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Thanks to Pang, who had an eyepiece with a reticle. I just had to walk across campus and take it apart. The Periplan GF takes a reticle of 18.5 mm diameter inserted into the focusing eyepiece, not the fixed focus eyepiece. Remove the lower lens assembly (black) and then unscrew a small threaded ring from the “upper” end of the assembly. the reticle nests on a recessed ledge and held in place by the threaded ring.

Cynthia’s links are very useful for these older Leitz scopes.

Regards,
Glen
Glen MacDonald
Core for Communication Research
Virginia Merrill Bloedel Hearing Research Center
Cellular Morphology Core
Center on Human Development and Disability
Box 357923
University of Washington
Seattle, WA 98195-7923 USA
(206) 616-4156
glenmac-at-uw.edu

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} Hello,
} A colleague has a gray Leitz inverted microscope with Periplan GF 12.5X eyepieces, 23.2 mm outside diameter. Does anyone know whether they will take an eyepiece reticle?
} Thanks,
} Glen MacDonald
} Core for Communication Research
} Virginia Merrill Bloedel Hearing Research Center
} Cellular Morphology Core
} Center on Human Development and Disability
} Box 357923
} University of Washington
} Seattle, WA 98195-7923 USA
} (206) 616-4156
} glenmac-at-uw.edu
}
}
}
}
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Dear Chad,

There are two possibilities, based on my experience with GIFs (I've used
5 different GIFs on 5 different instruments over the past 18 years):

1. The table of defective columns and rows has been lost (or modified)
so that those genuinely defective pixels has not been correctly
tabulated;

2. The Peltier cooler on the CCD camera is not cooling (at all).

On the balance of probability, I would suggest you investigate the
second possibility first (the first would take some effort either by
accident or malicious design).

We recently had problems with a GIF model 2000 on a CM300F which was
commissioned in 1997. We had the same (or similar) error message. After
contacting Gatan, they suggested estimating the temperature of the chip
based on the dark count rate of two different exposure times (0.1 sec
and 5.1 sec I think), taking the log of that ratio and dividing by some
number (I cannot recall the exact figure- it is in my lab book back in
the office- I'll try to dig it out when I return).

After this investigation, we found the Peltier cooler wasn't cooling the
CCD chip. This was traced to a delicate copper wire that had broken that
connected the Peltier PSU to the cooler. Gatan succeeded in fixing it
(and getting the cooler to work at their offices), but did not work once
it was reinstalled (we suspect the wire had broken again in transit). It
is an open question as to whether repairing this again will fix the
problem.

Anyway, I hope this helps in some small way.

Yours, Jon

Cambridge University, UK.

On 2014-01-02 20:27, parishcm-at-ornl.gov wrote:

} The GIF on our CM200FEG is now refusing to tune its isochromatic
} surface. I tried all of the obvious easy fixes, such as making sure I
} was on vacuum, acquiring a new gain reference, acquiring a new dark
} reference, reloading the last known good GIB alignment (~3 weeks old),
} and rebooting everything that can be booted (PC, GIB, Digiscan).
}
} The error message is along the lines of "Too many defects in the
} isochromatic surface."
}
} Any thoughts on what I might try? It's conceivable that a 20-year-old
} GIF has decided to fail; any way I could diagnose if a DAC or lens has
} decided to die on us?

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Good afternoon all,

We are pleased to announce that the first meeting of the Chesapeake Microscopy and Microanalysis Society will be held on January 14th, 2014. We are very excited to have Dale Greenwalt of the Smithsonian National Museum of Natural History as our speaker that evening. If you are in the area of Bethesda MD, please come and have dinner with us. A flyer with all the information can be found on our web site (link below). We are looking for RSVPs by January 10 for our buffet at Dave and Buster's in White Flint Mall.

} http://csmicroscopy.org/wp/chesapeake-microscopy-and-microanalysis-society-inaugural-meeting/

January 14th, 2014

Where

Dave & Busters at the White Flint Mall

11301 Rockville Pike #300 • Kensington , MD

Agenda: 5:00-6:00 social hour with cash bar - 6:00-7:00 Dinner

7:00-8:00 Seminar - 8:00-8:30 Business Meeting.

Please RSVP to meetings-at-csmicroscopy.org so that we can have a headcount.

Cost: $25, cash/check at the door www.csmicroscopy.org

Happy New Year
Chris

Christine A. Brantner, PhD [C] NIH
NHLBI Electron Microscopy Core Facility
National Institutes of Health
14 Service Road West
Building 14E Room 104 MSC5570
Bethesda, MD 20892-5570

301-496-4711
chris.brantner-at-nih.gov {mailto:chris.brantner-at-nih.gov}

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Dear Chad,

The notes quite easily on calculating the CCD temperature (dated 9th Nov
2011):

Step 1. Take two gain normalized images at 0.1 sec and 5.1 sec (blanked
beam) and measure the average count rates per pixel (I_0.1 & I_5.1).

Step 2. Calculate the following quotient, q:

q = 10.037 * ln ((I_5.1 - I_0.1)/65.11424)

Where ln is the natural logarithm.

Step 3. The CCD temperature, T, in Kelvin is given by:

T = 20.0*(25 - q)

Example: I measured a difference in count rate of I_5.1 - I_0.1 = 209.9
counts per pixel, giving me a quotient of 11.748 and temperature of 265K
(-8C).

I tried this several times to get some consistency: 217.7 counts =}
-15.3C, etc.

I will try to find out exactly where these figures come from and update
you if needed.

I hope this helps.

Yours, Jon

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Nestor,

A Healthy and Prosperous New Year to you, and the entire list.

Thank you for the tremendous job you at keeping the list running smoothly,
with very minimal spam. The statistics are amazing!

Best Regards,
Darrell

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***************************************************************************************
Forwarded from "Ask a Microscopist"
Please remember that the person asking the question is likely
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Using the "reply" function in your email does *not* send your answer
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****************************************************************************************

realname - Liz Smith
Email - elizabeth.smith.ctr-at-dha.mil
ORGANIZATION - Defense Health Agency
EDUCATION - Graduate College
LOCATION - Falls Church, VA, USA
SUBJECT_OF_QUESTION - Microscopy Finding - Definition
QUESTION - To Whom It May Concern,

Microscopy Finding is a named, undefined attribute within Microbe
Identification the Federal Health Information Model. Would you be able
to help us with an authoritative definition for this term.

Respectfully, Liz Smith.

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Email: asw-at-sonnax.com
Name: Amy Wall

Organization: Sonnax Industries

Title-Subject: [Filtered] Connect Nikon Epiphot 300 to digital camera

Message: Does anyone have experience connecting a Nikon Epiphot 300 metallograph to a digital
camera? We are looking at a used Epiphot 300 and want to take pictures with a digital camera. The
info I see says its simple to connect a digital camera to a side port but I find no further info on
this "simple" connection. Thank you!

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Colleagues,
I hope everyone had a relaxing break. I am about to embark on
a fairly substantial project to characterize plant cell wall
structure with immunogold labeling and imaging in FESEM. I have a few
questions before investing in reagents.

I like the idea of using secondaries conjugated to ~1 nm gold
followed by enhancement. How well does this work in practice? For my
application I am not worried about 'too much' labeling and in fact I
would like to see as much of the epitope as is present in the sample.
I gather that there are both silver as well as gold enhancers. Anyone
compare them? How does the 'shelf life' compare between a 10 nm- and
a 1 nm- conjugate?

I will be labeling the surface of sectioned plant stems, so I
am not worried about penetration through the sample. But I do wonder
whether plant tissues / cell walls interact strangely with
enhancement regents. Any experience?

Finally, and in general, I would appreciate comments on
recent (last few years?) experiences with particular products/vendors.

Many many thanks,
And those in the northern hemisphere stay warm!

Tobias
--
_ ____ __ ____
/ \ / / \ / \ \ Tobias I. Baskin
/ / / / \ \ \ Professor
/_ / __ /__ \ \ \__ Biology Department
/ / / \ \ \ 611 N. Pleasant St.
/ / / \ \ \ University of
Massachusetts
/ / ___ / \ \__/ \ ____ Amherst, MA, 01003
www.bio.umass.edu/biology/baskin
Voice: 413 - 545 - 1533 Fax: 413 - 545 - 3243

==============================Original Headers==============================
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Email: ggt-at-phys.uni-sofia.bg
Name: Gichka Tsutsumanova

Organization: Sofia University, Faculty of Physics

Title-Subject: [Filtered] SEM Hitachi S570 documentation

Message: We have in the lab a Hitachi S570 SEM and we want to upgrade it. I am looking for the
schematic documentation of the microscope column. I need to know the exact dimensions and separation
distances in the column.

Can anyone help me to find it?

Thank you in advance.

G. Tsutsumanova

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Dear all,

I'm looking for TEM grid, 3.05 mm diameter, 200 mesh, carbon coated
(about
200 A carbon thick) absolutely without impurity of Zn and Ni because I
have
to investigate (by EDS-TEM) on synthetic particles containing either Zn
as
Ni.

I already tried the Au, Cu, plastic grids and all contain some amount
of at
least one of these elements. I verified that the grids sold as Cu-grid
contain either Ni as Zn. Do you have info on carbon coated grids with
the
characteristics I need?

I hope you can help me because I don't find a solution for this
problem. I
thank you a lot.

Elena Belluso

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

Elena BELLUSO

Associate Professor in Environmental Mineralogy

Ph.D. Mineralogy and Crystallography

Dip. di Scienze della Terra, Universita' degli Studi di Torino

Via Valperga Caluso n. 35, 10125 TORINO - ITALIA

tel: (39) 011 670 51 35 - fax: (39) 011 670 51 28

e-mail: elena.belluso-at-unito.it [1]

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

Elena BELLUSO

Associate Professor in Environmental Mineralogy

Ph.D. Mineralogy and Crystallography

Dip. di Scienze della Terra, Universita' degli Studi di Torino

Via Valperga Caluso n. 35, 10125 TORINO - ITALIA

tel: (39) 011 670 51 35 - fax: (39) 011 670 51 28

e-mail: elena.belluso-at-unito.it [2]

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

_"I've... seen things you people wouldn't believe. _

_Attack ships on fire off the shoulder of Orion. _

_I watched C-beams... glitter in the dark near the Tanhauser Gate. _

_All those... moments will be lost... in time..., _

_like... tears... in... rain." _

_Blade Runner_

Links:
------
[1] mailto:elena.belluso-at-unito.it
[2] mailto:elena.belluso-at-unito.it

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Title-Subject: [Filtered] TEM sample preparation for Yeast and Fungi.

Message: Dear Listeners,
How to prepare the yeast and fungi sample for TEM.
Is there any other resin used instead of araldite.?

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Hi Elena,

I would be surprised if the stray EDX signals were coming from your
grids. They are generally quite pure (at least at the TEM EDX level).
TEM EDX detection limits will be a few tenths of an at% and grid
impurities will be more in the ppm level. Also, the fact that you are
seeing stray signal from many different grids suggests that the grids
may not be the source. The only time i've ever seen impurities by EDX
was a silicone contaminated C film where the diffusion pump oil
backstreamed into the chamber during the film preparation. That batch
of films showed huge Si contamination.

So, where is the stray signal coming from? I would suspect that you are
seeing fluorescence from hard x-rays (bremsstrahlung) generated when the
beam hits the C2 aperture. The bremsstrahlung will cause areas away
from the sample in the sample chamber to fluoresce. This can be tested
by putting the beam through a hole in the sample (called a "hole
count"). In this case you should have no EDX signal at all. If you do
see something, it is likely caused by the bremsstrahlung generated
fluorescence.

Good luck,
Henk

At 1/9/2014 5:57 AM, elena.belluso-at-unito.it wrote:
}
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} Dear all,
}
} I'm looking for TEM grid, 3.05 mm diameter, 200 mesh, carbon coated
} (about
} 200 A carbon thick) absolutely without impurity of Zn and Ni because I
} have
} to investigate (by EDS-TEM) on synthetic particles containing either Zn
} as
} Ni.
}
} I already tried the Au, Cu, plastic grids and all contain some amount
} of at
} least one of these elements. I verified that the grids sold as Cu-grid
} contain either Ni as Zn. Do you have info on carbon coated grids with
} the
} characteristics I need?
}
} I hope you can help me because I don't find a solution for this
} problem. I
} thank you a lot.
}
} Elena Belluso
}
} ------------------------------------------------------------------------------------
}
} Elena BELLUSO
}
} Associate Professor in Environmental Mineralogy
}
} Ph.D. Mineralogy and Crystallography
}
} Dip. di Scienze della Terra, Universita' degli Studi di Torino
}
} Via Valperga Caluso n. 35, 10125 TORINO - ITALIA
}
} tel: (39) 011 670 51 35 - fax: (39) 011 670 51 28
}
} e-mail: elena.belluso-at-unito.it [1]
}
} ------------------------------------------------------------------------------------
}
} Elena BELLUSO
}
} Associate Professor in Environmental Mineralogy
}
} Ph.D. Mineralogy and Crystallography
}
} Dip. di Scienze della Terra, Universita' degli Studi di Torino
}
} Via Valperga Caluso n. 35, 10125 TORINO - ITALIA
}
} tel: (39) 011 670 51 35 - fax: (39) 011 670 51 28
}
} e-mail: elena.belluso-at-unito.it [2]
}
} ------------------------------------------------------------------------------------
}
} _"I've... seen things you people wouldn't believe. _
}
} _Attack ships on fire off the shoulder of Orion. _
}
} _I watched C-beams... glitter in the dark near the Tanhauser Gate. _
}
} _All those... moments will be lost... in time..., _
}
} _like... tears... in... rain." _
}
} _Blade Runner_
}
}
} Links:
} ------
} [1] mailto:elena.belluso-at-unito.it
} [2] mailto:elena.belluso-at-unito.it
}
} ==============================Original Headers==============================
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--
Untitled Document

Hendrik O. Colijn
*C*enter for *E*lectron *M*icroscopy & *A*nalysi*S*
1305 Kinnear Rd, Suite 100, Columbus, OH 43212

614.643.3458 Office colijn.1-at-osu.edu {mailto:colijn.1-at-osu.edu}
www.cemas.osu.edu {http://www.cemas.osu.edu}

"Time is that quality of nature which keeps things from happening all at
once. Lately it doesn't seem to be working."

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Hi Ravi,
You can use the protocol from Janice Griffith et al. in Journal of Electron Microscopy 60(3); 211-216 (2011). This is for IEM but you can also use it to prepare the sample for plastic embedding.

Groeten, Hans
The Netherlands Cancer Institute.
Amsterdam.

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Title-Subject: [Filtered] TEM sample preparation for Yeast and Fungi.

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How to prepare the yeast and fungi sample for TEM.
Is there any other resin used instead of araldite.?

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Happy new year to you all! I would appreciate your comments on the new non-radioactive Uranyl Acetate replacement Stain (a substitute for uranyl acetate). How do you like this product? What's your staining time?
I know this was discussed not long ago so email off list if you prefer.
thanks,
Beth

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Elena

You may wish to consider using SiN windows mounted on thin Si wafers.
These are available at most EM Supply houses. I have used these for
supporting small particles and have successfully done XEDS on these for
years. You will, of course, detect Si, N, and some O. I have never
seen Zn or Ni with these SiN films in instruments here at the ANL EMCenter.

Look for Si wafers ~ 100 microns thick, rather than the thicker (200
-300 micron) ones. The SiN windows range from 10-100 nm, obviously
the thinner the support window the better, but admittedly the 10nm
thick windows are fragile. Just be careful, or start with thicker
windows and work your way down.

Finally, be cognizant of which side you deposit your particles and also
which side is facing your XEDS detector, in order that you do not
get any shadowing effects due to the penumbra of the stage and/or
the chemically etched Si surface.

Cheers,

Nestor
Your Friendly Neighborhood SysOp

On 1/9/14 4:55 AM, elena.belluso-at-unito.it wrote:
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} Dear all,
}
} I'm looking for TEM grid, 3.05 mm diameter, 200 mesh, carbon coated
} (about
} 200 A carbon thick) absolutely without impurity of Zn and Ni because I
} have
} to investigate (by EDS-TEM) on synthetic particles containing either Zn
} as
} Ni.
}
} I already tried the Au, Cu, plastic grids and all contain some amount
} of at
} least one of these elements. I verified that the grids sold as Cu-grid
} contain either Ni as Zn. Do you have info on carbon coated grids with
} the
} characteristics I need?
}
} I hope you can help me because I don't find a solution for this
} problem. I
} thank you a lot.
}
} Elena Belluso
}
} ------------------------------------------------------------------------------------
}
} Elena BELLUSO
}
} Associate Professor in Environmental Mineralogy
}
} Ph.D. Mineralogy and Crystallography
}
} Dip. di Scienze della Terra, Universita' degli Studi di Torino
}
} Via Valperga Caluso n. 35, 10125 TORINO - ITALIA
}
} tel: (39) 011 670 51 35 - fax: (39) 011 670 51 28
}
} e-mail: elena.belluso-at-unito.it [1]
}
} ------------------------------------------------------------------------------------
}
} Elena BELLUSO
}
} Associate Professor in Environmental Mineralogy
}
} Ph.D. Mineralogy and Crystallography
}
} Dip. di Scienze della Terra, Universita' degli Studi di Torino
}
} Via Valperga Caluso n. 35, 10125 TORINO - ITALIA
}
} tel: (39) 011 670 51 35 - fax: (39) 011 670 51 28
}
} e-mail: elena.belluso-at-unito.it [2]
}
} ------------------------------------------------------------------------------------
}
} _"I've... seen things you people wouldn't believe. _
}
} _Attack ships on fire off the shoulder of Orion. _
}
} _I watched C-beams... glitter in the dark near the Tanhauser Gate. _
}
} _All those... moments will be lost... in time..., _
}
} _like... tears... in... rain." _
}
} _Blade Runner_
}
}
} Links:
} ------
} [1] mailto:elena.belluso-at-unito.it
} [2] mailto:elena.belluso-at-unito.it
}
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Dr. Nestor J. Zaluzec
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Tel: 530-NES-TORZ (530-637-8679) has Voice Mail
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The box said ...
"This program requires Win 95/98/NT or better..."
So I bought a Mac !

