzmed.ac.ir
Name: Jafar Soleimani Rad

Organization: University

Title-Subject: [Filtered] scale bar

Question: Hi
we are using A LEO 906 TEM, scale bar is not registered in the
microfilms. We are having problem to calculating the real sizes. any
help in this matter is appreciated.

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From: rpowell-at-nanoprobes.com
Date: Sat, 18 Feb 2006 16:32:54 -0600
Subject: [Microscopy] viaWWW: NTA-nanogold for His tagged protein in EM?

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Email: rpowell-at-nanoprobes.com
Name: Rick Powell

Organization: Nanoprobes, Incorporated

Title-Subject: [Filtered] Re: [Microscopy]:
NTA-nanogold for His tagged protein in EM?

Question: [Commercial disclaimer - I work for
Nanoprobes, and we make NTA-Ni(II)-NanogoldĘ]

Hello Milton:

Here are a few recent references for EM using
NTA-Ni(II)-Nanogold, with links to articles on
our online newsletter that describe them:

(1) Wolfe, C. L.; Warrington, J. A.; Treadwell,
L., and Norcum, M. T.: A three-dimensional
working model of the multienzyme complex of
aminoacyl-tRNA synthetases based on electron
microscopic placements of tRNA and proteins. J.
Biol. Chem., 280, 38870-38878 (2005).

Newsletter article:
http://www.nanoprobes.com/Vol6_Iss12.html#3

(2) Bumba, L.; Tichy, M.; Dobakova, M.; Komenda,
J., and Vacha, F.: Localization of the PsbH
subunit in photosystem II from the Synechocystis
6803 using the His-tagged NińNTA Nanogold
labeling. J. Struct. Biol., 152, 28-35 (2005).

Newsletter article:
http://www.nanoprobes.com/Vol6_Iss10.html#1

There are two more recent references in which the
reagent was used to label polyhistidine-tagged
components of viral capsids:

(3) Chatterji, A.; Ochoa, W. F.; Ueno, T.; Lin
T., and Johnson, J. E.: A virus-based nanoblock
with tunable electrostatic properties. Nano
Lett., 5, 597-602 (2005).

Newsletter article:
http://www.nanoprobes.com/Vol6_Iss5.html#1

(4) Collins, R. F.; Frye, S. A.; Balasingham, S.;
Ford, R. C.; Tonjum, T., and Derrick, J. P.:
Interaction with type IV pili induces structural
changes in the bacterial outer membrane secretin
PilQ. J. Biol. Chem., 280, 18923-18930 (2005).

Newsletter article:
http://www.nanoprobes.com/Vol6_Iss6.html#6

Details about the reagent are available on our web site:

Catalog and general info:
http://www.nanoprobes.com/NTAgold.html

Product info and instructions:
http://www.nanoprobes.com/Inf2080.html

We have found that the complexes of
NTA-Ni(II)-Nanogold bound to polyhis-tagged
proteins hold together well during chromtographic
separations (gel filtration), implying that they
should be injectable.

Hope some of these are helpful,

Rick Powell
Nanoprobes, Inc.
www.nanoprobes.com


At 05:09 PM 2/17/2006, you wrote:
Question: Has anynone had success using NTA-nanogold to disclose the
location of His-tagged proteins by electron microscopy? I would like
to attach the nanogold this way before injecting the protein into a
cell. An alternative would be to apply gold labelled primary
anti-His after fixation and permeabilization.

Thanks

Milton Charlton
University of Toronto

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From: grmitch-at-netzero.com
Date: Sun, 19 Feb 2006 20:30:43 -0600
Subject: [Microscopy] AskAMicroscopist: teaching a unit on microscopic life to 5th

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Email: grmitch-at-netzero.com
Name: Linda Mitchell

Organization: BPS Environmental Center

Education: K-8 Grade Grammar School

Location: Birmingham, Michigan USA

Title: Microorganisms!

Question: I will be teaching a unit on microscopic life to 5th graders next month, March (quite a cold month here in Michigan). My question for you:
What is the best way to keep my microorganisms alive throughout the program? We obtain our samples directly from the 10 acres of the property here at the nature center. One of the samples is from our pond area and the second from the swamp woods area. We have no difficulty in finding a healthy sample, yet the problem that we often encounter is keeping them alive. We have supplies - lab pans, microscopes, even an aquarium that is our "indoor pond" when the water ices over. Do you have any feeding, climate tips that would help in these areas? This is a program that is enjoyed by both the staff and students - they are so amazed by what they find. Thanks for your input.

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From: Laurine.Ottmar-at-millenniumchem.com
Date: Mon, 20 Feb 2006 06:02:31 -0600
Subject: [Microscopy] Employment Opportunity

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Microscopist

Millennium Chemicals (a Lyondell Company), the world's second largest
producer of Titanium Dioxide, is seeking a microscopist with experience in
the areas of Scanning Electron Microscopy (SEM), Transmission Electron
Microscopy (TEM), Atomic Force Microscopy (AFM) and X-ray Diffraction (XRD)
to support R&D, Commercial and Manufacturing activities at its Baltimore
Research Center.