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For yeast cells, cryo-fixation works much better than conventional
chemical fixation. You can either use self-pressurized rapid freezing
method or high pressure freezing method. The advantage of self
pressurized rapid freezing is that you do not need specialized equipment
and it is very easy to do. I routinely use this method for processing
yeast cells. Following cryo-fixation, you will need to do
cryo-substitution and eventually embed yeast cells in resin.

You can find more information about self-pressurized rapid freezing in the
manuscript shown below:

Leunissen JL, Yi H (2009) Self-pressurized rapid freezing (SPRF): a novel
cryofixation method for specimen preparation in electron microscopy. J
Microsc (Oxford) 235:2535.

Hong

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Title-Subject: [Filtered] Sample preparation for TEM

Message: Hi Everybody

How can I prepare a sample of ZnO thin film deposited on glass substrate by RF Magnetron sputtering,
so i can analyze it in TEM

Is there any published method or somebody knows an easy way?

Thanks

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Organization: Histochemical Society

Title-Subject: [Filtered] MBL Immunohistochemistry and Microscopy Course

Message: Registration is still open for the 2014 Immunohistochemistry and Microscopy
Course (IHCM) at the Marine Biological Laboratory. Please alert your students,
postdocs and colleagues to this opportunity to attend one of MBL's Special
Topics course on Immunohistochemistry and Microscopy. Registration closes
on Tuesday, January 14! Course dates are March 15-20, 2014.

The Immunohistochemistry and Microscopy (IHCM) course is four full days and evenings (11 hours
daily) of lecture and laboratory sessions with experts in the field of immunohistochemistry (IHC)
and microscopy. The IHCM course
goal is to provide participants in-depth theory of and extensive hands-on
experience with immunohistochemistry (IHC) techniques as well as theory and
hands-on experience with a broad range of microscopic imaging techniques.
The course emphasizes hands-on laboratory time and small breakout
discussions with faculty and staff.

This course is appropriate for undergraduate and graduate students,
laboratory technicians, postdoctoral students, new and established
faculty/clinicians seeking to expand their techniques and knowledge of IHC and microscopy. It is
appropriate for beginning scientists and those with more advanced skills. Participants will be
grouped appropriately. Registration is limited to thirty.

Completed application forms and required supporting documentation must be
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FASEB Marc Awards and HCS Travel Awards are available to support
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Title-Subject: [Filtered] Save the Date: NESM's Annual Spring Meeting -at- Cabot Corporation!

Message: Dear Fellow Microscopists,

Remember to save the date for the New England Society for MicroscopyÂ’s annual Spring Meeting on
Tuesday, February 25th at Cabot Corporation! The meeting will consist of a buffet dinner and two
technical talks. More information to follow when registration opens up in the coming weeks. Keep up
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All,

I'd like to explore lab management software that includes calendar, user approval, account management, invoicing etc. I have found iLab, OCF, PPMS and Faces. Are their other products? I'd like to talk (off line) with those that use these products.

Disclaimer. I don't have a financial relationship with any of the vendors that distribute the products I mention in this message.

Thank you.

Owen

Owen P. Mills
Director, Applied Chemical and Morphological Analysis Laboratory ACMAL
Materials Science & Engineering
Michigan Technological University
1400 Townsend Drive
Houghton, MI 49931
PH 906-369-1875
FAX 906-487-2934
opmills-at-mtu.edu

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

The easiest and fastest way would be to deposit your coating on a #0 or #1
glass cover plate and use a slightly modified version of the small angle
cleavage technique. When you start scribing with the mini scribes, some
pieces will have a small angle and will be good specimens when you mount
them on the grid vertically. For ZnO, the transition region from equiaxed
growth to columnar is weak and you often get a jog in the cross section of
the film with the columnar structure back further from the tip. It just
means that you have to screen more samples in the microscope. Get your hands
on the MRS vol 480 book for Sample Prep IV and look at John McCaffrey and my
paper.

The Small Angle Cleavage Technique: An Update, S. D. Walck and J. P.
McCaffrey, Proceedings of the Materials Research Society, Workshop on
Specimen Preparation for Transmission Electron Microscopy of Materials IV,
eds. Ron M. Anderson and Scott D. Walck, Vol 480, Pittsburgh, (1997).

I think that we talk about ZnO results in this paper, but I'm not 100%
positive:
The Small Angle Cleavage Technique Applied to Coatings and Thin Films, S. D.
Walck and J. P. McCaffrey, Thin Solid Films, 308-309, pp. 399-405, 1997.

-
-Scott

Scott D. Walck, Ph.D.
Sr. Scientist
Bowhead Science & Technology
U.S. Army Research Laboratory
RDRL-WMM-C
4600 Deer Creek Loop
APG, MD 21005-5069
USA

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Email: rub.ahumada.lazo-at-gmail.com
Name: Rubén Ahumada-Lazo

Organization: Universidad Autónoma de Nuevo León

Title-Subject: [Filtered] Sample preparation for TEM

Message: Hi Everybody

How can I prepare a sample of ZnO thin film deposited on glass substrate by
RF Magnetron sputtering, so i can analyze it in TEM

Is there any published method or somebody knows an easy way?

Thanks

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Hello Listserver,

We have a couple of Hitachi CuteVac VR16L-K roughing pumps that need new vanes and oil seals. Unfortunately, nobody seems to sell parts for these. A couple of people have suggested that the parts are interchangeable with another manufacturer, (Alcatel?) but nobody has been able to provide specific part #s for the vanes or anything else.

Anyone rebuilt one of these before and remember which vanes and seals to use?

Thanks,
Bradford Ross

Microscopy Technician
BioImaging Facility
University of British Columbia
6270 University Blvd.
Vancouver, B.C.
Canada
V6T 1Z4

phone 604-822-6996

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Does anybody have a JEOL 12XX TEM gun lift hanging around in a parts bin somewhere? I am in need of one...

--Justin A. Kraft

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Name: Nabil Bassim

Organization: Naval Research Laboratory

Title-Subject: [Filtered] Registration Reminder: FIB SEM Workshop, Feb 27, 2014 and Johns Hopkins APL

Message: Call for Papers & Registration Reminder

7th Annual FIB SEM Workshop

Thursday, February 27, 2014

Kossiakoff Center

Johns Hopkins Applied Physics Laboratory

Laurel, MD

2014 FIB SEM Workshop will be held at the Kossiakoff Center at Johns Hopkins Applied Physics
Laboratory (APL). We will have a full day of presentations and posters by FIB users and vendors
highlighting interesting new FIB applications and the latest technology. And, as always, there will
be plenty of opportunities for informal discussion of new techniques and applications as well as
getting (re)connected with your fellow FIBers.

If you would like to present a paper or poster, please submit an abstract to the organizers.
Starting this year, we are requiring an abstract submission from all presenters (user & vendor
contributions). All FIB related topics such as new and novel application areas for FIB, advances in
FIB and FIB related instrumentation and techniques, image and data processing techniques for FIB
data are all welcome.

Â· Abstract Deadline: January 15, 2014

Abstracts should be 250 words or less in length. Email abstracts to fibsem2014-at-fibsem.net.

Abstract Notification: February 1, 2014

Â· Registration Deadline: February 3, 2014 - Register!

Â· Travel Info

Please check our website (www.fibsem.org) for additional information about the meeting. There is no
fee to attend the meeting, and breakfast and lunch will be provided! There will be a Happy Hour
following the meeting, so donÂ’t forget to sign up for it when you register.

Remember, you don't have to be a local user to attend! We look forward to seeing you at APL!

Organizers:

Nabil Bassim, nabil.bassim-at-nrl.navy.mil

Ken Livi, klivi-at-jhu.edu

Joan Hoffmann, joan.hoffmann-at-jhuapl.edu

Keana Scott, keana.scott-at-nist.gov

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Email: mmcgough-at-histochemicalsociety.org
Name: Meg McGough

Organization: The Histochemical Society

Title-Subject: [Filtered] MBL Immunohistochemistry and Microscopy Course - Application Deadline Extended

Message: The application deadline for the 2014 Immunohistochemistry and Microscopy
Course (IHCM) at the Marine Biological Laboratory has been extended until January 21. Course dates
are March 15-20, 2014.

The Immunohistochemistry and Microscopy (IHCM) course is four full days and evenings (11 hours
daily) of lecture and laboratory sessions with experts in the field of immunohistochemistry (IHC)
and microscopy. The IHCM course
goal is to provide participants in-depth theory of and extensive hands-on
experience with immunohistochemistry (IHC) techniques as well as theory and
hands-on experience with a broad range of microscopic imaging techniques.
The course emphasizes hands-on laboratory time and small breakout
discussions with faculty and staff.

This course is appropriate for undergraduate and graduate students,
laboratory technicians, postdoctoral students, new and established
faculty/clinicians seeking to expand their techniques and knowledge of IHC and microscopy. It is
appropriate for beginning scientists and those with more advanced skills. Participants will be
grouped appropriately. Registration is limited to thirty.

Completed application forms and required supporting documentation must be
received by January 21, 2014.

FASEB Marc Awards are available to support attendance at the course:
http://www.faseb.org/MARC-and-Professional-Development/Travel-Awards/Scientific-Meetings-and-Conferences.aspx

For further information about the course:
http://hermes.mbl.edu/education/courses/special_topics/ihcm.html

Thank you,
Meg McGough
The Histochemical Society

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Dear Elena,
Have you considered that the Zn/Ni could be coming from the TEM or sample
holder (or even the EDS detector) rather than the grid itself. When you hit
the sample with high energy elections, you generate a huge number of x-rays
of all energies in addition to backscattered electrons. If you have any
brass near your sample, you are almost guaranteed to get Cu and Zn in your
EDS spectrum from non-local fluorescence. The Ni could be coming from SS
near the sample or the TEM pole piece (do you also see Fe?).
While TEM based EDS is a lot cleaner than it once was, you can never
really be sure where the x-rays that enter your detector originate. Using
low background holders and TEM hard x-ray apertures (if available) can help.
Chapter 33.3 in Williams and Carter's TEM book details these issues and
describes precautions to help reduce system and spurious x-ray artifacts.

If you have access to an EELS detector, you can run the experiments
without worrying about fluorescence artifacts. You will only see the
element if it is under the beam.

Hope this helps.

-Ray

Disclaimer: I work for Gatan which provides EELS and EDS equipment and
software as well as low background TEM sample holders.

Ray D. Twesten
Livermore, CA

-----Original Message-----
X-from: elena.belluso-at-unito.it [mailto:elena.belluso-at-unito.it]
Sent: Thursday, January 09, 2014 3:04 AM
To: ray.twesten-at-sbcglobal.net

Dear all,

I'm looking for TEM grid, 3.05 mm diameter, 200 mesh, carbon coated (about
200 A carbon thick) absolutely without impurity of Zn and Ni because I have
to investigate (by EDS-TEM) on synthetic particles containing either Zn as
Ni.

I already tried the Au, Cu, plastic grids and all contain some amount of at
least one of these elements. I verified that the grids sold as Cu-grid
contain either Ni as Zn. Do you have info on carbon coated grids with the
characteristics I need?

I hope you can help me because I don't find a solution for this problem. I
thank you a lot.

Elena Belluso

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

Elena BELLUSO

Associate Professor in Environmental Mineralogy

Ph.D. Mineralogy and Crystallography

Dip. di Scienze della Terra, Universita' degli Studi di Torino

Via Valperga Caluso n. 35, 10125 TORINO - ITALIA

tel: (39) 011 670 51 35 - fax: (39) 011 670 51 28

e-mail: elena.belluso-at-unito.it [1]

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

Elena BELLUSO

Associate Professor in Environmental Mineralogy

Ph.D. Mineralogy and Crystallography

Dip. di Scienze della Terra, Universita' degli Studi di Torino

Via Valperga Caluso n. 35, 10125 TORINO - ITALIA

tel: (39) 011 670 51 35 - fax: (39) 011 670 51 28

e-mail: elena.belluso-at-unito.it [2]

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

_"I've... seen things you people wouldn't believe. _

_Attack ships on fire off the shoulder of Orion. _

_I watched C-beams... glitter in the dark near the Tanhauser Gate. _

_All those... moments will be lost... in time..., _

_like... tears... in... rain." _

_Blade Runner_

Links:
------
[1] mailto:elena.belluso-at-unito.it
[2] mailto:elena.belluso-at-unito.it

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Dear List

For the purpose of orientation while viewing a specimen in TEM, I naively
thought it might be useful to "color" a resin by adding electrodense grains
(e.g.ferritin) and then, after appropriate cutting, to re-embed the block in
the "colored" resin. In this way one or more side(s) of the specimen
touching the cut can be readily recognizable in the thin sections under the
TEM.

Does it make any sense what I am saying and (if yes) is there any experience
on this?

Thanks for your time

yorgos

Dr Yorgos Nikas
Athens Innovative Microscopy
Skra 36 Voula 16673 GREECE

Tel/fax +30 210 8957677
mobile +30 6945 107477
www.eikonika.net www.aim.cat
*************************************

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April 8, 2014 marks the end of support from Microsoft for Windows XP. That means no more security updates to patch newly discovered security holes. This will leave any computers still running Windows XP vulnerable to attacks if they remain connected to the internet or if infected memory media are inserted into the computer. I am sure there are a lot of machines out there running Windows XP. I am curious to hear what approaches microscopy community members are taking to deal with this, as well as what the community of microscopy related hardware vendors are doing to address this situation. Thank you.

John Mardinly, ASU

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At my place, we are putting a Win7 computer between the XPs and the corporate network. The Win7 will act as a server for the XPs. We have a few SEMs and a couple of other tools that cannot be upgraded to Win7. Unless, of course, we want to buy whole new tools!

} -----Original Message-----
} From: John.Mardinly-at-asu.edu [mailto:John.Mardinly-at-asu.edu]
} Sent: Wednesday, January 15, 2014 3:12 PM
} To: Holdford, Becky
} Subject: [Microscopy] Windows XP Support Ending April 8, 2014
}
}
}
}
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America To

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Email: sergey-at-seas.ucla.edu
Name: Sergey Prikhodko

Organization: University of California, Los Angeles

Title-Subject: [Filtered] SCSMM full-day symposium

Message: Dear all

The Southern California Society for Microscopy and Microanalysis (SCSMM)
announce their Full-Day Symposium.
The meeting will be held on Saturday, February 8th from 8:00 am at
Calit2 Auditorium at the University of California, Irvine. A lunch and
coffee breaks will be served.

Our symposium, dedicated to hybrid approaches in microscopy. Beside the
fact that it poses significant challenges for researchers in both life
and materials sciences, it continues the conversation about nanoscience,
its role, and its future. We are honored to have Prof. C. Barry Carter
from the University of Connecticut best known in the Â“microscopy worldÂ”
as the co-author of the textbook Â“Transmission Electron Microscopy: A
Textbook for Materials ScienceÂ” as our Microscopy Society of America
Invited Speaker. He will have an inspiring talk titled Â“Challenges for
TEM and TEMers: Remember the Past if You CanÂ” in which he links the
history of TEM to the current status of the field and its future. Along
with Prof. Carter, our invited speakers Prof. Suneel Kodambaka (UCLA),
Prof. Dorit Hanein (Sanford-Burnham MRI), Prof. Daniel R. Mumm (UCI) and
Dr. Yi-Wei Chang (Caltech) will make our symposium a remarkable event.

In order to register, please RSVP by email at micromark-at-juno.com.
Respond no later than 5:00 p.m. Friday, January 31st, 2014 (indicate
your special needs by this due).

The cost is $25 for professionals and $10 for students. Membership dues
can be paid at the door on February 8th, 2014.

Nonmembers are welcome to attend.

Please pass along to those you feel would be interested.

We look forward to seeing you at the meeting!

http://www.scsmm.org/

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We are keeping our XP machine unplugged from the network, and I made
sure to turn off "autorun" for all media (CD/DVD and USB) so infected
media can't take over your machine automatically (unless there's a new
way to get Windows to automatically execute files, which wouldn't
surprise me).

Cheers,

Nick Botto
Microprobe Specialist
UC Davis Earth and Planetary Sciences Microprobe Lab
Website: http://microprobe.geology.ucdavis.edu/index.html
Calendar: http://microprobe.geology.ucdavis.edu/calendar/probe.htm
ph. 530-752-6582

On 1/15/14, 1:21 PM, John.Mardinly-at-asu.edu wrote:
} ----------------------------------------------------------------------------
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} To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
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} April 8, 2014 marks the end of support from Microsoft for Windows XP. That means no more security updates to patch newly discovered security holes. This will leave any computers still running Windows XP vulnerable to attacks if they remain connected to the internet or if infected memory media are inserted into the computer. I am sure there are a lot of machines out there running Windows XP. I am curious to hear what approaches microscopy community members are taking to deal with this, as well as what the community of microscopy related hardware vendors are doing to address this situation. Thank you.
}
} John Mardinly, ASU
}
}
}
} ==============================Original Headers==============================
} 4, 35 -- From John.Mardinly-at-asu.edu Wed Jan 15 15:12:14 2014
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} 4, 35 -- To: "Microscopy-at-Microscopy.com" {Microscopy-at-Microscopy.com}
} 4, 35 -- Subject: Windows XP Support Ending April 8, 2014
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As an addendum to this, Microsoft announced on January 15th that it would continue to provide updates for security products until July 14, 2015. So no more system updates, but at least they'll continue to update Security Essentials and other MS antivirus signatures for machines running XP.

http://thenextweb.com/microsoft/2014/01/15/microsoft-extends-updates-windows-xp-security-products-july-14-2015

--
Adam Schuetze, D.Tech, B.Eng
Laboratory Engineer, Advanced Microscopy Facility
Bob Wright A015, University of Victoria, Canada
adamschu-at-uvic.ca
Lab: 250.853.3968
http://www.stehm.uvic.ca

-----Original Message-----
X-from: nwbotto-at-ucdavis.edu [mailto:nwbotto-at-ucdavis.edu]
Sent: Thursday, January 16, 2014 10:28 AM
To: Adam Schuetze

We are keeping our XP machine unplugged from the network, and I made sure to turn off "autorun" for all media (CD/DVD and USB) so infected media can't take over your machine automatically (unless there's a new way to get Windows to automatically execute files, which wouldn't surprise me).

Cheers,

Nick Botto
Microprobe Specialist
UC Davis Earth and Planetary Sciences Microprobe Lab
Website: http://microprobe.geology.ucdavis.edu/index.html
Calendar: http://microprobe.geology.ucdavis.edu/calendar/probe.htm
ph. 530-752-6582

On 1/15/14, 1:21 PM, John.Mardinly-at-asu.edu wrote:
} ----------------------------------------------------------------------
} ------ The Microscopy ListServer -- CoSponsor: The Microscopy Society
} of America To Subscribe/Unsubscribe --
} http://www.microscopy.com/MicroscopyListserver
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} April 8, 2014 marks the end of support from Microsoft for Windows XP. That means no more security updates to patch newly discovered security holes. This will leave any computers still running Windows XP vulnerable to attacks if they remain connected to the internet or if infected memory media are inserted into the computer. I am sure there are a lot of machines out there running Windows XP. I am curious to hear what approaches microscopy community members are taking to deal with this, as well as what the community of microscopy related hardware vendors are doing to address this situation. Thank you.
}
} John Mardinly, ASU
}
}
}
} ==============================Original
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5, 23 -- Message-ID: {52D82143.9030301-at-ucdavis.edu} 5, 23 -- Date: Thu, 16 Jan 2014 10:13:23 -0800 5, 23 -- From: Nick Botto {nwbotto-at-ucdavis.edu} 5, 23 -- User-Agent: Mozilla/5.0 (Macintosh; Intel Mac OS X 10.9; rv:24.0) Gecko/20100101 Thunderbird/24.2.0 5, 23 -- MIME-Version: 1.0 5, 23 -- To: microscopy-at-microscopy.com 5, 23 -- Subject: Re: [Microscopy] Windows XP Support Ending April 8, 2014 5, 23 -- References: {201401152121.s0FLLWhR023627-at-ns.microscopy.com}
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==============================Original Headers==============================
14, 31 -- From adamschu-at-uvic.ca Thu Jan 16 13:31:21 2014
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Email: jcsmtf-at-mail.missouri.edu
Name: Josh Schorp

Organization: University of Missouri

Title-Subject: [Filtered] CM10/12 heat production

Message: Does anyone know what the heat load that the CM10 and CM12 generate?

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Josh,

I've got the heat load specs for the CM-10 - just have to dig them out.
I'll send you a pdf.
Not for the CM-12, but they won't be too different.

Phil

} Email: jcsmtf-at-mail.missouri.edu
} Name: Josh Schorp
}
} Organization: University of Missouri
}
} Title-Subject: [Filtered] CM10/12 heat production
}
} Message: Does anyone know what the heat load that the CM10 and CM12 generate?
}
--
Philip Oshel
Microscopy Facility Supervisor
Biology Department
024C Brooks Hall
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576

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How stain the roots with mycorrhiza for arbuscules before maunting in EPON resin for TEM microscopy?

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Dear Listers,

we find strange bubbles when embedding Bacillus subtilis bacteria in LR
white resin. The bubbles prevent the resin from polymerising, does
anybody have an idea how and why this could happen?

http://chimerism.medunigraz.at/Pictures/pioloform01.php
http://chimerism.medunigraz.at/Pictures/pioloform02.php

Many thanks,
Stefan

==============================Original Headers==============================
5, 30 -- From stefan.wernitznig-at-medunigraz.at Tue Jan 21 09:30:56 2014
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5, 30 -- Date: Tue, 21 Jan 2014 16:30:49 +0100
5, 30 -- From: Stefan Wernitznig {stefan.wernitznig-at-medunigraz.at}
5, 30 -- User-Agent: Mozilla/5.0 (Windows NT 5.1; rv:17.0) Gecko/20130801 Thunderbird/17.0.8
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Dear Stefan

These bubbles seem to be trapped air in the resin. Put the specimens
in the final resin (in the embedding capsules) and left them for an
hour. The air bubbles will come to the surface and you can easily
break them with a needle. Then put the capsules to the oven for
polymerisation.

Panos

Quoting stefan.wernitznig-at-medunigraz.at:

} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
} To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
} On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} ----------------------------------------------------------------------------
}
} Dear Listers,
}
} we find strange bubbles when embedding Bacillus subtilis bacteria in LR
} white resin. The bubbles prevent the resin from polymerising, does
} anybody have an idea how and why this could happen?
}
} http://chimerism.medunigraz.at/Pictures/pioloform01.php
} http://chimerism.medunigraz.at/Pictures/pioloform02.php
}
} Many thanks,
} Stefan
}
}
} ==============================Original Headers==============================
} 5, 30 -- From stefan.wernitznig-at-medunigraz.at Tue Jan 21 09:30:56 2014
} 5, 30 -- Received: from si069.medunigraz.at (si082.medunigraz.at
} [193.170.105.82])
} 5, 30 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP
} id s0LFUt90018127
} 5, 30 -- for {Microscopy-at-microscopy.com} ; Tue, 21 Jan 2014 09:30:56 -0600
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} [10.200.112.128])
} 5, 30 -- by si063.medunigraz.at with ESMTP (TLS encrypted); Tue, 21
} Jan 2014 16:30:47 +0100
} 5, 30 -- Message-ID: {52DE92A9.9030803-at-medunigraz.at}
} 5, 30 -- Date: Tue, 21 Jan 2014 16:30:49 +0100
} 5, 30 -- From: Stefan Wernitznig {stefan.wernitznig-at-medunigraz.at}
} 5, 30 -- User-Agent: Mozilla/5.0 (Windows NT 5.1; rv:17.0)
} Gecko/20130801 Thunderbird/17.0.8
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} 5, 30 -- Subject: Strange bubbles in LR white resin
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} ==============================End of - Headers==============================

--
Dr. Berillis Panagiotis
Lecturer in Microscopy, image analysis and histology
Department of Ichthyology and Aquatic Environment
Agricultural school
University of Thessaly

==============================Original Headers==============================
8, 21 -- From pveril-at-apae.uth.gr Tue Jan 21 13:53:04 2014
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8, 21 -- Message-ID: {20140121215252.Horde.5K4CvlZsTmoxhFGI1v_E0w1-at-webmail.uth.gr}
8, 21 -- From: Panagiotis BERILLIS {pveril-at-apae.uth.gr}
8, 21 -- To: stefan.wernitznig-at-medunigraz.at, Microscopy-at-microscopy.com
8, 21 -- Subject: Re: [Microscopy] Strange bubbles in LR white resin
8, 21 -- References: {201401211539.s0LFds8O029163-at-ns.microscopy.com}
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Those don't look like air bubbles to me. They look to being something extracted from the bacteria. Maybe not fully dehydrated? Some non-miscible solvent still around? Air bubbles shouldn't prevent LRW from polymerizing. Does a "blank" block with no cells polymerize?

Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
phillipst-at-missouri.edu

http://biology.missouri.edu/people/?person=88
http://www.biotech.missouri.edu/mcc/

-----Original Message-----
X-from: pveril-at-apae.uth.gr [mailto:pveril-at-apae.uth.gr]
Sent: Tuesday, January 21, 2014 1:54 PM
To: Phillips, Thomas E.

Dear Stefan

These bubbles seem to be trapped air in the resin. Put the specimens in the final resin (in the embedding capsules) and left them for an hour. The air bubbles will come to the surface and you can easily break them with a needle. Then put the capsules to the oven for polymerisation.

Panos

Quoting stefan.wernitznig-at-medunigraz.at:

} ----------------------------------------------------------------------
} ------ The Microscopy ListServer -- CoSponsor: The Microscopy Society
} of America To Subscribe/Unsubscribe --
} http://www.microscopy.com/MicroscopyListserver
} On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} ----------------------------------------------------------------------
} ------
}
} Dear Listers,
}
} we find strange bubbles when embedding Bacillus subtilis bacteria in
} LR white resin. The bubbles prevent the resin from polymerising, does
} anybody have an idea how and why this could happen?
}
} http://chimerism.medunigraz.at/Pictures/pioloform01.php
} http://chimerism.medunigraz.at/Pictures/pioloform02.php
}
} Many thanks,
} Stefan
}
}
} ==============================Original
} Headers==============================
} 5, 30 -- From stefan.wernitznig-at-medunigraz.at Tue Jan 21 09:30:56 2014
} 5, 30 -- Received: from si069.medunigraz.at (si082.medunigraz.at
} [193.170.105.82])
} 5, 30 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP
} id s0LFUt90018127
} 5, 30 -- for {Microscopy-at-microscopy.com} ; Tue, 21 Jan 2014 09:30:56 -0600
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} id 51.2C.04500.EA29ED25; Tue, 21 Jan 2014 16:30:54 +0100 (CET) 5, 30
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} Tue, 21 Jan 2014 16:30:49 +0100 5, 30 -- From: Stefan Wernitznig
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} ==============================End of -
} Headers==============================

--
Dr. Berillis Panagiotis
Lecturer in Microscopy, image analysis and histology Department of Ichthyology and Aquatic Environment Agricultural school University of Thessaly

==============================Original Headers==============================
8, 21 -- From pveril-at-apae.uth.gr Tue Jan 21 13:53:04 2014 8, 21 -- Received: from kerberos.uth.gr (mx2.uth.gr [194.177.200.3])
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8, 21 -- From: Panagiotis BERILLIS {pveril-at-apae.uth.gr} 8, 21 -- To: stefan.wernitznig-at-medunigraz.at, Microscopy-at-microscopy.com 8, 21 -- Subject: Re: [Microscopy] Strange bubbles in LR white resin 8, 21 -- References: {201401211539.s0LFds8O029163-at-ns.microscopy.com}
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==============================Original Headers==============================
18, 30 -- From PhillipsT-at-missouri.edu Tue Jan 21 14:35:21 2014
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18, 30 -- To: "'microscopy-at-microscopy.com'" {microscopy-at-microscopy.com}
18, 30 -- Subject: RE: [Microscopy] Re: Strange bubbles in LR white resin
18, 30 -- Thread-Topic: [Microscopy] Re: Strange bubbles in LR white resin
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Stefan,

By chance did you happen to post fix the bacteria in osmium tetroxide? LR White does not polymerize well when osmium is used in the processing protocol.

Mary Ard
Electron Microscopy Lab Coordinator
Department of Pathology
College of Veterinary Medicine
University of Georgia
501 D.W. Brooks Drive
Athens, GA 30602
706-542-5537

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Email: paulwebsterphd-at-gmail.com
Name: Paul Webster

Organization: University of Souther California

Title-Subject: [Filtered] LR White and air bubbles

Message:
Dear Stephan,

I agree with Tom, that it looks as if something has been released from the bacteria. It could be
lipid being extracted by the resin, which is a pretty good solvent.

I am not sure that this is the reason why the resin has not polymerized. From your images, it does
look as if you have used osmium tetroxide to contrast the cells. It might also be possible that you
also used acetone to dehydrate the cells.

Residual amounts of acetone may have interfered with resin polymerization in the way you describe. A
more detailed description of your processing and embedding protocol may help diagnose the problem.

Paul Webster,
University of Southern California
Los Angeles, CA

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Panos,

What is the average size of the "particles"? You do not have a size bar in your image to get a feel for their size, which is important in any viral identification. The "particles" do not appear to be associated with the sample on the grid but laying on top of the sample. In my opinion, they look more like crystalline structures rather than viral particles. Depending on their actual size, could they possibly be phosphotungstic acid crystals?

Kind regards,
Mary

Mary Ard
Electron Microscopy Lab Coordinator
Department of Pathology
College of Veterinary Medicine
University of Georgia
501 D.W. Brooks Drive
Athens, GA 30602
706-542-5537

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Hi Panos!

Giving a picture without scale to a miscroscopist is like talking without words. We cannot make anything out if it!
I doubt these objects are embedded in the resin. One can see a hole in the section and one object seems to partly cover it. Also they don't look like they were sectioned.
Looks like artifacts for me.

Regards,
Stephane

________________________________
X-from: "pveril-at-apae.uth.gr" {pveril-at-apae.uth.gr}
To: nizets2-at-yahoo.com
Sent: Thursday, January 23, 2014 11:29 AM

Dear all

During an electron microscope examination of a Sparus aurata liver
collagen (PTA and uranyl acetate were used as stains) I came across to
these objects. They are probably virus particles. Can any one identify
them?

http://upload.users.uth.gr/files/virus.jpg

Many thanks
Panos
--
Dr. Berillis Panagiotis
Lecturer in Microscopy, image analysis and histology
Department of Ichthyology and Aquatic Environment
Agricultural school
University of Thessaly

==============================Original Headers==============================
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I agree, it does not looks like belonging to section. Some contamination after sectioning/staining. I have stained a lot with PTA, have never seen something like this.

Vladimir

Vladimir M. Dusevich, Ph.D.
Electron Microscope Lab Director
(816) 235-2072
371 School of Dentistry
650 E. 25th Street
Kansas City, MO 64108-2784

-----Original Message-----
X-from: nizets2-at-yahoo.com [mailto:nizets2-at-yahoo.com]
Sent: Thursday, January 23, 2014 8:23 AM
To: Dusevich, Vladimir

Hi Panos!

Giving a picture without scale to a miscroscopist is like talking without words. We cannot make anything out if it!
I doubt these objects are embedded in the resin. One can see a hole in the section and one object seems to partly cover it. Also they don't look like they were sectioned.
Looks like artifacts for me.

Regards,
Stephane

________________________________
X-from: "pveril-at-apae.uth.gr" {pveril-at-apae.uth.gr}
To: nizets2-at-yahoo.com
Sent: Thursday, January 23, 2014 11:29 AM

Dear all

During an electron microscope examination of a Sparus aurata liver
collagen (PTA and uranyl acetate were used as stains) I came across to
these objects. They are probably virus particles. Can any one identify
them?

http://upload.users.uth.gr/files/virus.jpg

Many thanks
Panos
--
Dr. Berillis Panagiotis
Lecturer in Microscopy, image analysis and histology
Department of Ichthyology and Aquatic Environment
Agricultural school
University of Thessaly

==============================Original Headers==============================
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25, 30 -- From: "Dusevich, Vladimir" {DusevichV-at-umkc.edu}
25, 30 -- To: "Microscopy-at-microscopy.com" {Microscopy-at-microscopy.com}
25, 30 -- Subject: RE: [Microscopy] Fw: Virus
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Depending of course on the scale, this looks a lot like a bunch of
fungus spores, similar although not identical to:

http://www.mta.ca/dmf/download/jme/092310_0016.bmp

Maybe somebody was eating a bacon-mushroom burger in the microtomy area ;-)

Jim

--

James M. Ehrman
Digital Microscopy Facility
Mount Allison University
63B York St.
Sackville, NB E4L 1G7
CANADA

phone: 506-364-2519
fax: 506-364-2505
email: jehrman-at-mta.ca
www: http://www.mta.ca/dmf

A day without fusion is like a day without sunshine.