EDUCATIONAL REQUIREMENTS

Minimum M.S. in chemistry, mineralogy, material science or similar field,
with 7-10 years experience in an industrial R&D environment preferred.

DESCRIPTION:

The primary responsibility of the position is the application of advanced
microscopic techniques to investigate properties, mineralogy, phase
distribution, morphology and structure/function relationships of pigmentary
and catalytic TiO2 particles, catalysts and polymers. In addition to
conducting research and and support activities, the candidate will be
responsible for oversight and maintenance of a state-of-the-art microscopy
laboratory that includes:

*Olympus SZX7 stereoscope
*Olympus BX51 Optical Light Microscope
*Amray 1930 SEM with EDS (scheduled for replacement in 2006)
*Jeol 2000 FX2 with EDS and STEM
*Veeco Nanoscope 3A Dimension 3100 AFM
*Panalytical X'pert Pro XRD
*RMC PT-X Ultramicrotome with CR-X Cryosectioning system
*Gatan Model 691 Precision Ion Polishing System
*SPE Plasma Prep II Plasma Etcher/Asher
*Denton sputterer/coater

SALARY

Salary is commensurate with education and experience. Millennium Chemicals
(A Lyondell Company) offers a competitive benefits package including
relocation.

For more information and to submit a resume online, please visit our
website:

http://www.millenniumchem.com/Careers/Current+Opportunities/

Millennium Chemicals (a Lyondell Company) is an Equal Opportunity Employer
M/F/D/V






Lyondell Chemical Company

The information contained in this e-mail message and any attachments may be confidential. It is intended only for the use of the individual or entities named above. If the reader of this message is not the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify us immediately by e-mail at the originating address.


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From: richard.beanland-at-bookham.com
Date: Mon, 20 Feb 2006 10:38:12 -0600
Subject: [Microscopy] Re: TEM film electrostatic discharging

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

Tony

one thing that occurs to me is that you will probably have vinyl
flooring in your darkroom. This may be a major source of static and it
may be possible to get some advice on low static flooring. I know we
had some installed in one of our microscope cubicles and it didn't seem
to be tremendously expensive at the time.

If you think that the floor is part of the problem then it might be
worth checking whether the soles of your shoes are man made (create
static) or leather.

If you want to go for a tried and tested electronics industry route
look for ionisers (rather than de-ionisers) I did a simple Google
search on ioniser and static and came up with a few using
keywords "ionisers static"
such as http://www.buystatic.com/static_eliminators.htm?ioniser.htm

I was hoping that you could get away with a domestic ioniser - the sort
they sell to simulate sea or mountain air and supposedly gets rid of
headaches. But apparently they aren't much good at eliminating static
whereas the industrial ones are.

Good luck

Malcolm

Malcolm Haswell
e.m. unit
School of Health, Natural and Social Sciences
Fleming Building
University of Sunderland
Tyne & Wear
SR1 3SD
UK

e-mail: malcolm.haswell-at-sunderland.ac.uk

----- Original Message -----
X-from: tonygr-at-MIT.EDU

I have a friend who seems particularly prone to this even when the
humidity is not very low - she can produce amazing patterns of sparks
and spirals on a negative simply by waving her hands over them!

The simple way of getting rid of electrostatic problems in a
semiconductor company like mine is to 'borrow' an earth strap and mat
from the test area. It solved the problem completely. I guess you may
have to buy them..

Good luck

Richard

________________________________________
Richard Beanland
Analytical Services
Bookham Inc
Caswell
Towcester
Northants
NN12 8EQ
United Kingdom
Tel. +44 1327 356362
Fax. +44 1327 356775
http://www.bookham.com
________________________________________

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}
} This is a perennial problem, I know, but does anyone have any
} effective "pet" solutions to the problem of charging and
} subsequent electrostatic arcing of TEM film in the extremely dry
} air we get in New England typically in the winter, and some other
} people get at other seasons of the year?
}
} The discharges can occur at almost any stage of the handling, from
} getting the film out of the vendor's packing, loading and
} unloading in the microscope carriers and loading in the developing
} racks (we use Lucite ones). We have considered a humidifier in
} the darkroom, but that would involve blocking off the ventilation
} (we have make-up air positively blown into to room and exhaust
} actively sucked out) to prevent our moisture from being simply
} blown away.
}
} Any hints would be welcome, but switching to digital imaging is
} one that is not going to happen.
}
} Tony.
}
} *************************************
} Anthony J. Garratt-Reed M.A. D.Phil.
} MIT Room #13-1027
} 77 Massachusetts Avenue
} Cambridge, Massachusetts 02139-4307
} USA
}
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From: TindallR-at-missouri.edu
Date: Mon, 20 Feb 2006 13:19:58 -0600
Subject: [Microscopy] TEM: Cell culture preparation

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

Advice needed on TEM sample prep for cultured cells.

We are growing skin fibroblasts in culture and would like to examine the cells with transmission electron microscopy. To date, our results have been disappointing. We grew the cells on Thermanox coverslips, fixed for 1 hr at room temp in cacodylate-buffered 1.5% glutaraldehyde, 1.5% paraformaldehyde. After buffer washes, the cells were post-fixed with 1% osmium tetroxide, then washed, dehydrated in acetone and embedded in Epon/Araldite.

The cells look OK in general, but the membranes (both plasma membrane and internal membranes) are all rather fuzzy and indistinct. The quality of the ultrastructure is much poorer that we obtain with tissues prepared using almost identical proceures. One would think that since the cells are only a monolayer, preservation would be excellent.

If you have experience with TEM of cultured cells and have obtained good results, I would appreciate any advice you can give me to get better quality ultrastructure.

Please respond to me directly at katzm-at-health.missouri.edu

Thanks,

Martin L. Katz, Ph.D.
Professor
Ophthalmology, Pathobiology, Neurosciences, Genetics
Mason Eye Institute
University of Missouri School of Medicine
Columbia, Missouri  65212
Phone (573) 882-8480
FAX (573) 884-4100
katzm-at-health.missouri.edu
 http://www.muhealth.org/~ophthalmology/Katz.htm



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From: oshel1pe-at-cmich.edu
Date: Mon, 20 Feb 2006 13:57:20 -0600
Subject: [Microscopy] Re: TEM: Cell culture preparation

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

Randy/Martin,

You should be able to dispense with the formalin
(not "paraformaldehyde" -- that's the solid from
which fresh formalin is made). Monolayers of
cells generally do not need formalin.
Add 1% tannic acid to both the glut and osmium.
Mallinckrodt 1764; the monomeric form seems to
work better. You may only need the tannic acid in
one of the fixes, probably the glut, but putting
it in both the fixes won't hurt. You may need to
fix for 2 hours, but one hour should be enough.
Note: this is for high-resolution SEM, where any
holes in the plasma membrane are blatant, and it
works well up to mags } 100kX (and higher).
Also, you could try growing the cells on glass
coverslips, and digesting off the coverslip with
HF after fixation, but that's probably not
relevant to your problem.
Good luck.

Phil

} Advice needed on TEM sample prep for cultured cells.
}
} We are growing skin fibroblasts in culture and
} would like to examine the cells with
} transmission electron microscopy. To date, our
} results have been disappointing. We grew the
} cells on Thermanox coverslips, fixed for 1 hr at
} room temp in cacodylate-buffered 1.5%
} glutaraldehyde, 1.5% paraformaldehyde. After
} buffer washes, the cells were post-fixed with 1%
} osmium tetroxide, then washed, dehydrated in
} acetone and embedded in Epon/Araldite.
}
} The cells look OK in general, but the membranes
} (both plasma membrane and internal membranes)
} are all rather fuzzy and indistinct. The
} quality of the ultrastructure is much poorer
} that we obtain with tissues prepared using
} almost identical proceures. One would think
} that since the cells are only a monolayer,
} preservation would be excellent.
}
} If you have experience with TEM of cultured
} cells and have obtained good results, I would
} appreciate any advice you can give me to get
} better quality ultrastructure.
}
} Please respond to me directly at katzm-at-health.missouri.edu
}
} Thanks,
}
} Martin L. Katz, Ph.D.
} Professor
} Ophthalmology, Pathobiology, Neurosciences, Genetics
} Mason Eye Institute
} University of Missouri School of Medicine
} Columbia, MissouriŻ 65212
} Phone (573) 882-8480
} FAX (573) 884-4100
} katzm-at-health.missouri.edu
} Żhttp://www.muhealth.org/~ophthalmology/Katz.htm
--
Philip Oshel
Microscopy Facility Supervisor
Department of Biology
Central Michigan University
024C Brooks Hall
Mt. Pleasant, MI 48859
(989) 774-3576


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From: mcauliff-at-umdnj.edu
Date: Mon, 20 Feb 2006 14:25:41 -0600
Subject: [Microscopy] Re: TEM: Cell culture preparation

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

TindallR-at-missouri.edu wrote:

Hi Randy:

Many (and I do mean many) years ago I fixed cultures of retinal
pigment epithelial cells for TEM. I used a routine cacodylate-buffered
glut. fix, no formaldehyde although that should not matter. I did put 1%
tannic acid in the fix, both glut and osmium and it helped show the ECM
but I don' think it should be essential. I grew the cells in ordinary
culture dishes, no thermanox c'slips and embedded in Epon substitute. My
guess is that your membranes look fuzzy due to less than optimal
fixation of lipids (old glut and/or osmium?) or extraction of lipids
(too long in dehydration?). Why are you using acetone and not ethanol?
It is my understanding that acetone removes lipids faster than EtOH but
I have also heard that the converse it true. Certainly too long in
dehydration can degrade ultrastructure. Also, add 2mM Ca ions to the fix
to help stabilize the lipids.

Geoff

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From: mcauliff-at-umdnj.edu
Date: Mon, 20 Feb 2006 15:10:53 -0600
Subject: [Microscopy] spam filters or ??

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

Dear List;

I see that several people are using spam filters on this list. I
have just received e-mail from 2 people supposedly on this list asking
me to click on a link to verify that I am really sending them a non-spam
e-mail. No way am I going to click on a link from someone I don't know!
What ARE these people thinking?