On 23/01/2014 11:05 AM, DusevichV-at-umkc.edu wrote:
}
}
} ----------------------------------------------------------------------------
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} I agree, it does not looks like belonging to section. Some contamination after sectioning/staining. I have stained a lot with PTA, have never seen something like this.
}
} Vladimir
}
} Vladimir M. Dusevich, Ph.D.
} Electron Microscope Lab Director
} (816) 235-2072
} 371 School of Dentistry
} 650 E. 25th Street
} Kansas City, MO 64108-2784
}
} -----Original Message-----
} X-from: nizets2-at-yahoo.com [mailto:nizets2-at-yahoo.com]
} Sent: Thursday, January 23, 2014 8:23 AM
} To: Dusevich, Vladimir
} Subject: [Microscopy] Fw: Virus
}
}
}
}
} ----------------------------------------------------------------------------
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}
} Hi Panos!
}
} Giving a picture without scale to a miscroscopist is like talking without words. We cannot make anything out if it!
} I doubt these objects are embedded in the resin. One can see a hole in the section and one object seems to partly cover it. Also they don't look like they were sectioned.
} Looks like artifacts for me.
}
} Regards,
} Stephane
}
}
} ________________________________
} X-from: "pveril-at-apae.uth.gr" {pveril-at-apae.uth.gr}
} To: nizets2-at-yahoo.com
} Sent: Thursday, January 23, 2014 11:29 AM
} Subject: [Microscopy] Virus
}
}
}
}
}
} ----------------------------------------------------------------------------
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} ----------------------------------------------------------------------------
}
}
} Dear all
}
} During an electron microscope examination of a Sparus aurata liver
} collagen (PTA and uranyl acetate were used as stains) I came across to
} these objects. They are probably virus particles. Can any one identify
} them?
}
} http://upload.users.uth.gr/files/virus.jpg
}
} Many thanks
} Panos

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Ya sou Panos
Yes, scales are different, yet it is remarkable how much your particles
resemble to pollen I have a picture from a wild flower pollen that looks
very close to these particles
http://www.eikonika.net/v2/downloads/KITSTI03_resize.jpg.
Unlike in the North America, spring came earlier in Greece this year...
Best wishes
yorgos

Dr Yorgos Nikas
Athens Innovative Microscopy
Skra 36 Voula 16673 GREECE

Tel/fax +30 210 8957677
mobile +30 6945 107477
www.eikonika.net www.aim.cat
*************************************

Depending of course on the scale, this looks a lot like a bunch of
fungus spores, similar although not identical to:

http://www.mta.ca/dmf/download/jme/092310_0016.bmp

Maybe somebody was eating a bacon-mushroom burger in the microtomy area ;-)

Jim

--

James M. Ehrman
Digital Microscopy Facility
Mount Allison University
63B York St.
Sackville, NB E4L 1G7
CANADA

phone: 506-364-2519
fax: 506-364-2505
email: jehrman-at-mta.ca
www: http://www.mta.ca/dmf

A day without fusion is like a day without sunshine.

I agree, it does not looks like belonging to section. Some contamination
after sectioning/staining. I have stained a lot with PTA, have never seen
something like this.

Vladimir

Vladimir M. Dusevich, Ph.D.
Electron Microscope Lab Director
(816) 235-2072
371 School of Dentistry
650 E. 25th Street
Kansas City, MO 64108-2784

-----Original Message-----
X-from: nizets2-at-yahoo.com [mailto:nizets2-at-yahoo.com]
Sent: Thursday, January 23, 2014 8:23 AM
To: Dusevich, Vladimir

Hello Everyone,
Years ago, maybe 20, I saw SEM images of material observed in diesel exhaust. It's not carbon black, but we never did figure out what it was. It look very much like these images. Any chance it could be environmental contamination?

I seriously doubt its pollen. Most pollen to too, much too big to see in it's entirety in a TEM.

I look forward to finding out what it is!

Frank

-----Original Message-----
X-from: maryard-at-uga.edu [mailto:maryard-at-uga.edu]
Sent: Thursday, January 23, 2014 9:18 AM
To: Frank Karl

Panos,

What is the average size of the "particles"? You do not have a size bar in your image to get a feel for their size, which is important in any viral identification. The "particles" do not appear to be associated with the sample on the grid but laying on top of the sample. In my opinion, they look more like crystalline structures rather than viral particles. Depending on their actual size, could they possibly be phosphotungstic acid crystals?

Kind regards,
Mary

Mary Ard
Electron Microscopy Lab Coordinator
Department of Pathology
College of Veterinary Medicine
University of Georgia
501 D.W. Brooks Drive
Athens, GA 30602
706-542-5537

==============================Original Headers==============================
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Both spores and pollen are a way too big to be transparent under TEM beam.

Vladimir

Vladimir M. Dusevich, Ph.D.
Electron Microscope Lab Director
(816) 235-2072
371 School of Dentistry
650 E. 25th Street
Kansas City, MO 64108-2784

-----Original Message-----
X-from: jehrman-at-mta.ca [mailto:jehrman-at-mta.ca]
Sent: Thursday, January 23, 2014 9:22 AM
To: Dusevich, Vladimir

Depending of course on the scale, this looks a lot like a bunch of fungus spores, similar although not identical to:

http://www.mta.ca/dmf/download/jme/092310_0016.bmp

Maybe somebody was eating a bacon-mushroom burger in the microtomy area ;-)

Jim

--

James M. Ehrman
Digital Microscopy Facility
Mount Allison University
63B York St.
Sackville, NB E4L 1G7
CANADA

phone: 506-364-2519
fax: 506-364-2505
email: jehrman-at-mta.ca
www: http://www.mta.ca/dmf

A day without fusion is like a day without sunshine.

On 23/01/2014 11:05 AM, DusevichV-at-umkc.edu wrote:
}
}
} ----------------------------------------------------------------------
} ------ The Microscopy ListServer -- CoSponsor: The Microscopy Society
} of America To Subscribe/Unsubscribe --
} http://www.microscopy.com/MicroscopyListserver
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}
} I agree, it does not looks like belonging to section. Some contamination after sectioning/staining. I have stained a lot with PTA, have never seen something like this.
}
} Vladimir
}
} Vladimir M. Dusevich, Ph.D.
} Electron Microscope Lab Director
} (816) 235-2072
} 371 School of Dentistry
} 650 E. 25th Street
} Kansas City, MO 64108-2784
}
} -----Original Message-----
} X-from: nizets2-at-yahoo.com [mailto:nizets2-at-yahoo.com]
} Sent: Thursday, January 23, 2014 8:23 AM
} To: Dusevich, Vladimir
} Subject: [Microscopy] Fw: Virus
}
}
}
}
} ----------------------------------------------------------------------
} ------ The Microscopy ListServer -- CoSponsor: The Microscopy Society
} of America To Subscribe/Unsubscribe --
} http://www.microscopy.com/MicroscopyListserver
} On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} ----------------------------------------------------------------------
} ------
}
}
} Hi Panos!
}
} Giving a picture without scale to a miscroscopist is like talking without words. We cannot make anything out if it!
} I doubt these objects are embedded in the resin. One can see a hole in the section and one object seems to partly cover it. Also they don't look like they were sectioned.
} Looks like artifacts for me.
}
} Regards,
} Stephane
}
}
} ________________________________
} X-from: "pveril-at-apae.uth.gr" {pveril-at-apae.uth.gr}
} To: nizets2-at-yahoo.com
} Sent: Thursday, January 23, 2014 11:29 AM
} Subject: [Microscopy] Virus
}
}
}
}
}
} ----------------------------------------------------------------------
} ------ The Microscopy ListServer -- CoSponsor: The Microscopy Society
} of America To Subscribe/Unsubscribe --
} http://www.microscopy.com/MicroscopyListserver
} On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} ----------------------------------------------------------------------
} ------
}
}
} Dear all
}
} During an electron microscope examination of a Sparus aurata liver
} collagen (PTA and uranyl acetate were used as stains) I came across to
} these objects. They are probably virus particles. Can any one identify
} them?
}
} http://upload.users.uth.gr/files/virus.jpg
}
} Many thanks
} Panos

==============================Original Headers==============================
14, 21 -- From jehrman-at-mta.ca Thu Jan 23 09:21:14 2014 14, 21 -- Received: from smtpy.mta.ca (smtpy.mta.ca [138.73.1.143])
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Yes, please, we need a scale bar.

I take it that these are sitting on a film and are negatively stained?
With BOTH UrAc and PTA? They look vaguely like something tentatively
identified as scales from some sort of phytoplankton that we often see in
marine preps. They do not look like any of the many marine viruses we've
seen here, but I will show them to someone who works on marine viruses and
see what they say. However, right now I'm going with the idea that they
are stain crystals!

Aloha,
Tina

} During an electron microscope examination of a Sparus aurata liver
} collagen (PTA and uranyl acetate were used as stains) I came across to
} these objects. They are probably virus particles. Can any one identify
} them?
}
} http://upload.users.uth.gr/files/virus.jpg
}
} Many thanks
} Panos
} --
} Dr. Berillis Panagiotis
} Lecturer in Microscopy, image analysis and histology
} Department of Ichthyology and Aquatic Environment
} Agricultural school
} University of Thessaly
}
}
} ==============================Original Headers==============================
} 6, 18 -- From pveril-at-apae.uth.gr Thu Jan 23 04:23:12 2014
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****************************************************************************
* Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu *
* Biological Electron Microscope Facility * (808) 956-6251 *
* University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf*
****************************************************************************

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Yes, pollen was my immediate thought, but pollen would be at least one,
probably two, order/s of magnitude too big!

Dr Rosemary White
CSIRO Plant Industry
GPO Box 1600
Canberra, ACT 2601
Australia

T 61 2 6246 5475
F 61 2 6246 5334
E rosemary.white-at-csiro.au

On 24/01/14 4:02 AM, "eikonika-at-otenet.gr" {eikonika-at-otenet.gr} wrote:

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Thank you for the intense & helpful discussion!

We agree with those people who think that the "bubbles" have to do with
something extracted from the bacteria that prevents the resin from
polymerising.

We did not use osmium, we dehydrated in ethanol up to 80% ethanol and
did not degas the resin. We used a premix resin that works OK with any
other specimens except bacteria. But we do not add the extra accelerator.

Perhaps the next steps will be:
(i) to dehydrate up to 90 % alcohol
(ii) degas the resin
(iii) add extra accelerator so that whatever chemical reaction we have
might not occur?
Any suggestions are welcome and thank you again!

Stefan

==============================Original Headers==============================
5, 30 -- From stefan.wernitznig-at-medunigraz.at Fri Jan 24 04:43:33 2014
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Since bacteria are usually plentiful and inexpensive, I would also do one sample after 100% ethanol to see if it is do to residual water. Degassing the resin can change composition since you can differentially extract the more volatile components. Tom Phillips

Sent from my iPad

} On Jan 24, 2014, at 4:44 AM, "stefan.wernitznig-at-medunigraz.at" {stefan.wernitznig-at-medunigraz.at} wrote:
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} Thank you for the intense & helpful discussion!
}
} We agree with those people who think that the "bubbles" have to do with
} something extracted from the bacteria that prevents the resin from
} polymerising.
}
} We did not use osmium, we dehydrated in ethanol up to 80% ethanol and
} did not degas the resin. We used a premix resin that works OK with any
} other specimens except bacteria. But we do not add the extra accelerator.
}
} Perhaps the next steps will be:
} (i) to dehydrate up to 90 % alcohol
} (ii) degas the resin
} (iii) add extra accelerator so that whatever chemical reaction we have
} might not occur?
} Any suggestions are welcome and thank you again!
}
} Stefan
}
} ==============================Original Headers==============================
} 5, 30 -- From stefan.wernitznig-at-medunigraz.at Fri Jan 24 04:43:33 2014
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Email: tomw-at-uidaho.edu
Name: Tom Williams

Organization: Univ of Idaho

Title-Subject: [Filtered] TEM Diffraction of amphibole fibers and fiber bundles

Message: Question for any Mineralogist-Microscopist (or the other way 'round :) ) in the group.

I have a Mineralogy student working in my lab on the TEM trying to obtain diffraction patterns
(SAED) on amphibole fibers and fiber bundles (note: some of the specimens may be cleavage
fragments). Particle size: 50-100 microns short dimension & 100s if microns in the long D. Scope
JEOL 2010 at 200kV.

We are having a surprising difficult time getting any reasonable SAED patterns on these. We can get
reasonable and nice patterns on adjacent micas in the sample but nothing of use on the amphiboles.
Different spot sizes, SAED apertures, camera lengths, etc. No joy.

We should be able to do this-even though I am not the worlds best at electron diffraction?? I'm at
a loss and throwing myself Forrest Gump like asking for mercy!

Suggestions would be welcome.

Thanks-Tom

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Dear Tom,
50-100 µm is very thick for SAED. You are probably not getting the electrons through the sample, whereas, the micas are probably thinner. The thing to try is getting diffraction patterns from the very thinnest edges of the fibers. You could crush the sample further, but that would destroy the measurement of length-width aspect. If you have EDX, you can correlate composition of the crushed sample patterns to as-received samples.
Hope this helps.
Ken

|_|"|_|"|_|_|"|_|"|_|"|_|_|"|_|"|_|_|"|_|"|_|"|_|_|
Kenneth JT Livi, PhD
Director, The High-Resolution Analytical Electron Microbeam Facility
of the Integrated Imaging Center
Departments of Earth and Planetary Sciences and Biology
Olin Hall
3400 N Charles Street
Johns Hopkins University
Baltimore, Maryland 21218 USA
| | | .| | .| | .| | .| | | :| | | .| | .| | .| | .| | | :|

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Hi Tom,

the samples are thin enough for TEM? Your description sounds a lot like
the fibers are too thick, they need to be less than half a micron thick
to allow you a chance to get results. Cleavage is a great technique but
you might need to do it twenty times to get something decent.

Good luck
Rob

--
Robert Keyse, DPhil
Research Scientist
Lehigh University

5, East Packer Avenue,
Whitaker Laboratory
Bethlehem,
PA 18015-3194

Office (610)758-3465
Fax (610)758-4244

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Stefan,

I do not remember seeing how long your steps were during dehydration in
ethanol but do know that these little bacteria take longer than
anticipated to dehydrate. I have also found it advantageous to have the
ethanol moving by putting the tubes on a tilting table or to rotate them
during dehydration and also for the infiltration steps.

I have had similar looking bacteria in Macrophages embedded in EPON in the
past. After the first paper was published I learned that my problem was
their dehydration. The macrophages themselves looked fine when processed
by my standard tissue culture protocol but the Listeria within them turned
very dark and peppered like yours when the beam hit them. After I started
to use our longer protocol for tissue samples, the bacteria looked great.

Many years ago I was advised to go through 90% ethanol when using LR White
so your suggestion to do that is a good one.

One can introduce air/oxygen into the embedding media when stirring in the
accelerator so degassing could help in the elimination of that possibility.

I usually harden the LR White in a 45 degree C. oven for a few days
without accelerator so I can not address the possibility of adding an
additional amount of accelerator.

Opinions and experiences related are those of Pat Connelly and do not
represent the NIH.
This message is not confidential and can be freely shared and reproduced.

Pat

Patricia Connelly
Research Assistant
NHLBI Electron Microscopy Core
National Institutes of Health
14 Service Road West
Bldg. 14E Rm. 111B MSC 5570
Bethesda, MD 20892-5570
Phone 301-496-3491
FAX 301-402-0170
connellyps-at-nhlbi.nih.gov

On 1/24/14 5:56 AM, "stefan.wernitznig-at-medunigraz.at"
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Tom,

In bright field look for areas that exhibit bend contours. Those are the areas that are thin enough where you can find a reasonable pattern usually at the terminations.

Find fibers that are elongated in the direction of the tilt axis, adjust the height of the sample so you can tilt without the fiber moving.

Tilting about the long axis which is [001] you can see the closely spaced spots of 010.

Tilting about 010, ideally you should find [100] and [101] about 30° apart.

Easy to find in the orthorhombic amphiboles and less easy in the monoclinic amphiboles.

I suggest practicing with iucc standards. Tremolite and anthophyllite.

Also with Dave Palmer's Single Crystal program you don't need a TEM.

Getting indexable diffraction patterns off fibrous amphiboles requires patience which is why asbestos labs don't do it.

Gordon
---------------------------------------------------
Gordon L. Nord, Ph. D
Senior Scientist
International Environmental Research Foundation
P.O. Box 3459, Grand Central Station
New York, NY 10163-3459

{http://www.ierfinc.org/}

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Dear Panos,

The objects you have seen under your EM are not virus or pollen, They are Brochosomes.
Brochosomes are secretory granules produced by the Malphigian tubules of leafhoppers.
You may have a look at the Wikipedia article concerning this unfamiliar but beautiful structure (wikipedia.org/wiki/Brochosome) .
I have seen EM pictures of this structure many years ago , since it has been once described in the sixties by my former colleagues in the lab in Rennes (France, Brittany).
(Gouranton J. & Maillet P.L. (1967) Origine et structure des brochosomes. Journal de Microscopie 6: 53-64.).
Brochosomes are source of contaminations and can sometimes be found in aerosol
(Wiffen R. D. & Heard M. J. (1969) Unidentified airborn species. Nature 224: 715).
I recommend the readings associated with the Wiki article. However, if you are interested, I can look over the archives of the lab and try to find original negatives.

Daniel

Daniel THOMAS
UMR CNRS 6290
Universite de Rennes 1
35042 RENNES, FRANCE

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Title-Subject: [Filtered] RJ LEE PSEM MANUAL WANTED

Message: Hi all. I would be very grateful to anyone helping me to find a copy (either original,
photocopy or .pdf) of the RJ LEE PERSONAL SEM - PSEM, earlier model.
Users impressions and tips also welcome.
Looking forwartd receiving news from you, I remain.
Best regards
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Merci Daniel for showing these brochosomes, they have a very smart shape. A
pattern repeated in many structures seen at different scales in this
wonderful world. And I am sure you gave the right answer.

yorgos

Dr Yorgos Nikas
Athens Innovative Microscopy
Skra 36 Voula 16673 GREECE

Tel/fax +30 210 8957677
mobile +30 6945 107477
www.eikonika.net www.aim.cat
*************************************

----- Original Message -----
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Sent: Saturday, January 25, 2014 1:55 PM

March 24-28, 2014 a course in scanning electron microscopy will be offered at Hooke College of Applied Sciences.
Further information about this hands-on course can be found at the link below:

http://www.hookecollege.com/courses/course.asp?COURSE_ID=42

Best regards-
__________________________________________________
Chris Gorman
Admissions Specialist
Hooke College of Applied Sciences
850 Pasquinelli Drive
Westmont, IL 60559-5539 USA
630-887-7100 (tel)
630-887-7412(fax)
www.hookecollege.com

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Test I told you not to open...........

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From medications_canadian7-at-cyfrowypolsat.pl Tue Jan 28 11:58:16 2014
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We have an old Zygo optical profilometer that we are looking to get rid of.
The microscope works fine but the computer to which it is attached is not
operational as no one remembers the root password and we don't have the
operating system tapes to re-install from, it runs HP Unix. I have taken
some photos and they can be viewed at;

https://netfiles.umn.edu/xythoswfs/webui/_xy-e16168960_1-t_UXIaLHg1

Again the item is in Minneapolis, MN and it is free but the buyer must
assume all shipping costs.

Nick Seaton
University of Minnesota
Characterization Facility

--

Dr Nicholas Seaton

SEM Specialist
Characterization Facility

College of Science and Engineering

University of Minnesota

Shepherd Labs

100 Union Street SE

Minneapolis

MN, 55455

email: seato008-at-umn.edu

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From pharmacy_reasonable7-at-kidsforsavingearth.org Wed Jan 29 01:07:16 2014
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Name: Ravi

Title-Subject: [Filtered] Role of CaCl2 in biological sample fixation for TEM

Message: What is the role of CaCl2 in biological sample fixation for TEM.?
What is the recommended concentration required in phosphate buffer.?

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Ravi,
Calcium and magnesium chloride are used in TEM fixatives to protect membranes from enzymes that are still active during fixation. This is refered to as maintaining membrane tonicity (small breaks in membranes may occur). If phosphate buffers are used, you must use only MgCl2 (3 mM), as calcium will precipitate to calcium phosphate.

sincerely,
Michael Delannoy
Johns Hopkins SOM Microscope Facility

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Title-Subject: [Filtered] Diamond knife sharpening

Message: Has anyone out there actually seen how the diamond knives are resharpened? I wonder if
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us. Anyone also know of a compnmay that takes several knives in exchange for a knife? Like giving up
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Group

Working on an issue that has come up in my Leo 1450VPSE SEM since I changed a gun filament.
Seeing light and dark scan lines in captured images. First thought it's some kind of charging behavior but not sure.
Problem occurs with both SE and BS detectors as I try to capture an image using the LINE AVG mode where a line of the image is scanned a number of times (N) before moving to the next line. This builds my image one scan line at a time till I have a full frame.
Using scan speed six (rate it scans frame) I get different widths of shadow line as I change N.

Since I cannot attach images, I can Email them direct to anyone that may want to offer a suggestion as to what is causing the problem.

Thanks

Roy Beavers
Southern Methodist University
rbeavers-at-smu.edu

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Nick,
Diatome diamond knives will replace a used knife (any size) for a brand new knife (any size) at half the cost of a new knife. Believe me, this is the best deal I know of. If you find a better one, let us all know.

sincerely,
Michael Delannoy
Johns Hopkins SOM Microscope Facility

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Nick,

Years ago, a knife vendor gave a talk on how diamond knives are sharpened. It is not hard, but takes 1-2 weeks of grinding on a diamond grinding wheel. This in not something that you can do in your leisure. I once asked another vendor this same question about giving them 3 or four knives and get a new sharpened knife in return. After picking themselves off the ground from laughing so hard, they explained to me that the diamond is about 40-50 dollars, it is the time it takes to sharpen the knife that cost so much, so they will not just give a sharpened knife with exchange of 3 or 4 knives. One of the vendors does give a discount for a 2 for 1 exchange. Call them up and ask about this.

Best of luck convincing the powers that be to purchase a knife for you.

Ed

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Title-Subject: [Filtered] Re: viaWWW:Diamond knife sharpening

Message: Diatome has a deal where if you send in three knives for re-sharpening, they will charge
for two of them at the re-sharpen price and the third one is free.

Since you have many knives it might be advantageous to keep a few sets of three so when you send one
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Thank you for you detailed answers!

From most answers - especially Pat's answer - I gather that dehydration
is important, so our next test will be whether dehydrating to 90 or 100
% ethanol improves the structure and gets rid of the "bubbles". The
alcohol steps were for 30 min each with an overnight break in 70 %
alcohol so I do not think longer incubation times will help.

We use LR white because we want to perform immunolabelling, so I think
we have to find the right balance between structural preservation and
preservation of antigenicity. Our next approach will be with dehydration
up to 80, 90, and 100 % ethanol in parallel.
Thank you again!

Stefan

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O.k., higher ups at the University have been looking at service
contracts costs and trying to

Richard E. Edelmann, Ph.D., Director
Center for Advanced Microscopy & Imaging
9C Upham Hall
Miami University, Oxford, OH 45056
Ph: 513.529.5712 Fax: 513.529.4243
E-mail: edelmare-at-miamioh.edu
http://www.cami.muohio.edu

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(Apologies for slippery fingers)

O.k., higher ups at the University have been looking at service
contracts costs and trying to find cost reductions.
One the vendors they are now thinking about is Unity Asset Management
Service (Part of Thermo Fisher Scientific). To provide repair services for all
the equipment on campus.

I did a search of the listserv archives and did not find any thing about Unity.
SO does anyone out there have any feed back with regards to "Unity Asset
Management Service"? For Microscopes or anything else.

I do not want to start a war, and I know alternatoves have been disscussed
before, but its new year and time to discuss a little again.

Happy to take anything offline or through the list or even a phone call.

Thank you.

Richard E. Edelmann, Ph.D., Director
Center for Advanced Microscopy & Imaging
9C Upham Hall
Miami University, Oxford, OH 45056
Ph: 513.529.5712 Fax: 513.529.4243
E-mail: edelmare-at-miamioh.edu
http://www.cami.muohio.edu

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To All:

I hope all is well. If you are interested in advancing your EELS and EFTEM
(analytical TEM) knowledge, Gatan is offering a 4 day professional
training school on April 8-11, 2014 at its R&D Headquarters in Pleasanton,
CA, USA. Here are all the details and online registration:
http://www.gatan.com/resources/training/EELSSchoolApr2014.php

--
Best regards,

John
Marketing Communications Manager

Gatan, Inc.

Email: info-at-gatan.com
Website: www.gatan.com

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dear Rick, dear all,

} O.k., higher ups at the University have been looking at service
} contracts costs and trying to find cost reductions.

every year the same issue, in your institution and in many/most others as well.

from my 23 years experience, a service contract over 23 years is far better for the scopes and for all users of the scopes (i.e. for all projects!) than 23 years of waiting for the engineer(s), not knowing when (s)he will come, and who is going to pay for it. - We all know this, and most people in the community will agree - except some / many of the "Ups at the University" (nice word...), and those in the Administration.

} One the vendors they are now thinking about is Unity Asset Management
} Service (Part of Thermo Fisher Scientific). To provide repair services for
} all the equipment on campus.

for most of us: the service engineers of the companies are best trained on the scopes of each particular brand. Thus, a service contract for an electron microscope, and for most of the modern complex CLSM's and their variants, is "best-value-for-money", no doubt about this - in most cases. I am aware that there are other meanings around, other experiences; like: no good experience with the engineer, or too-few-engineers ... (happened to me as well, several times), or trained engineers are available / employed in the institution / University: Not here.

} I did a search of the listserv archives and did not find any thing about Unity.
} SO does anyone out there have any feed back with regards to "Unity Asset
} Management Service"? For Microscopes or anything else.
I do not know anything about Unity, sorry.

One of my arguments. Let's take an (ordinary) car for 25.000 EUR or $ (not really comparible to our scopes, in complexity and pricing!) which is used daily, say 6 hours per day. Quickly, you will end up with an annual mileage of 20.000 km at least, if not 50.000 km (ask your service engineer, who really drive a lot!). This will raise maintenance costs for this car (oil, tyres, repairs, ...) of at least 1.000 EUR or $ per year (i.e. 4% of the initial investment), most likely even higher. Not in the first two years, but then, it will increase, quickly, dramatically. We are prepared to pay for this, because a car is what we think we need, daily. - Our students / postdocs need the scopes daily, probably more than the car (most likely), and in many institutions, the scope is far more complex and used for more hours than only 6 per day. - But, 3 to 4% of the investment for the annual maintenance contract, offering quick and expert response: this is too high, too much money? - And, compare this with the vendor of your car ... are the service engineers as good as those for the scopes? do they travel such long distances, each week / month?
Richard, I am facing the same questions, every year or every second year.
I still hope that we can convince the people "Up-in-the-Univ" or in the Administration.

kind regards,
Reinhard

--
Prof. Dr. Reinhard Rachel
University of Regensburg
Centre for EM / Anatomy
Faculty of Biology & Preclin. Med.
Universitaetsstrasse 31
D-93053 Regensburg - Germany
tel +49 941 943 2837, 1720
fax +49 941 943 2868
mail reinhard.rachel-at-biologie.uni-regensburg.de
office: VKL 3.1.29

Next microscopy conference:
http://www.imc2014.com/
18th IMC 2014 in Prague, Czech Rep.

==============================Original Headers==============================
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Email: fxwatanabe-at-ualr.edu
Name: Fumiya Watanabe

Organization: Center for Integrative Nanotechnology Sciences UALR

Title-Subject: [Filtered] Follow up on Consolidated Service Contract inquiry

Message: Hello everyone who are concerned with similar problem of
pressure to consider a consolidated service contract.

There was a post by Richard Edelmann on Unity Asset Management Service.
We are being urged to consider another service called Remi (state of
Arkansas has a contract with them, I've been told):

https://www.theremigroup.com/default.aspx

Our main instruments are from JEOL (TEM/SEM), Bruker (XRD and AFM),
Horiba (Raman, Photoluminescence, Elipsometry), Thermo Fisher (XPS),
etc. Does anybody have experience with Remi service (especially with
cases involving other vendors such as JEOL, Bruker-AXS, or Thermo
Fisher)? I have contacted Richard and have received reply from him
already on Unity (thank you very much Richard for very quick reply). Any
information on Remi would help us since we have no idea about them.

Thank you very much.

Fumiya

Fumiya Watanabe, Ph.D.
Director of Instrumentation
Center for Integrative Nanotechnology Sciences
ETAS 153E
University of Arkansas at Little Rock
2801 S. University Ave.
Little Rock, AR 72204
(501) 683-7479 (Office)

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Name: Pete Finger

Organization: The Jackson Laboratory

Title-Subject: [Filtered] Negative Staining Problem

Message: IÂ’ve been doing some negative staining and am experiencing some
problems with the Formvar film falling apart on the grid. IÂ’ve been
using 200 mesh Copper Formvar/carbon coated grids which are a couple of
years old Â– does anyone know if these grids have a certain shelf life?
My technique has been to put a drop of the protein (in HEPES buffer)
on the grid, blot dry, add a drop of uranyl acetate, blot dry and then
let the grid air dry. When I view the grid on the TEM, pretty much all
the Formvar is gone. I havenÂ’t done a lot of negative staining in the
past so any comments/suggestions would be greatly appreciated. Thanks much.

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From canadian_drugs15-at-mgts.by Fri Jan 31 14:43:34 2014
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Dear all,
dear Nestor,
dear Fumiya,

in 2012 there was a thread on MSA-Listserver concerning the matter you are requesting:

Thread started 08.05.2012 - ended: 09.06.2012
"Remi Group as a equipmt service contract provider"

There have been several answers.
For your convenience (so you would not need to got through the MSA-Listserver Archives):

Since I was interested too, I have collected these Replies in a word.docx/pdf, which I could provide easily in sending to your e-mail-box, if you allow this.
Just write to me at w.muss-at-salk.at and receive - with your permission the pdf of this collection of replies.

Best wishes and regards,

Wolfgang

Wolfgang MUSS PhD
EM-Lab
Univ. Inst. Pathology,
SALK-LKH(Gen. Hospital) & PMU (Private Paracelsus Medical University) Salzburg
SALZBURG - AUSTRIA

Von: zaluzec-at-microscopy.com [mailto:zaluzec-at-microscopy.com]
Gesendet: Freitag, 31. Jänner 2014 20:11
An: Muß Wolfgang
Betreff: [Microscopy] Re(Fwd): Unity Asset Management Service? [by companies, service contracts costs and trying to find cost reductions] // REMI-Service: Follow up on Consolidated Service Contract inquiry

[Microscopy] !! Re: Unity Asset Management Service? [by companies, service contracts costs and trying to find cost reductions] !!
// Follow up on Consolidated Service Contract inquiry

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Hello everyone who are concerned with similar problem of pressure to consider a consolidated service contract.

There was a post by Richard Edelmann on Unity Asset Management Service.
We are being urged to consider another service called Remi (state of Arkansas has a contract with them, I've been told):

https://www.theremigroup.com/default.aspx

Our main instruments are from JEOL (TEM/SEM), Bruker (XRD and AFM), Horiba (Raman, Photoluminescence, Elipsometry), Thermo Fisher (XPS), etc.
Does anybody have experience with Remi service (especially with cases involving other vendors such as JEOL, Bruker-AXS, or Thermo Fisher)?

I have contacted Richard and have received reply from him already on Unity (thank you very much Richard for very quick reply).
Any information on Remi would help us since we have no idea about them.

Thank you very much.

Fumiya

Fumiya Watanabe, Ph.D.
Director of Instrumentation
Center for Integrative Nanotechnology Sciences
ETAS 153E
University of Arkansas at Little Rock
2801 S. University Ave.
Little Rock, AR 72204
(501) 683-7479 (Office)

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From boakkeau-at-wegl.net Mon Feb 3 09:59:44 2014
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Message-Id: {201402031559.s13FxfSr020679-at-ns.microscopy.com}

Pete,

Films on grids do tend to break when old. I used some 75 mesh grids that
were 5 years old recently and they were still good but I'd not try to use
single holed ones that were that old unless I really needed to. With
older 200 mesh grids one can usually get some areas that do not pop but
use newer ones if you can afford them or make them yourself. Perhaps your
drying step is a bit harsh. The word "blot" is what caught my eye. It is
hard to judge unless it could be observed.

I was taught to do my negative staining differently than how you describe.
I put a sample onto a recently glow discharged grid held in forcepts, let
it sit for 30 to 60 seconds, gently drop 5-6 drops of 1%UA from a pipet
onto the surface of the grid held perpendicular to the grid (grid parallel
to the table top over a vessel to catch the UA). After the last drop of UA
is on the grid I touch a point of filter paper to the junction of the
forcepts where the grid is being held to slowly wick the UA off. The
surface of the grid will still be wet. There is usually a small amount of
UA between the forcept tips so I "push" the grid out of the forcepts with
the same piece of filter paper using another portion that is not already
wet, onto some lens tissue to continue to dry completely.

There are many different techniques in doing negative staining. Once you
find one that works for you that is the one you should stick with.

Opinions and experiences related are those of Pat Connelly and do not
represent the NIH. This message is not confidential and can be freely
shared and reproduced.

Pat

Patricia Stranen Connelly
NHLBI EM Core Facility
National Institutes of Health
Building 14E Room 111B
Bethesda, MD 20892-5570
301-496-3491

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Email: pete.finger-at-jax.org
Name: Pete Finger

Organization: The Jackson Laboratory

Title-Subject: [Filtered] Negative Staining Problem

Message: I?ve been doing some negative staining and am experiencing some
problems with the Formvar film falling apart on the grid. I?ve been
using 200 mesh Copper Formvar/carbon coated grids which are a couple of
years old ? does anyone know if these grids have a certain shelf life?
My technique has been to put a drop of the protein (in HEPES buffer)
on the grid, blot dry, add a drop of uranyl acetate, blot dry and then
let the grid air dry. When I view the grid on the TEM, pretty much all
the Formvar is gone. I haven?t done a lot of negative staining in the
past so any comments/suggestions would be greatly appreciated. Thanks much.

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Title-Subject: [Filtered] 200KV used TEM wanted

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We are looking for am used TEM, 200-300KV not FEG gun.
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Email: 12hy1-at-queensu.ca
Name: Hongbing Yu

Organization: Queens' University, Canada

Title-Subject: [Filtered] EDS can't detect tin

Message: We are trying to use FEI TECNAI ORISIS TEM characterize the
element distribution of a Zirconium alloy with 3.5 atom% Sn. However we
can't detect tin anywhere with EDS in STEM Mode with beam spot 9. The
EDS system works very well for a ODS steel. We have tried our best to
figure out this problem, but we failed. Any comment and suggestion would
be appreciated.

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Dear Hongbing,

This may sound like a silly suggestion but if you haven't already, I
would try the following things:

1) Use an enormous beam in TEM mode to look around on micron-scale and
look for Sn in EDS all over any thin area. You may just be looking at
an unusual part of the specimen where the Sn is gone and not know it.

2) If you can do EELS, check for Sn there - should be an M edge about
500eV I think (look in EELS Atlas, I am being vague). If you cannot
see Sn anywhere by EELS or EDS then the problem is not the EDS
detector, the problem is that your specimen doesn't have Sn in it, and
you now have to figure out why. (Preferential removal of Sn during
sample prep? Something happened when making the material originally?)

Good luck!