Geoff

--
--
**********************************************
Geoff McAuliffe, Ph.D.
Neuroscience and Cell Biology
Robert Wood Johnson Medical School
675 Hoes Lane, Piscataway, NJ 08854
voice: (732)-235-4583; fax: -4029
mcauliff-at-umdnj.edu
**********************************************



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From: david.knecht-at-uconn.edu
Date: Mon, 20 Feb 2006 15:32:19 -0600
Subject: [Microscopy] TIRF setup

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

We recently saw a TIRF system demonstrated, and like other laser
based TIRF systems, it seems to use a small peripheral arc aperture
to allow the laser light through to part of the outside periphery of
the 1.45NA objective. Why would you not use a complete ring annulus
instead of an arc to illuminate the entire outside periphery of the
objective? If you simply put the correct sized annulus into the field
stop in the fluorescence path, would you get TIRF? Also, what is the
theoretical advantage of using a laser as opposed to an arc
illuminator. Thanks- Dave

Dr. David Knecht
Department of Molecular and Cell Biology
U-3125
91 N. Eagleville Rd.
University of Connecticut
Storrs, CT 06269
860-486-2200
860-486-4331 (fax)


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From: rcommon-at-msu.edu
Date: Mon, 20 Feb 2006 16:01:59 -0600
Subject: [Microscopy] TEM film electrostatic discharging

Contents Retrieved from Microscopy Listserver Archives
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For the floor, there is a spray that can be put on it to temporarily
stop electrostatic buildup. I'm sorry, but I can't remember the name of
it, but I have used it in the past.

For electronic use, I found that taking my shoes off solves static
discharge problems. There is enough moisture on my feet to stop any
discharges.

-Scott

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

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

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


-----Original Message-----
X-from: malcolm.haswell-at-sunderland.ac.uk
[mailto:malcolm.haswell-at-sunderland.ac.uk]
Sent: Monday, February 20, 2006 7:28 AM
To: Walck-at-SouthBayTech.com

Tony

one thing that occurs to me is that you will probably have vinyl
flooring in your darkroom. This may be a major source of static and it
may be possible to get some advice on low static flooring. I know we
had some installed in one of our microscope cubicles and it didn't seem
to be tremendously expensive at the time.

If you think that the floor is part of the problem then it might be
worth checking whether the soles of your shoes are man made (create
static) or leather.

If you want to go for a tried and tested electronics industry route
look for ionisers (rather than de-ionisers) I did a simple Google
search on ioniser and static and came up with a few using
keywords "ionisers static"
such as http://www.buystatic.com/static_eliminators.htm?ioniser.htm

I was hoping that you could get away with a domestic ioniser - the sort
they sell to simulate sea or mountain air and supposedly gets rid of
headaches. But apparently they aren't much good at eliminating static
whereas the industrial ones are.

Good luck

Malcolm

Malcolm Haswell
e.m. unit
School of Health, Natural and Social Sciences
Fleming Building
University of Sunderland
Tyne & Wear
SR1 3SD
UK

e-mail: malcolm.haswell-at-sunderland.ac.uk

----- Original Message -----
X-from: tonygr-at-MIT.EDU

Some of the solutions suggested seem to be based on the idea that the
operator is picking up the static charge. I think the problem is much more
likely to be caused by static on the film being discharged while unloading
the exposed negatives. I now unload film wearing rubber gloves and rarely
get a static discharge pattern on my film.

Ralph Common
Dept. of Physiology
Michigan State University


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From: frank.karl-at-degussa.com
Date: Tue, 21 Feb 2006 07:09:26 -0600
Subject: [Microscopy] Re: viaWWW: microtome section thickness

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

Interesting question....
As I understand it the colors seen in thin sections are caused by
destructive interference. The equation:
Wavelength = 2nt/m describes the color seen. n = refractive index of film
t is thickness (nm)
M is any integer.

If so it seems that as refractive index decreases, wavelength will also
decrease for the same thickness film.

I hope this help and thank you for giving me a chance to thumb through my
old reference books, it brought back memories....

Frank Karl
Degussa Corporation
Akron Technical Center
3500 Embassy Parkway
Suite 100
Akron, Ohio 44333


330-668-2235 Ext. 238


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From: David.Patton-at-uwe.ac.uk
Date: Tue, 21 Feb 2006 08:55:34 -0600
Subject: [Microscopy] spam filters or ??

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

I ignore these suspicious requests as well. I guess those folk don't
get much mail now.

Dave

-----Original Message-----
X-from: mcauliff-at-umdnj.edu [mailto:mcauliff-at-umdnj.edu]
Sent: 20 February 2006 21:15
To: David Patton

Dear List;

I see that several people are using spam filters on this list. I
have just received e-mail from 2 people supposedly on this list asking
me to click on a link to verify that I am really sending them a non-spam

e-mail. No way am I going to click on a link from someone I don't know!
What ARE these people thinking?