Yours

Jo Sharp

On 4 February 2014 21:16, {zaluzec-at-microscopy.com} wrote:
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} Title-Subject: [Filtered] EDS can't detect tin
}
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} element distribution of a Zirconium alloy with 3.5 atom% Sn. However we
} can't detect tin anywhere with EDS in STEM Mode with beam spot 9. The
} EDS system works very well for a ODS steel. We have tried our best to
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} 14, 39 -- Subject: viaWWW:EDS can't detect tin
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March 3-7, 2014 a course in polarized light microscopy will be offered at Hooke College of Applied Sciences.
Further information about this hands-on course can be found at the link below:

http://www.hookecollege.com/courses/course.asp?COURSE_ID=41

Best regards-
__________________________________________________
Chris Gorman
Hooke College of Applied Sciences
850 Pasquinelli Drive
Westmont, IL 60559-5539 USA
630-887-7100 (tel)
630-887-7412(fax)
www.hookecollege.com

==============================Original Headers==============================
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Email: nxa157-at-case.edu
Name: Nanthawan Avishai

Organization: MSNO

Title-Subject: [Filtered] MSNO winter meeting -Feb 26,2014

Message: MSNO Winter MEETING will be on Wednesday, February 26th, 2014,
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Dear All,

anybody out there to help me with a service manual and a parts list for
the Reichert Ultracut E ultramicrotome?
All I have is the user manual.

Thanks,
Stefan

--

-----------------------------------------------------
Stefan Diller - Scientific Photography
Arndtstrasse 22
D - 97072 Wuerzburg Germany
++49-931-7848700 Phone
++49-931-7848701 Fax
++49-175-7177051 Mobile

Websites:
www.nanoflight.info
www.electronmicroscopy.info
www.elektronenmikroskopie.info
www.assisi.de
www.zwillingsprojekt.de
www.stefan-diller.com
Anfahrt: http://Mail.map24.com/Stefan.Diller
-----------------------------------------------------

==============================Original Headers==============================
9, 20 -- From stefan.diller-at-t-online.de Thu Feb 6 03:57:17 2014
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Hi Listers

I was just sent some material on a new microscope from Seron and it
occurred to me I have not really heard much on the list about this
brand. Has anyone tried one or have one even? How do they fair to
other entry level micorscopes. Comments on or off list would be
appricated.

Thanks
John in the UK (Bath Uni)

==============================Original Headers==============================
3, 26 -- From john.mitchels-at-gmail.com Thu Feb 6 07:47:47 2014
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3, 26 -- Subject: Seron SEMs any views?
3, 26 -- From: John Mitchels {john.mitchels-at-gmail.com}
3, 26 -- To: Microscopy-at-microscopy.com
3, 26 -- Content-Type: text/plain; charset=ISO-8859-1
==============================End of - Headers==============================

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Dear All,
thanks, I finally got a PDF of the service manual.

Stefan

-----------------------------------------------------
Stefan Diller - Scientific Photography
Arndtstrasse 22
D - 97072 Wuerzburg Germany
++49-931-7848700 Phone
++49-931-7848701 Fax
++49-175-7177051 Mobile

Websites:
www.nanoflight.info
www.stefan-diller.com
www.electronmicroscopy.info
www.elektronenmikroskopie.info
www.zwillingsprojekt.de
www.assisi.de
Anfahrt: http://Mail.map24.com/Stefan.Diller
-----------------------------------------------------

Am 06.02.14 14:28, schrieb Muß Wolfgang:
} bradford.ross-at-botany.ubc.ca {mailto:bradford.ross-at-botany.ubc.ca}
}
} Stefan:
}
} ich bin ja schon sehr froh, wenn ich als „Notnagel“ wenigstens da und dort einmal was „tun“ kann für die Community....Für Dich
} allemal von Herzen!
}
} Habe noch was gefunden (originales pdf hatte 13 MB, daher habe ich es nochmals als pdf „gedruckt“ und damit auf ca. 7MB
} komprimiert...das hat etwas gedauert...)
}
} Die Dokumentation hatte ich dankenswerterweise von bradford.ross-at-botany.ubc.ca {mailto:bradford.ross-at-botany.ubc.ca} im Nov. 2013
} erhalten.
}
} Ebenfalls „sonnige“, und v. a. {föhnige} Grüße aus Salzburg (dzt. wie im späten April: knallblauer Himmel bei ca. 15Grad C!
}
} Wolfgang
}
} *Von:*Stefan Diller [mailto:stefan.diller-at-t-online.de]
} *Gesendet:* Donnerstag, 06. Februar 2014 14:21
} *An:* Muß Wolfgang
} *Betreff:* Re: persönl. AW(2): [Microscopy] Reichert Ultracut E service manual
}
} Lieber Wolfgang,
} wie immer der Retter in der Not ;-)
}
} Herzliche Grüße in den Süden,
} Stefan
}
} Am 06.02.2014 14:18, schrieb Muß Wolfgang:
}
} Lieber Stefan,
}
} das ist alles, was ich in meinen files gefunden habe....
}
} mhG
}
} und beste Wünsche beim Basteln...
}
} Wolfgang
}
} *Von:*stefan.diller-at-t-online.de {mailto:stefan.diller-at-t-online.de} [mailto:stefan.diller-at-t-online.de]
} *Gesendet:* Donnerstag, 06. Februar 2014 11:02
} *An:* Muß Wolfgang
} *Betreff:* [Microscopy] Reichert Ultracut E service manual
}
} ---------------------------------------------------------------------------
}
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} ----------------------------------------------------------------------------
}
} Dear All,
}
} anybody out there to help me with a service manual and a parts list for
}
} the Reichert Ultracut E ultramicrotome?
}
} All I have is the user manual.
}
} Thanks,
}
} Stefan
}
} --
}
} -----------------------------------------------------
}
} Stefan Diller - Scientific Photography
}
} Arndtstrasse 22
}
} D - 97072 Wuerzburg Germany
}
} ++49-931-7848700 Phone
}
} ++49-931-7848701 Fax
}
} ++49-175-7177051 Mobile
}
} Websites:
}
} www.nanoflight.info {http://www.nanoflight.info}
}
} www.electronmicroscopy.info {http://www.electronmicroscopy.info}
}
} www.elektronenmikroskopie.info {http://www.elektronenmikroskopie.info}
}
} www.assisi.de {http://www.assisi.de}
}
} www.zwillingsprojekt.de {http://www.zwillingsprojekt.de}
}
} www.stefan-diller.com {http://www.stefan-diller.com}
}
} Anfahrt: http://Mail.map24.com/Stefan.Diller
}
} -----------------------------------------------------
}
} ==============================Original Headers==============================
}
} 9, 20 -- From stefan.diller-at-t-online.de {mailto:stefan.diller-at-t-online.de} Thu Feb 6 03:57:17 2014
}
} 9, 20 -- Received: from mailout05.t-online.de (mailout05.t-online.de [194.25.134.82])
}
} 9, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id s169vG4m007656
}
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}
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}
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==============================Original Headers==============================
9, 23 -- From stefan.diller-at-t-online.de Thu Feb 6 08:43:56 2014
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I originally posted this as "free SEM" but Nestor's spam filter threw me
under the bus...

A friend of mine passed this ad on to me. I have no connection to the
people giving this away, just thought someone out there might be
interested. I'm not familiar with the make/model. Anybody? It does
appear to be "vintage"...

http://ottawa.kijiji.ca/c-buy-and-sell-tools-other-Scanning-Electron-Microscope-W0QQAdIdZ564766335

Cheers,

Jim

--

James M. Ehrman
Digital Microscopy Facility
Mount Allison University
63B York St.
Sackville, NB E4L 1G7
CANADA

phone: 506-364-2519
fax: 506-364-2505
email: jehrman-at-mta.ca
www: http://www.mta.ca/dmf

If you give some managers an inch they think they’re a ruler.

==============================Original Headers==============================
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Hello Microscopists,
Here at the University of Texas at El Paso we are searching for a new EM to purchase. We mainly use Hitachi SEM and TEM for Hard materials characterization. We are mainly searching for an EM with specialization in polymers characterization. In our search we stumbled upon the LEVM5 Benchtop TEM from Delong America for polymers. The unit claims to have TEM, SEM, and STEM capabilities. We here in the southwest were wondering what are/were your experiences the LVEM5 or other Delong EM’s? We were also wondering how are the products from the Delong America Company in the perspective of materials characterization? If anybody wouldn't mind sharing those experiences with us that would be great.

Thanks for your thoughts and answers in advance
Alejandro Hinojos

==============================Original Headers==============================
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6, 29 -- From: "Hinojos, Alejandro" {ahinojos2-at-miners.utep.edu}
6, 29 -- To: "Microscopy-at-Microscopy.Com" {Microscopy-at-Microscopy.Com}
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Hi Alejandro,

It is a 5kV instrument. For TEM (or STEM), ask yourself what thickness
you can look through with a 5kV beam.

Cheers,
Henk

On 2/6/2014 9:19 PM, ahinojos2-at-miners.utep.edu wrote:
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} Hello Microscopists,
} Here at the University of Texas at El Paso we are searching for a new EM to purchase. We mainly use Hitachi SEM and TEM for Hard materials characterization. We are mainly searching for an EM with specialization in polymers characterization. In our search we stumbled upon the LEVM5 Benchtop TEM from Delong America for polymers. The unit claims to have TEM, SEM, and STEM capabilities. We here in the southwest were wondering what are/were your experiences the LVEM5 or other Delong EM’s? We were also wondering how are the products from the Delong America Company in the perspective of materials characterization? If anybody wouldn't mind sharing those experiences with us that would be great.
}
}
} Thanks for your thoughts and answers in advance
} Alejandro Hinojos
}
}
}
}
} ==============================Original Headers==============================
} 6, 29 -- From ahinojos2-at-miners.utep.edu Thu Feb 6 20:17:12 2014
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Hi

A collegue was interested by this instrument last year and made some
tests. His goal was to have an easy access to a TEM like instrument, to
make fast controls in nanoparticules syntesis (organics with heavy
metals), as the acess to our TEMs need some days of delay.

Hearing about the price of that instrument, I proposed him to test too a
STEM device to be fitted on our FE-SEM.
Comparing prices and performences (principally the versatility), the
choice was easy. The STEM device for the SEM gives better images, the
possibility to choose the HV up to 30 keV and a price 8x less expensive...

The real adavantage of this table-top TEM I could see is that it can be
used easy by non-microcopists, like table-top SEMs, with on the other
hand the limitations of the undestanding such people will have of what
they do.

So if you soon have a (FE)SEM, it could be usefull to consider if the
STEM device for SEM would not be a better solution. But I'm interested
on what your tests will bring you as conclusions.

Hope it helps

J. Faerber
IPCMS-DSI
Institut de Physique et Chimie des Matériaux de Strasbourg
Département Surfaces et Interfaces
23, rue de Loess ; BP43
67034 Strasbourg CEDEX 2
France

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

Le 07/02/2014 03:28, ahinojos2-at-miners.utep.edu a écrit :
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}
} Hello Microscopists,
} Here at the University of Texas at El Paso we are searching for a new EM to purchase. We mainly use Hitachi SEM and TEM for Hard materials characterization. We are mainly searching for an EM with specialization in polymers characterization. In our search we stumbled upon the LEVM5 Benchtop TEM from Delong America for polymers. The unit claims to have TEM, SEM, and STEM capabilities. We here in the southwest were wondering what are/were your experiences the LVEM5 or other Delong EM’s? We were also wondering how are the products from the Delong America Company in the perspective of materials characterization? If anybody wouldn't mind sharing those experiences with us that would be great.
}
}
} Thanks for your thoughts and answers in advance
} Alejandro Hinojos
}
}
}
}
} ==============================Original Headers==============================
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10, 25 -- From jacques.faerber-at-ipcms.u-strasbg.fr Fri Feb 7 02:01:34 2014
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Hi

I have worked with a number of clients over the years, when we have used the
SEM in a STEM mode. The advent of FEG systems dramatically changed the
capabilities of the SEM to produce good quality STEM images, and the use of
these instruments in x-ray analysis has been very fruitful. We have
produced a paper that will be in the March "Microscopy Today", this
summarises some of the work we have been doing. The use of thin films for
x-ray analysis in the SEM, even without a STEM facility, is often a step
forward that would have been more difficult to make if a dedicated TEM were
used. There are many advantages to using a SEM for thin film x-ray
analysis. The spectrum is less likely to be "contaminated" by signals from
the instrument itself, and elemental mapping at high resolution is easily
accomplished.

Regards

Steve

Steve Chapman FRMS
Protrain for Consultancy and Courses in Electron Microscopy
Mobile +44 (0) 7711 606967 Fax +44 (0)1280 814007
www.emcourses.com

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realname - Alejandro Hinojos
Email - ahinojos2-at-miners.utep.edu
ORGANIZATION - University of Texas at El Paso
EDUCATION - Undergraduate College
LOCATION - El Paso
SUBJECT_OF_QUESTION - TEM Shopping advice
QUESTION - Hello Microscopists,
Here at the University of Texas at El Paso we are searching for a new EM
to purchase. We mainly use Hitachi SEM and TEM for Hard materials
characterization. We are mainly searching for an EM with specialization
in polymers characterization. In our search we stumbled upon the LEVM5
Benchtop TEM from Delong America for polymers. The unit claims to have
TEM, SEM, and STEM capabilities. We here in the southwest were wondering
what are/were your experiences the LVEM5 or other Delong EM's? We were
also wondering how are the products from the Delong America Company in
the perspective of materials characterization? If anybody wouldn't mind
sharing those experiences with us that would be great.

Thanks for your thoughts and answers in advance
Alejandro Hinojos

==============================Original Headers==============================
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Email: ahinojos2-at-miners.utep.edu
Name: Alejandro Hinojos

Organization: University of Texas at El Paso, Metalurgical & Materials Engineering Department

Title-Subject: [Filtered] EM Shopping

Message: Hello Microscopists,
Here at the University of Texas at El Paso we are searching for a new EM to purchase. We mainly use
Hitachi SEM and TEM for Hard materials characterization. We are mainly searching for an EM with
specialization in polymers characterization. In our search we stumbled upon the LEVM5 Benchtop TEM
from Delong America for polymers. The unit claims to have TEM, SEM, and STEM capabilities. We here
in the southwest were wondering what are/were your experiences the LVEM5 or other Delong EM’s? We
were also wondering how are the products from the Delong America Company in the perspective of
materials characterization. If anybody wouldn't mind sharing those experiences with us that would be
great.

Thanks for your thoughts and answers in advance
Alejandro Hinojos

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While I have a STEM/TEM, from time to time I use SEM/EDS for TEM grids with all advantages Steve mentioned. Just place several TEM grids with thin films on a carbon planchet, and go on! Remember: EDS resolution is determined mostly by thickness of specimen, not by instrument itself. So, EDS spatial resolution in SEM could be measured in nanometers (instead of microns) if thin specimens are used.

Vladimir

Vladimir M. Dusevich, Ph.D.
Electron Microscope Lab Director
(816) 235-2072
371 School of Dentistry
650 E. 25th Street
Kansas City, MO 64108-2784

-----Original Message-----
X-from: protrain-at-emcourses.com [mailto:protrain-at-emcourses.com]
Sent: Friday, February 07, 2014 4:57 AM
To: Dusevich, Vladimir

Hi

I have worked with a number of clients over the years, when we have used the SEM in a STEM mode. The advent of FEG systems dramatically changed the capabilities of the SEM to produce good quality STEM images, and the use of these instruments in x-ray analysis has been very fruitful. We have produced a paper that will be in the March "Microscopy Today", this summarises some of the work we have been doing. The use of thin films for x-ray analysis in the SEM, even without a STEM facility, is often a step forward that would have been more difficult to make if a dedicated TEM were used. There are many advantages to using a SEM for thin film x-ray analysis. The spectrum is less likely to be "contaminated" by signals from the instrument itself, and elemental mapping at high resolution is easily accomplished.

Regards

Steve

Steve Chapman FRMS
Protrain for Consultancy and Courses in Electron Microscopy
Mobile +44 (0) 7711 606967 Fax +44 (0)1280 814007
www.emcourses.com

This email is intended for the person or company named and access by anyone else is unauthorised. If you are not the person or company named, please delete this email and notify the sender.

The information in this email, including any attachments, may be confidential or legally privileged (meaning that its disclosure is protected in law). Its unauthorised disclosure, copying, distribution or use is prohibited and may be unlawful

-----Original Message-----
X-from: jacques.faerber-at-ipcms.u-strasbg.fr
[mailto:jacques.faerber-at-ipcms.u-strasbg.fr]
Sent: 07 February 2014 08:03
To: protrain-at-emcourses.com

UNSUBSCRIBE

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Larry Ackerman, Specialist
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Manager, Diabetes & Endocrinology Research Center Microscopy Core
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larry.ackerman-at-ucsf.edu

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Email: brian.miller-at-bruker-axs.com
Name: brian miller

Organization: Bruker AXS

Title-Subject: [Filtered] EM Shopping

Message: in response to Alejandro's question and several responses I have been reading, I thought it
would be good to clarify that the SEM can do nice analytical work in STEM mode and the new Bruker
detector called the Flat Quad is a great contribution to this set up as it overcomes the issue of
low signal from a TEM sample.

Although the LVEM5 from Delongh might be a very nice imaging system (I don't know but it does look
very clever), going to their website and reading the brochure it is obvious that is in no way a
system that can be used with any kind of EDS or EELs. So the analytical technique will be based
entirely on interpreting the diffraction pattern if it is obtainable.

Than you,
Brian Miller
Bruker AXS (an OEM for xray detectors on SEM and TEM)

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I was just reading the article in the January 2014 Microscopy Today issue on standardizing color in digital images using the ChromaCal microscope slide and monitor calibration system. Interesting idea but I have a question. I know the authors Barbara Foster and Jerry Sedgewick participate on the listserver so I thought I would post it here for feedback from both them and others who might have used the system.

The website for Datacolor states "The CHROMACAL slide is not suitable for use with oil immersion objectives" but Figures 1 and 4 in the Microscopy Today article were made with oil immersion objectives.

The system looks to cost around $1000 so this won't be an impulse purchase for most of us. Anyone have experience or thoughts on this?

Tom

Thomas E. Phillips, Ph.D
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)

==============================Original Headers==============================
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Email: kuryu-at-mail.rockefeller.edu
Name: Kunihiro Uryu

Organization: The Rockefeller University

Title-Subject: [Filtered] Electron Microscopist Position Available - The Rockefeller University

Message: The Rockefeller University, a premier biomedical research institution, seeks a Research
Support Specialist to join our Electron Microscopy Resource Center (EMRC).

The EMRC provides state of the art electron microscopy support for analysis of a wide variety of
biological samples, including virus, bacteria, insects, animal tissue as well as cultured cell and
isolated cellular components for structural analyses or immuno-electron microscopy. The EMRC is
equipped with two TEMs and one SEM as well as a high pressure freezing and a free substitution unit.
(http://www.rockefeller.edu/emrc/)

The Research Support Specialist will participate in all of the Electron Microscopy Resource Center's
(EMRC) daily operations, including bench work in preparation for transmission and scanning electron
microscopy and maintenance of the center. Will be responsible for all parts of sample preparation,
including cutting ultrathin sections, maintenance and operation of EM and other equipment, training
users, consulting with scientists on design of experiments, processing/analyzing data and
interpretation of results, ordering and receiving supplies, managing chemical waste compliance,
administrative support for office duties and center billing, and keeping up with the latest
methodology via familiarity with the literature. Ph.D. in biology, cell biology, bioengineering or
a related field required. Must have a minimum of 5 years of hands-on experience in electron
microscopy and have strong communication skills, and the ability to work collaboratively on a team
as well as independently on a wide variety of research projects. Must be detail-oriented, focused,
and highly motivated. Expertise in cryo- 3D is a plus.

Our culture
*inspiring, collaborative atmosphere
*strong social and environmental consciousness
*intellectually curious academic environment
*14-acre campus setting in NYC

We offer a competitive salary, comprehensive benefits, and a beautiful work environment. To apply to
this job, click the following URL, click on 'staff opportunitiesÂ’ and enter keywordÂ‘IRC11162Â’:
http://www.rockefeller.edu/hr/career.php
The Rockefeller University is an Affirmative Action/Equal Employment Opportunity/VEVRAA employer.

Kunihiro Uryu, Ph.D., Director of Electron Microscopy Resource Center

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X-from: valerie.lecomte-at-usherbrooke.ca ()

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Email: valerie.lecomte-at-usherbrooke.ca
Name: Valérie Lecomte

Organization: IOS Services Geoscientifiques

Title-Subject: [Filtered] Parts to give away Zeiss DSM940

Message: Hi everyone,
We have a few parts to give away for a SEM Zeiss DSM940(delivery at your charge)(Tungsten filament,
stage, detector, etc).
Contact me for more information!

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Email: valerie.lecomte-at-usherbrooke.ca
Name: Valérie Lecomte

Organization: IOS Services Geoscientifiques

Title-Subject: [Filtered] Parts to give away Zeiss DSM940

Message: Hi everyone,
We have a few parts to give away for a SEM Zeiss DSM940(delivery at your charge)(Tungsten filament,
stage, detector, etc).
Contact me for more information!

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Email: kuryu-at-mail.rockefeller.edu
Name: Kunihiro Uryu

Organization: The Rockefeller University

Title-Subject: [Filtered] Electron Microscopist Position Available - The Rockefeller University

Message: The Rockefeller University, a premier biomedical research institution, seeks a Research
Support Specialist to join our Electron Microscopy Resource Center (EMRC).

The EMRC provides state of the art electron microscopy support for analysis of a wide variety of
biological samples, including virus, bacteria, insects, animal tissue as well as cultured cell and
isolated cellular components for structural analyses or immuno-electron microscopy. The EMRC is
equipped with two TEMs and one SEM as well as a high pressure freezing and a free substitution unit.
(http://www.rockefeller.edu/emrc/)

The Research Support Specialist will participate in all of the Electron Microscopy Resource Center's
(EMRC) daily operations, including bench work in preparation for transmission and scanning electron
microscopy and maintenance of the center. Will be responsible for all parts of sample preparation,
including cutting ultrathin sections, maintenance and operation of EM and other equipment, training
users, consulting with scientists on design of experiments, processing/analyzing data and
interpretation of results, ordering and receiving supplies, managing chemical waste compliance,
administrative support for office duties and center billing, and keeping up with the latest
methodology via familiarity with the literature. Ph.D. in biology, cell biology, bioengineering or
a related field required. Must have a minimum of 5 years of hands-on experience in electron
microscopy and have strong communication skills, and the ability to work collaboratively on a team
as well as independently on a wide variety of research projects. Must be detail-oriented, focused,
and highly motivated. Expertise in cryo- 3D is a plus.

Our culture
*inspiring, collaborative atmosphere
*strong social and environmental consciousness
*intellectually curious academic environment
*14-acre campus setting in NYC

We offer a competitive salary, comprehensive benefits, and a beautiful work environment. To apply to
this job, click the following URL, click on 'staff opportunitiesÂ’ and enter keywordÂ‘IRC11162Â’:
http://www.rockefeller.edu/hr/career.php
The Rockefeller University is an Affirmative Action/Equal Employment Opportunity/VEVRAA employer.

Kunihiro Uryu, Ph.D., Director of Electron Microscopy Resource Center

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Hello Tom,

True, the ChromaCal slide is not suitable for oil immersion objectives.
The slide has no coverslip on the chip, and cleaning may damage the
calibration matrix. Figures 1 and 4 were taken using an oil immersion
lens for the sample, and a dry objective was used to image the ChromaCal
slide. I found I could get equivalent results regardless of the
objective used, as long as I kept the white balance the same, and set
Koehler illumination.

Having run a light microscopy lab for 15 years, the issue of color
inconsistency and inaccuracy led me to attempt making a color
calibration slide on my own. I simply could not get the filters small
enough for a microscope. Thus, my delight in working with a company
that has accomplished it at this scale. To make things even better, I
can do morphometry now using consistent color as a means to segment images.

Thanks a million for pointing out the disconnect in the article!

Jerry Sedgewick

On 2/10/2014 2:21 PM, PhillipsT-at-missouri.edu wrote:
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} I was just reading the article in the January 2014 Microscopy Today issue on standardizing color in digital images using the ChromaCal microscope slide and monitor calibration system. Interesting idea but I have a question. I know the authors Barbara Foster and Jerry Sedgewick participate on the listserver so I thought I would post it here for feedback from both them and others who might have used the system.
}
} The website for Datacolor states "The CHROMACAL slide is not suitable for use with oil immersion objectives" but Figures 1 and 4 in the Microscopy Today article were made with oil immersion objectives.
}
} The system looks to cost around $1000 so this won't be an impulse purchase for most of us. Anyone have experience or thoughts on this?
}
} Tom
}
} Thomas E. Phillips, Ph.D
} 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)
}
}
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Email: draichle-at-tampabay.rr.com
Name: Dr Erik Raichle

Organization: Raichle Diagnostic

Title-Subject: [Filtered] Specimen Size and The Limits of Lens Optics?

Message: I have 2 questions:

IÂ’ve asked this question of many people, therefore,

About what size would a specimen be when viewed through a microscope at 1000x? 70um?

Will we ever be able to view a specimen at 1000x with a field of view of 1cm and a depth of view of
1cm?

Or, would this exceed the limits of lens of optics?

Also, do we have any other technology beside lens optics that could image a 100um, or less,
specimen, such as, sonor or radar or electron beam that could be handheld?

We have identified and lost an extraordinary Mite specimen a person's skin that was less than 100um.
It was extraordinary because it looked like a spider, mites that small are wormlike. We believe it
is an unclassified pathogen and very much want to capture these specimens on peopleÂ’s skin if we can.

Or perhaps, there is a procedure that will immobilize them and isolate them under a microscope?

Anything you can think of or person who you can refer us to contact, would be most appreciated.
Thank you,
Dr Erik Raichle

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I have a customer who needs to measure the coating on 300 micron diameter
sodium sulfate core samples coated with ethyl cellulose. The core surface is
rough. Customer would like to embed in a suitable resin that will not
dissolve the ethyl cellulose coating or cause the coating to shrink/swell.
Can someone recommend an embedding media so we can pot and crossection on a
microtome for SEM ?

Thank you

Fred Hayes
Manager
Advanced Materials Characterization and Testing Lab (AMCaT)
Dept of Chemical Engineering and Material Science
3001 Ghausi Hall
Univ of CA Davis
Davis, CA 95616
530-752-0284
chms.engineering.ucdavis.edu/index.html
chms.engineering.ucdavis.edu/research/amcat/index.html

==============================Original Headers==============================
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Dr. Raichle;
To view a specimen with a 1 cm field of view at 1,000X you would need to have a computer monitor 10 meters across. So it IS possible, just difficult without a Jumbo Tron. Then you would have to stand so far back to see it, that it would get small due to being far away, so what would be the point?

John Mardinly, ASU

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Email: draichle-at-tampabay.rr.com
Name: Dr Erik Raichle

Organization: Raichle Diagnostic

Title-Subject: [Filtered] Specimen Size and The Limits of Lens Optics?

Message: I have 2 questions:

IÂ've asked this question of many people, therefore,

About what size would a specimen be when viewed through a microscope at 1000x? 70um?

Will we ever be able to view a specimen at 1000x with a field of view of 1cm and a depth of view of 1cm?

Or, would this exceed the limits of lens of optics?

Also, do we have any other technology beside lens optics that could image a 100um, or less, specimen, such as, sonor or radar or electron beam that could be handheld?

We have identified and lost an extraordinary Mite specimen a person's skin that was less than 100um.
It was extraordinary because it looked like a spider, mites that small are wormlike. We believe it is an unclassified pathogen and very much want to capture these specimens on peopleÂ's skin if we can.

Or perhaps, there is a procedure that will immobilize them and isolate them under a microscope?

Anything you can think of or person who you can refer us to contact, would be most appreciated.
Thank you,
Dr Erik Raichle

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Name: Josh Schorp

Organization: University of Missouri

Title-Subject: [Filtered] CM10 troubleshooting

Message: Upon first turning our newly renovated Philips CM10 TEM, we cannot get any of the functions
to work. The standby light on the power supply unit is illuminated as well as the ON button by the
display monitor on the electronics rack. When the ON button by the monitor is pressed, the column
makes a sort of electronic noise. Other than that there seems to be no power getting to the monitor
or any controls. Any idea on what our problem may be? Are there any controls on the power supply
that need to be turned on other than the MAINS dial?

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Dear Josh,
did you check air pressure (4 to 5 bar) and cooling water flow rate?
There are saftey switches which should be activated.

Best wishes,
Stefan

-----------------------------------------------------
Stefan Diller - Scientific Photography
Arndtstrasse 22
D - 97072 Wuerzburg Germany
++49-931-7848700 Phone
++49-931-7848701 Fax
++49-175-7177051 Mobile

Websites:
www.nanoflight.info
www.stefan-diller.com
www.electronmicroscopy.info
www.elektronenmikroskopie.info
www.zwillingsprojekt.de
www.assisi.de
Anfahrt: http://Mail.map24.com/Stefan.Diller
-----------------------------------------------------

Am 12.02.14 01:52, schrieb microscopylistserver-noreply-at-microscopy.com:
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}
} Title-Subject: [Filtered] CM10 troubleshooting
}
} Message: Upon first turning our newly renovated Philips CM10 TEM, we cannot get any of the functions
} to work. The standby light on the power supply unit is illuminated as well as the ON button by the
} display monitor on the electronics rack. When the ON button by the monitor is pressed, the column
} makes a sort of electronic noise. Other than that there seems to be no power getting to the monitor
} or any controls. Any idea on what our problem may be? Are there any controls on the power supply
} that need to be turned on other than the MAINS dial?
}
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7, 22 -- From stefan.diller-at-t-online.de Wed Feb 12 00:44:20 2014
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Dear all,

We are currently searching for two SEMs in particular one equipped with a focussed ion beam column and mass spectrometry capabilities. Are any of you aware of manufacturers and does anyone have experience with such instruments?

The formal request for information is available in this link - https://www.ungm.org/Public/Notice/25524

Thanks a lot,
Ernesto

Mr Ernesto CHINEA-CANO | Laboratory Assistant |
Office of Safeguards Analytical Services | Environmental Sample Laboratory | Department of Safeguards |
International Atomic Energy Agency | IAEA Laboratories, Reaktorstrasse 1, A-2444 Seibersdorf, Austria|
Email: E.Chinea-Cano-at-iaea.org | T: (+43-1) 2600-28583 | F: (+43-1) 2600-28577|
Follow us on www.iaea.org

This email message is intended only for the use of the named recipient. Information contained in this email message and its attachments may be privileged, confidential and protected from disclosure. If you are not the intended recipient, please do not read, copy, use or disclose this communication to others. Also please notify the sender by replying to this message and then delete it from your system.

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Hi all

Following my e-mail in answer to Alejandro Hinojos's question, I was
contacted by the DeLonghi representative. So I want just to add some
precisions to my former comments.

First, I want to point out that I didn't gave any feedback about the
quality of the results that come out of this instrument. To be right and
fair, I must say that the tests which were made for us gave pictures in
the same quality range than with the STEM device for SEM. As we didn't
perform the test ourself, I cannot say anything about the ease of use of
the instrument and so on.

Secondly our final choice has ben dictated by to main raisons :
-first we have a FE-SEM
-second we didn't have the monney for the DeLonghi intrument and no hope
to get it in a near future. But we had enough for the STEM device, to be
fitted to the existing FE-SEM.
The global result is less expend and more flexibility, but less
troughput possibility and no self-service use.

It is evident that all situations are not the same. A lab which has no
SEM or TEM can have benefit to spend 150 kE in a table-tob TEM/SEM for a
fast and easy to use instrument. Or a big EM service may have an
interest too, as a tool usefull to sort TEM grids before going to the
high end TEM.

I hope that these comments will clarify the contexte of my former mail.
My goal was only to point out an other possibility as Alejandro Hinojos
said they have soon SEM and TEM in their lab.

Best Regards

J. Faerber
IPCMS-DSI
Institut de Physique et Chimie des Matériaux de Strasbourg
Département Surfaces et Interfaces
23, rue de Loess ; BP43
67034 Strasbourg CEDEX 2
France

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

Le 07/02/2014 09:12, jacques.faerber-at-ipcms.u-strasbg.fr a écrit :
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} Hi
}
} A collegue was interested by this instrument last year and made some
} tests. His goal was to have an easy access to a TEM like instrument, to
} make fast controls in nanoparticules syntesis (organics with heavy
} metals), as the acess to our TEMs need some days of delay.
}
} Hearing about the price of that instrument, I proposed him to test too a
} STEM device to be fitted on our FE-SEM.
} Comparing prices and performences (principally the versatility), the
} choice was easy. The STEM device for the SEM gives better images, the
} possibility to choose the HV up to 30 keV and a price 8x less expensive...
}
} The real adavantage of this table-top TEM I could see is that it can be
} used easy by non-microcopists, like table-top SEMs, with on the other
} hand the limitations of the undestanding such people will have of what
} they do.
}
} So if you soon have a (FE)SEM, it could be usefull to consider if the
} STEM device for SEM would not be a better solution. But I'm interested
} on what your tests will bring you as conclusions.
}
} Hope it helps
}
} J. Faerber
} IPCMS-DSI
} Institut de Physique et Chimie des Matériaux de Strasbourg
} Département Surfaces et Interfaces
} 23, rue de Loess ; BP43
} 67034 Strasbourg CEDEX 2
} France
}
} Tel 00 33(0)3 88 10 71 01
} Fax 00 33(0)3 88 10 72 48
} E-mail : Jacques.Faerber-at-ipcms.unistra.fr
}
} Le 07/02/2014 03:28, ahinojos2-at-miners.utep.edu a écrit :
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} } Hello Microscopists,
} } Here at the University of Texas at El Paso we are searching for a new EM to purchase. We mainly use Hitachi SEM and TEM for Hard materials characterization. We are mainly searching for an EM with specialization in polymers characterization. In our search we stumbled upon the LEVM5 Benchtop TEM from Delong America for polymers. The unit claims to have TEM, SEM, and STEM capabilities. We here in the southwest were wondering what are/were your experiences the LVEM5 or other Delong EM’s? We were also wondering how are the products from the Delong America Company in the perspective of materials characterization? If anybody wouldn't mind sharing those experiences with us that would be great.
} }
} }
} } Thanks for your thoughts and answers in advance
} } Alejandro Hinojos
} }
} }
} }
} }
} } ==============================Original Headers==============================
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Josh,

First, is the "Panel Dim" knob pushed in? Turns on the panel. "Data Dim"
has to be up to be able to see anything on the monitor.
There is a delay while the computer runs self tests.
Power supply, left half:
The 4 boxes in the top row should all have green lights on when the
'scope is on, regardless of whether or not the panel is on.
Same for the 30V "dbl" box in the middle row, and the large black box in
the bottom.
When the HT button is pushed, the 24V filament box green light will come
on in the middle row, as well as the left-most box in that row (no label
on our CM-10).
There should be no red lights - check the 2 right-most boxes in the
middle row. Those are safeties, and have reset buttons. If those are
showing red lights, turn off the HT and push the reset buttons.