Geoff

--
--
**********************************************
Geoff McAuliffe, Ph.D.
Neuroscience and Cell Biology
Robert Wood Johnson Medical School
675 Hoes Lane, Piscataway, NJ 08854
voice: (732)-235-4583; fax: -4029
mcauliff-at-umdnj.edu
**********************************************



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From: hyi-at-emory.edu
Date: Tue, 21 Feb 2006 10:06:29 -0600
Subject: [Microscopy] Re: TEM: Cell culture preparation

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

Hi, Randy:

           I have heard other people report situation like this and I,
too, have experienced the same thing. Often time (not always) monolayer
cells showed very little membrane contrast, even though the tissue
processed same way had no problem. The problem with monolayer cells
seemed random regardless what types of cells were being processed. More
interestingly, I have not seen this problem with cell suspensions. In
the past, I tried to use freshly made OsO4 once when I had low membrane
contrast problem with monolayer cells, and that helped. But I still do
not understand why the problem only occurs in monolayer cells. I do not
think it is a reagent penetration issue, nor a problem of inadequate
processing protocol. Is it possible that some kind of coating material
used in culture makes it harder for OsO4 to react with lipid
molecules? 
Thank you.

Hong
Emory EM




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From: krueger.eugene-at-mayo.edu
Date: Tue, 21 Feb 2006 10:56:47 -0600
Subject: [Microscopy] Re: TEM: Cell culture preparation

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I process monolayers of cells frequently for TEM. The problem of low membrane contrast is due to the lipids being extracted away during dehydration and embedding. Even membranes "stabilized" with OsO4 can be extracted with the long processing times used during "standard fixations". I typically dehydrate in EtOH for 1-2 minutes per EtOH grade, with my samples dehydrated from 100% water to 100% EtOH in 15-20 minutes. I also keep the time in liquid resin to a minimum...my whole embedding protocol takes less than 3 hours. After I shortened my times considerably, my membranes started looking very nice and crisp!
Good Luck
-Eugene

Eugene W. A. Krueger
Sr. Research Technologist in Cell Biology II
Center for Basic Research in Digestive Diseases
Mayo Clinic and Foundation
(507)284-0580 (lab)
(507)284-0762 (fax)
krueger.eugene-at-mayo.edu




} Hi, Randy:
}
} I have heard other people report situation like this and I,
} too, have experienced the same thing. Often time (not always) monolayer
} cells showed very little membrane contrast, even though the tissue
} processed same way had no problem. The problem with monolayer cells
} seemed random regardless what types of cells were being processed. More
} interestingly, I have not seen this problem with cell suspensions. In
} the past, I tried to use freshly made OsO4 once when I had low membrane
} contrast problem with monolayer cells, and that helped. But I still do
} not understand why the problem only occurs in monolayer cells. I do not
} think it is a reagent penetration issue, nor a problem of inadequate
} processing protocol. Is it possible that some kind of coating material
} used in culture makes it harder for OsO4 to react with lipid
} molecules?
} Thank you.
}
} Hong
} Emory EM
}
}
}
}
} ==============================Original Headers==============================
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From: tivol-at-caltech.edu
Date: Tue, 21 Feb 2006 13:32:32 -0600
Subject: [Microscopy] Re: TEM film electrostatic discharging

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On Feb 18, 2006, at 5:54 AM, tonygr-at-MIT.EDU wrote:

} This is a perennial problem, I know, but does anyone have any
} effective "pet" solutions to the problem of charging and subsequent
} electrostatic arcing of TEM film in the extremely dry air we get in
} New England typically in the winter, and some other people get at
} other seasons of the year?
}
} The discharges can occur at almost any stage of the handling, from
} getting the film out of the vendor's packing, loading and unloading in
} the microscope carriers and loading in the developing racks (we use
} Lucite ones). We have considered a humidifier in the darkroom, but
} that would involve blocking off the ventilation (we have make-up air
} positively blown into to room and exhaust actively sucked out) to
} prevent our moisture from being simply blown away.
}
} Any hints would be welcome, but switching to digital imaging is one
} that is not going to happen.
}
Dear Tony,
When removing the film from the envelopes, keep it in a pack, so none
of the films rubs along any other, then lift the cardboard without
rubbing. If you can maintain contact with a grounded metal
surface--the water pipes are good--this will help a lot. Try not to
let your lab coat (or other clothing for that matter) to move along
your body or other items of clothing. Always separate films by bending
the top one away from the others, then lifting. Trade your lucite
racks for metal ones. These procedures reduced, but did not completely
eliminate, the problem when I was in Albany. When using LoDose film in
total darkness, I could at least see the discharges when they occurred.
Yours,
Bill Tivol, PhD
EM Scientist and Manager
Cryo-Electron 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: gwe-at-ufl.edu
Date: Tue, 21 Feb 2006 14:18:41 -0600
Subject: [Microscopy] MT-1 for free

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We have a vintage Sorvall MT-1 ultramicrotome available for the cost of
shipping. Manual included. Has no stereomicroscope with it. Great for
semithin plastic sections. All manual operation. It is headed for the
junk pile if no one adopts it.

--
Gregory W. Erdos, Ph.D,
Assistant Director, Biotechnology Program
Scientific Director, EM Core Lab
P.O. Box 118525
University of Florida
Gainesville, FL 32611
gwe-at-ufl.edu
Phone: 352-392-1295
Fax: 352-846-0251


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From: gwe-at-ufl.edu
Date: Tue, 21 Feb 2006 14:45:20 -0600
Subject: [Microscopy] Free M-1 has already been claimed

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The free MT-1 has already found a new home.

--
Gregory W. Erdos, Ph.D,
Assistant Director, Biotechnology Program
Scientific Director, EM Core Lab
P.O. Box 118525
University of Florida
Gainesville, FL 32611
gwe-at-ufl.edu
Phone: 352-392-1295
Fax: 352-846-0251


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From: krueger.eugene-at-mayo.edu
Date: Tue, 21 Feb 2006 14:47:11 -0600
Subject: [Microscopy] embedding schedule for monolayers

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I was asked to post my embedding schedule, so here it is:

When embedding monolayers (~60 - 90% confluent) in epoxy resin (I prefer Quetol 651) I follow this schedule:

graded series of EtOH, 25%, 50%, 70%, 95%, 100%, 100%, total dehydration time 15-20'
100%EtOH:Quetol 651 (1:1), ~25'
100% Quetol 651, ~50'
100% Quetol 651, ~100'
fresh 100% Quetol 651, then into 60C oven

I do my processing in a 6 or 12 well plate that sits on a shelf in the hood (NOT on a rotator), and I change to a new 6 or 12 well plate between changes of 100% EtOH. I also use minimal volumes of resin to decrease extraction of lipid components. If you use a more viscous resin, times may need to be adjusted longer.
Good Luck!

-Eugene
Eugene W. A. Krueger
Sr. Research Technologist in Cell Biology II
Center for Basic Research in Digestive Diseases
Mayo Clinic and Foundation
(507)284-0580 (lab)
(507)284-0762 (fax)
krueger.eugene-at-mayo.edu

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From: tivol-at-caltech.edu
Date: Tue, 21 Feb 2006 18:39:29 -0600
Subject: [Microscopy] Re: viaWWW: scale bar

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On Feb 18, 2006, at 2:32 PM, soleimanij-at-tbzmed.ac.ir wrote:

} we are using A LEO 906 TEM, scale bar is not registered in the
} microfilms. We are having problem to calculating the real sizes. any
} help in this matter is appreciated.
}
Dear Jafar,
I would calibrate the magnification(s) of interest either with a
cross-grating replica or, preferably, a Mag*I*Cal, both of which are
available from many supply houses. If you need sizes on prints, there
is a calibration slide that can be photographed in an enlarger, with
which you can determine the ratio of print sizes to negative sizes; I
don't know where to get this, but it must be widely available.
Yours,
Bill Tivol, PhD
EM Scientist and Manager
Cryo-Electron 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: milesd-at-us.ibm.com
Date: Tue, 21 Feb 2006 20:05:08 -0600
Subject: [Microscopy] Re: spam filters or ??

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It is MOST likely that those emails did not come from the people they look
like they came from.

Darrell




mcauliff-at-umdnj.ed
u
To
02/20/2006 04:11 Darrell Miles/Fishkill/IBM-at-IBMUS
PM cc

Subject
Please respond to [Microscopy] spam filters or ??
mcauliff-at-umdnj.ed
u











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

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


Dear List;

I see that several people are using spam filters on this list. I
have just received e-mail from 2 people supposedly on this list asking
me to click on a link to verify that I am really sending them a non-spam
e-mail. No way am I going to click on a link from someone I don't know!
What ARE these people thinking?

Geoff

--
--
**********************************************
Geoff McAuliffe, Ph.D.
Neuroscience and Cell Biology
Robert Wood Johnson Medical School
675 Hoes Lane, Piscataway, NJ 08854
voice: (732)-235-4583; fax: -4029
mcauliff-at-umdnj.edu
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From: mayas003-at-yahoo.com
Date: Tue, 21 Feb 2006 22:58:21 -0600
Subject: [Microscopy] Scanner for TEM Micrographs

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

Greetings:

I'm looking for a reliable scanner for TEM
Micrographs. For the past 4 years I used an old HP
scanner that was very good with 35mm and TEM
micrographs, unfortunate this scanner is no longer
working, now I need a scanner that can capture a
reliable image for routine everyday micrographs; in
order to shorten darkroom time. Any subjection?

Thanks in advance

Omayra Velez
Life Cell
Branchburg, NJ


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From: mcauliff-at-umdnj.edu
Date: Wed, 22 Feb 2006 09:02:53 -0600
Subject: [Microscopy] Re: Scanner for TEM Micrographs

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

Dear Omayra,

Like microscopes, it really is the more you pay the better the result (even
my kids £60 DX5 microscope has 250x magnification and a CMOS 'camera').

However, from my experience even going to £2,000 for a scanner, the result
of a negative or slide is sometimes not as good as the original if you
really start magnifying it up (although I haven't tried the latest 4000 to
5000 dpi top of the range Nikon slide scanners). This is mainly due to film
grain size optical effects that even Digital GEM doesn't seem to wholly
eliminate (it is just a software image processing system even if embedded
into the scanner hardware & optimised for the scanner). Grain size optical
effects are such that a 400ASA negative colour scan at 2,700dpi can produce
an appallingly unusable image that image processing can't really save -
garbage in garbage out (whereas a reflective scan of the 6x4" print produces
a very reasonable A4 image). Higher resolutions approaching or exceeding the
grain size of the file (e.g. 4000 dpi and above) are supposed to reduce this
problem considerably but Nikon type slide scanners are expensive and very
limited in negative size options. In practice many problems in image quality
are as much due to ease of magnifying a digital image compared to the
negative - a few clicks and you have a 'print' the size of a wall.

However the scanned slide image from lower ASA 50-100 slides at 4000dpi is
probably better on the VDU than most standard (not enlargements) prints made
from the negative/slide and it can easily go up to A3 in output to a
printer, which is probably all that really matters (and you still have the
negative in the likely event the next generation scanners will get even
better for the price). However enlargement prints from the negative may be
better than a magnified scanned image. Naturally if you want to zoom in on a
negative, it would have been better to do this on the TEM in the first place
and take another picture instead (wise after the event).

In practice it might also be that our eyes prefer a fuzzy analogue gradation
of colour rather than the very defined little squares of a pixellated image
at the same resolution. We are also good at discerning contrast. Also
remember that digital camera's often do some image processing during capture
(e.g. noise reduction, colour correction and sometimes even sharpen) so you
have to work on the image after scanning to get the same result. Top of the
range scanner software and hardware can do this 'automatically' using
calibration targets and so forth (e.g. Digital GEM, SHO, ICE and Silverfast
Ai). This scan processing can triple scan times to 10 minutes or more, but
can save far more than that on manual Photoshop type processing times
afterwards (and make a better job of it). Note that we can only discern 191
grey levels so 8-bit (256 grey levels) B&W scans be fine for most uses
except perhaps image analysis - and this really reduced image file size.

So the scanned images can look very good at A3 and on a 22" monitor, and if
your negative archive is going the way of my collection of 1950's 35mm home
slides and literally disintegrating into brown mush, anything is an
improvement. Surprisingly the twain software can also have an effect on
scanned quality in some cases (cheaper slide scanners often benefit from
using Silverfast SE over the cheaper bundled software).

To scan many large negatives at high resolution via a 5000dpi Creo type
hi-resolution flatbed scanner would need an investment of £12k to £45 and
the massive files would be hard to manage and archive without image size
reduction and compression which renders the whole procedure rather
pointless. Given the cost it would seem to be preferable to pay someone to
scan the odd few negatives you need scanned at this resolution with their
Creo. Hi-resolution drum scanners up to 1500dpi cost nearer £70k but if your
collection is something like NASA's archive that price can be well worth
paying (see their results at http://grin.hq.nasa.gov). A 2k x 2k or greater
pixel image digital camera system for a TEM costs around £20k to £70k.

You can get pretty good results from the new breed of sub-£1,000 flatbed
scanners though, particularly with large format negatives (i.e. TEM). Have a
look at the reviews of these semi-pro flatbed scanners on the net (around
£300 to 800 to buy). Below is an excellent link for the Canon 9950F that
could be used for 4x5" or 9 x 6cm film. It also compares the results with
the similar Epson 4990 Photo.

http://www.photo-i.co.uk

I use the Canon 9950F (£260) at home for 35mm negatives and slides. The
Canon's main weakness is that it's twain interface can only scan to the
frame sizes (i.e. not to my square Kodak 125 slides, where it chops on the
top and bottom of the slides to fit it to 35mm - a real pain). With
Silverfast SE [twain] also provided, you can scan any film size. However
Canon won't share the FARE dust removal technology with Silverfast so FARE
isn't supported - not a problem with B&W negatives as FARE and ICE only work
with colour - although you can scan B&W negatives in 'colour' with FARE/ICE
and convert to B&W in Photoshop - but 'results may vary'. I have upgraded to
Silverfast Ai for $115 for my Canon 9950F which adds in auto multi-slide
scans (but not FARE). Canon no longer manufacture dedicated slide scanners
as they feel this flatbed is as good.

At work I use the Epson 4990 (£300). It has a better twain interface and has
an A4 negative scan feature that can scan any smaller negatives in multiples
if they don't fit the standard frames provided. Silverfast SE also provided
supports ICE infra-red dust removal - but dust removal can reduce image
quality a little if the negative isn't that dusty. I use a standard rubber
bulb to blow dust off before scanning - air jets canisters are gone in a day
and can squirt propellant over the film. It's image quality is identical to
Canon 9950F for 35mm slides, although the Epson can only scan 8 slides at
one go to the Canon's 12. Scan time is similar for both (very slow with
FARE/ICE - about 10 minutes a 35mm slide). However both these flatbeds are
really 2,400 dpi not 4800 dpi as claimed - if you scan 35mm at 4800 dpi the
image quality is virtually identical the 2,400 dpi scan. However you can't
regularly scan full TEM at 4,800 dpi and above as the image size would be
near 1 Gb - we scan at 800 to 1,200 dpi mainly - although you can of course
scan small areas at 2,400 dpi resolution for 'enlargements'. All uses say
they are happy with the TEM scan quality.

If you keep the negative anyway, I would have thought being able to print to
A3 size is adequate for most purposes. The cheap Canon 9950F flatbed has
replaced my 2,700 dpi SCSI Scanwit 2740s dedicated slide scanner at home
(largely as it's so much faster and easier to use). I have to say the image
quality of these flatbeds is a little out of focus (or 'soft' as we call it)
at full magnification, but the careful use of USM (unsharp mask) in
Photoshop can improve this a lot. But they are fine up to A3 at least (from
a 35mm slide). Flatbed scans always need a little more Twain and post scan
tweeking than dedicated film scanners. Leave things like USM and colour
balance to Photoshop but use the twain interface to set things like
brightness in dark negatives and dust ICE/FARE removal.

These scanners can come with expensive Silverfast Ai and targets though
(sometimes as a Pro version) or you can upgrade at silverfast.com. But a lot
of this is for accurate colour (not TEM's strong point) - although
Silverfast is a powerful & complicated Twain interface.

When going from 2,700 dpi of the old slide/negative scanners up to the 4,000
dpi of modern Nikon type slide/negative scanners many users report far
better image quality, and put it down to reduced effects from grain aliasing
e.g. http://hardware.mcse.ms/message144915.html
and presumably the same will be true of TEM negatives and the latest 4,000
dpi semi-pro flatbed scanners (that can take 4"x5") as well. At these
resolutions film grain is also very apparent though, as 35mm film grain has
a lower dpi.

There's also dedicated & cheap large film scanners like the Epson F3200
going to 4x5" film but again the resolution of the F3200 is quoted at 'only'
3200dpi, plus it got a duff review :
http://www.photo-i.co.uk/Reviews/interactive/Scanners/Epson_F3200/page-1.htm

In fact it is probably a little better for image quality (prior to USM) than
the flatbeds for large B&W negatives, but you may be limited to the frame
sizes so check if you have 6x9cm negatives rather than proper ¼ plate. It
seems that it can scan any size smaller than the largest frame - but
certainly there's no A4 negative or reflective scanning.

Have a look at:
http://www.photoscientia.co.uk/Grain.htm
for discussions of grain size.

Have a look at
http://www.datamind.co.uk/merchant/resolution.htm
for some chat on pixel and image file sizes.

Hope this is of some use.

Regards

Keith


PS. This a bit large to post on the listerver, but it gave me something to
do while travelling for 4 hours on the train to & from work each day. This
follows on from similar threads about scanner a few months ago.


----- Original Message -----
X-from: {mayas003-at-yahoo.com}
To: {keith.morris-at-ucl.ac.uk}
Sent: Wednesday, February 22, 2006 5:02 AM

I put my EM negatives on a light box, mask off all other areas with
black paper (so 'extra' light does not mislead the light meter), copy
with my digital camera, then invert the image to a positive and convert
to grayscale in PhotoShop. Fast, easy and excellent results.

Geoff

mayas003-at-yahoo.com wrote:

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Neuroscience and Cell Biology
Robert Wood Johnson Medical School
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From: heckman-at-bgnet.bgsu.edu
Date: Wed, 22 Feb 2006 14:22:27 -0600
Subject: [Microscopy] Fwd: Re: TEM: Cell culture preparation

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


I am not sure why these would not be legitimate. I have gotten such requests that cited my message of some date and some subject, so they already have my e-mail address. I can easily envision an anti-spam service where a person must respond to such a challenge to get added to a white-list of senders. I have replied to a couple of those posts as instructed and have not been asked to re-register.

However, I also sent them a note that such a service is not very compatible with membership in a list server like this. Everyone who ever posts to the list will get challenged to register their e-mail address. I doubt that many will. And if I declined to register and continued to get these registration requests, I would probably add that person to my junk mailers list.

If someone halfway around the world doesn't want to accept the knowledge that gets shared via this list then that is their problem. They should not use such a challenge system on the e-mail address they use for this or other listservers.

Warren
________________________________________
X-from: milesd-at-us.ibm.com [mailto:milesd-at-us.ibm.com]
Sent: Tue 2/21/2006 8:05 PM
To: wesaia-at-iastate.edu

Hi

from previous replies on the list it seems to come down to about 3-4
basic types of film scanner:
1. Very expensive dedicated drum scanners
2. Standard dedicated film scanners like the Nikon
3. A combined flatbed and glassless scanner (Agfa used to make the Duo-
scan)
4. Standard flatbed scanner with glass.

I think that Keith has covered everything but the 3. combined scanner.
I use a Microtek Scanmaker 8700 and find it useful because there is no
glass between the optics and the film which should reduce Newton's
Rings, dust and finger marks. The new Microtek is cheaper and more
powerful than mine was. It handles true 3200 x 6400 dpi scans, 4.2 Dmax
negatives, 48 bit colour and sells for under 700 UK pounds or about 600
US dollars. I think it still comes with Silversoft scanning software
and Digital ICE (scratc