But. The column shouldn't be making any noise at all. Is it really
electronic, or is it a crackly water-flowing noise? Could be air in the
cooling lines, which is likely if the 'scope was torn apart. The water
lines would need to be disconnected at the exits, and water run through
to remove any air in the lines.
Were the water lines cleaned with H2O2 or CLR (brand) cleaner? If not,
they may be blocked by crud or corrosion. 1 liter 3% H2O2 in the chiller
tank for 2 hr - overnight, then 4 or 5 flushes with clean water should
clear them.

If it is an electronic noise ... check the connections. If they're OK,
then arg. Problem. Get out the sacrificial rooster or undergrad and ...

Phil

} Email: jcsmtf-at-mail.missouri.edu
} Name: Josh Schorp
}
} Organization: University of Missouri
}
} Title-Subject: [Filtered] CM10 troubleshooting
}
} Message: Upon first turning our newly renovated Philips CM10 TEM, we cannot get any of the functions
} to work. The standby light on the power supply unit is illuminated as well as the ON button by the
} display monitor on the electronics rack. When the ON button by the monitor is pressed, the column
} makes a sort of electronic noise. Other than that there seems to be no power getting to the monitor
} or any controls. Any idea on what our problem may be? Are there any controls on the power supply
} that need to be turned on other than the MAINS dial?
--
Philip Oshel
Microscopy Facility Supervisor
Biology Department
024C Brooks Hall
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576

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The Biozentrum of the University of Basel is one of the leading institutes worldwide for molecular and biomedical basic research and teaching. It is home to more than 30 research groups with scientists from over 40 countries. Research at the Biozentrum focuses on the areas of Cell Growth & Development, Infection Biology, Neurobiology, Structural Biology & Biophysics and Computational & Systems Biology. With its more than 550 employees, the Biozentrum is the largest department at the University of Basel’s Faculty of Science.

The University of Basel maintains two sites for electron microscopy, the C-CINA and the ZMB. The Center for Cellular Imaging and NanoAnalytics (C-CINA) performs advanced electron microscopy research in life sciences, see http://c-cina.org. The ZMB provides TEM and SEM services to the University, see http://zmb.unibas.ch. The C-CINA and the ZMB together operate 15 electron microscopes including a FEI Titan Krios with a Gatan K2 Summit direct electron detector, FEI Polara, CM200FEG, Helios FIB-SEM and a Quanta200FEG with a Gatan 3View.

We have an opening for an

=========================
EM Service Engineer
=========================

Your responsibilities will include the maintenance and repair of all electron microscopes of C-CINA and the ZMB. We do not have any factory maintenance contracts but may request additional help from the factory when needed. Responsibilities of this position are maintenance and repair of the electron microscopes including; problem diagnosis, ordering replacement parts from the factory, and repair. This includes: the Titan Krios Autoloader, cryo-stages, FEG and high voltage supplies, vacuum systems, mechanics, electronics and software backups.

In addition, upon interest, this engineer may participate in electron microscope hardware development projects, which involve hardware design, fabrication in collaboration with our mechanical workshop, construction, and development of electronic control in collaboration with an electronic workshop. Participation in research projects concerning structural biology data collection with these instruments is also possible. C-CINA is a University research group; we develop instruments and methods, and apply them to study the high-resolution 3D structure of biological specimens.

Salary and living conditions in Basel are attractive. The lab language is English.

Your profile
Experience in EM service for TEM, ideally also for SEM and FIB. Hardware repair experience.

We offer
This is a full-time position permanently funded by the Rektorat of the University of Basel. A highly sought after position in one of the most beautiful countries in Europe. We offer a welcoming working environment in an international and multidisciplinary group covering biochemistry, electron microscopy, structural biology, and data analysis. We are an equal opportunity employer.

Basel is an international city with people from 150 nations. Located at the border where three countries meet - Switzerland-Germany-France. It is Europe’s most important life sciences hub. Basel provides a high standard of living and a rich and varied cultural atmosphere.

For further information, please contact (ABSOLUTE CONFIDENTIALITY IS ASSURED):

Henning Stahlberg, Director
Center for Cellular Imaging and NanoAnalytics (C-CINA)
Prof. for Structural Biology and Biophysics, Biozentrum,
WRO-1058, Mattenstrasse 26
University of Basel, CH-4058 Basel, Switzerland
Tel: +41-61-387 32 62 ; http://c-cina.org
mailto:Henning.Stahlberg-at-unibas.ch

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Email: mamiller-at-coh.org
Name: Marcia M. Miller

Organization: Beckman Research Institute, City of Hope

Title-Subject: [Filtered] Using agar to secure cell pellets

Message: The agar that we are currently using to secure cell pellets is causing a really annoying
background all over the sections both inside and outside of cells. This is a terrible graininess
that we have seen now in several preps. What we have is bacterial grand from Applichem. We are
pretty sure this is the cause of the graininess. Can anyone recommend another agar or another means
of securing cell pellets? Sometimes users of our core can provide only small quantities of cells.
Thanks in advance for suggestions. Best wishes, Marcia

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Hi Marcia:

We have been using the low-melting-point agarose (NOT agar). It works great. You can find it at Sigma (Cat# A9414).

Zhaojie

Zhaojie Zhang, Ph. D.
Director, Jenkins Microscopy Facility
University of Wyoming
Laramie, WY 82071
PHONE: 307-766-3038
FAX: 307-766-5625

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Email: mamiller-at-coh.org
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Organization: Beckman Research Institute, City of Hope

Title-Subject: [Filtered] Using agar to secure cell pellets

Message: The agar that we are currently using to secure cell pellets is causing a really annoying background all over the sections both inside and outside of cells. This is a terrible graininess that we have seen now in several preps. What we have is bacterial grand from Applichem. We are pretty sure this is the cause of the graininess. Can anyone recommend another agar or another means of securing cell pellets? Sometimes users of our core can provide only small quantities of cells.
Thanks in advance for suggestions. Best wishes, Marcia

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Marcia,
We have encase small fragile objects between Formvar films on
a wire loop. Although this does require appropriate obeissance to the
Formvar Spirits, the outcome in terms of structure preservation is
excellent. You can read about it in Wu et al. 2012 Nature Protocols
7: 1113- 1124. (I can send you a pdf off line if you are interested).
That paper describes using the method on a small plant root but there
is no reason in principle why it would not work for a cell pellet.
Probably you won't want to switch horses like this but sending a long
the info in case you are interested. Good luck!
Tobias

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} Email: mamiller-at-coh.org
} Name: Marcia M. Miller
}
} Organization: Beckman Research Institute, City of Hope
}
} Title-Subject: [Filtered] Using agar to secure cell pellets
}
} Message: The agar that we are currently using to secure cell pellets
} is causing a really annoying
} background all over the sections both inside and outside of cells.
} This is a terrible graininess
} that we have seen now in several preps. What we have is bacterial
} grand from Applichem. We are
} pretty sure this is the cause of the graininess. Can anyone
} recommend another agar or another means
} of securing cell pellets? Sometimes users of our core can provide
} only small quantities of cells.
} Thanks in advance for suggestions. Best wishes, Marcia
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Hi Erin:

We use 1.5% agarose in PBS (I assume other medium works too). We first mix the agarose with PBS in an eppendorf tube, then immerse the tube in boiling water for a few minutes. The agarose would be completed 'melted' at this point. We then transfer the tube to a 37 degree C water bath, to let it cool down to 37C. The agarose (melting point 36C) will stay melted at 37C.

We then mixed 1 part of cells and 1 part of agarose at 37C, then bring the mixture to room temperature. The mixture will become solid right away (no need for ice)!

Hope this helps.

Zhaojie

X-from: Erin Tranfield [mailto:etranfield-at-igc.gulbenkian.pt]
Sent: Thursday, February 13, 2014 8:57 AM
To: Z.J. Zhang

Hi Marcia:

We have been using the low-melting-point agarose (NOT agar). It works great. You can find it at Sigma (Cat# A9414).

Zhaojie

Zhaojie Zhang, Ph. D.
Director, Jenkins Microscopy Facility
University of Wyoming
Laramie, WY 82071
PHONE: 307-766-3038
FAX: 307-766-5625

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Dear All,
here are some images from a Raspberry leaf:
http://www.electronmicroscopy.info/sem_images/index.htm?1
Can someone please help me identify the structures seen in the mid and right image?
Especially those with the small "spheres" on the surface?
Is this mostly mold on the surface?

Best wishes,
Stefan

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Arndtstrasse 22
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Websites:
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www.zwillingsprojekt.de
www.assisi.de
Anfahrt: http://Mail.map24.com/Stefan.Diller
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realname - Anne Howson
Email - AKHPhoto-at-pacbell.net
ORGANIZATION - Richardson Bay Audubon Center
EDUCATION - Graduate College
LOCATION - Tiburon, Ca.
QUESTION - We have an Olympus SZ61 Microscope with an Hitachi CCD color
camera (model kP-D20BU attached which we connect to a projector to teach
or docents and trainees about plankton in San Francisco Bay. Several
months ago the system stopped projecting color images and will now only
project black & white. This is not nearly as effective and dramatic.
Can you suggest what might be wrong? We tried buying a new projector,
but that did not fix the problem. Thanks for any help you can give us.
As a small non-profit educational facility, we do not have much $.

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You should be able to get excellent in-person help from the San
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Hello All,
Please, could anybody give us an advice or hint on following problem?
We have a trouble with our Philips CM100. I can see the light on main
screen but cannot focus the beam with Intensity button. At both end
positions of Intensity button range I can hear the beeps. Microscope
restart did not help.
Without any touching the Intensity button C2 Condenser current in LM
mode is oscillating between 3900 mA and 3924 mA. In M mode it is
oscillating from 3868 mA to 3944 mA.
C1 Condenser currents seem to be OK (389, 424, 470, 550, 640, 748,
855,1026, 1266, 1486, 2166 mA for spotsize from 1 to 11).

Thanking you in advance.

Best regards
Oldrich

--
Oldrich Benada
Institute of Microbiology AS CR. v.v.i.
Videnska 1083
142 20 Prague 4
Czech Republic

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realname - David Glick
Email - glickdavidb-at-gmail.com
ORGANIZATION - San Joaquin Delta College
EDUCATION - Undergraduate College
LOCATION - Stockton Ca. USA
SUBJECT_OF_QUESTION - Level of Education
QUESTION - I am currently working towards a two year certificate in EM
and considering continuing my education on a part-time basis. My
question is two part: First; If I am considering working in the
materials/crystalline sector how far can I expect to get with just a
certificate? Second; Does obtaining a degree above a bachelors improve
upward mobility in the industry? The Reason I ask these questions is
that I am a 45 year old re-entry student and do not want to needlessly
spend time and money in a university if it won't improve my employment
standard.

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A tough question to answer.

In my 40 years of chemistry and microscopy I've seen changes. Today industry hires BS to do the work technician did 30 years ago. 60 years ago you might have gotten an interview for a position as a technician but now you don't get in the door with out your sheep skin. I took my BS degree in Chemistry and scrambled to take microscopy courses at McCrone and other places. I found a position as a Microscopist and never looked back.

I worked for a man who had a PhD in the metallography of welding, He was moving up the management chain powdered by his drive and the PhD.
So yes, I would say a degree is better than no degree. An advanced degree is better than a BS, but pick your field with care. Add other skill sets, like programming, electronics to round out your skill set.

Nothing is guarantied. I have a friend with a BS in chemical engineering and a masters in chemistry. He's in law school because he couldn't find a job in those fields. But he will not leave California either.......

If it was a perfect world, you could try to find a position which would allow you to finish school while employed. My limited experience is you then need to change job as soon as possible because management will always see you as a technician.

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realname - David Glick
Email - glickdavidb-at-gmail.com
ORGANIZATION - San Joaquin Delta College
EDUCATION - Undergraduate College
LOCATION - Stockton Ca. USA
SUBJECT_OF_QUESTION - Level of Education
QUESTION - I am currently working towards a two year certificate in EM
and considering continuing my education on a part-time basis. My
question is two part: First; If I am considering working in the
materials/crystalline sector how far can I expect to get with just a
certificate? Second; Does obtaining a degree above a bachelors improve
upward mobility in the industry? The Reason I ask these questions is
that I am a 45 year old re-entry student and do not want to needlessly
spend time and money in a university if it won't improve my employment
standard.

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I am trying to hone my understanding of diffraction in microscopy: particularly the slit diffraction analogy and how diffraction affects resolution in microscopy.
1. If you take a sub-resolution point source of light (GFP molecule) and image it through an objective, you get a central maximum and a series of concentric disks due to diffraction during image formation. Question- What is the source of the diffraction? There are no slit like openings that are less than the wavelength of the light. I am presuming this diffraction due light diffracting at the edges of the objective? (I could not find a good written/diagrammatic description of the issue- can someone point me to one?)

2. The separation of Airy disk central maxima as an explanation for resolution makes some intuitive sense, but is only taking account of the NA of the objective as if there are no other sources of diffraction other than the objective. This would be reasonable for a purified fluorescent molecule/protein attached to a coverslip. However, in cells, you now add many other sources of diffraction as light passes into and out of a cell. So would it be more correct to say that the calculated resolution is a maximal or optimal resolution? I am presuming that this is routinely not acheived in cell/tissue imaging? Do we have an estimate of how much resolution is lost by a molecule being in the middle of a cell (ie. what you get from a single superresolution activated molecule in PALM) as compared to a purified molecules in isolation? Presumably you can back calculate this from the actual diffraction pattern measured for single activated molecules in a cell vs. in isolation?
Thanks- Dave

David Knecht, Ph.D.
Professor and Head of Core Microscopy Facility
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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Â
Hi David,

1) Actually the Youngâ€™s slit experiment doesnâ€™t require the width or the distance between the slits to be less than the wavelength of light. You can do it using a slide that youâ€™ve put a thin black coating on (using model airplane paint for example) and then scratch it twice with a razor and shine a light through it. If you do your trigonometry, youâ€™ll find out that if you put the slits really close (as in a few hundred microns or even a mm) you can see the diffraction pattern pretty clearly some feet away. In a microscope, you have apertures, and tubes bounding the light path and that is what is causing your diffraction. If you are operating at high NA then often the limiting aperture is the objective. Youâ€™ll get diffraction whenever the light path from one side of the aperture is on the order of one wavelength of light different than in the middle of the aperture, practically speaking, high mag.

2) Point resolution limits described by airy disks etc are really rules of thumb. As you note, the resolution limit is the smallest feature you can separate from your image, nothing more or less. Usually that is much less than the Airy resolution due to inhomogeneities in the sample, like you describe. On the other hand, theoretically, if you know EXACTLY the diffraction characteristics of your entire system, and your sample is composed of point sources, and you have infinite signal to noise, you can separate two different points that are infinitely close together. You can make a quick demonstration with Matlab or a similar program by plotting two Gaussians with FWHM of 100 units, and centroids separated by 1 unit, and then do a fit to extract the separation between the two centroids. You wonâ€™t have any trouble even though this scenario is resolving peaks far, far below the "resolution limit." As soon as you add even a little noise, you cannot make the fit work. As soon as you allow for any uncertainty in the FWHM or skewness of the peaks, then you also canâ€™t get a good number from your fit. I encourage you to try this, actually, it is very informative just how little noise or uncertainty in the peak shapes is required to destroy your ability to resolve the two peaks, and if you play with it for a while will wind up deriving a practical resolution limit â€œlawâ€ of your own. Chances are you will wind up with a limit not far from the FWHM of the Gaussians. If you like programming you can do this with Airy disks and youâ€™ll â€œreinventâ€ the Airy resolution limit. In a real microscope, you always have some aberrations, and your sample always has some inhomogeneities (otherwise why are you looking at it?). So the Airy resolution is a very practical limit and very useful, but there is no deep fundamental reason why the grand master of the universe says thatâ€™s the resolution limit. Itâ€™s just a good demarcation that was chosen based on a reasonable application of diffraction through an aperture.

I hope this helps!

Zack Gainsforth
Space Sciences Laboratory, UC Berkeley
7 Gauss Way
Berkeley, CA 94720
cell: 626-437-9186(tel://cell:%20626-437-9186)
zackg-at-ssl.berkeley.edu(mailto:zackg-at-ssl.berkeley.edu)

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Reply: david.knecht-at-uconn.edu(mailto:david.knecht-at-uconn.edu) david.knecht-at-uconn.edu(mailto:david.knecht-at-uconn.edu)

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Title-Subject: [Filtered] CM10 Error Message

Message: Upon turning on our CM10 for the first time after the install there is a message saying
"Pneumatics". I can't seem to get the ODP to turn on, and twice now the viewing glass has been
pushed out by the compressed air. Does anyone have any ideas on what the problem may be?

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Title-Subject: [Filtered] CM10 HIVAC

Message: I have not been able to activate the HIVAC and UHV on our CM10. The pressures of the pironi
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to get the P2 down below 13.3?

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Hi Josh,

A "pneumatics" error message generally means that your air pressure is
too low to operate the valves of the microscope. The FEI TEMs generally
want to have ~6 bar air pressure.

If your viewing window is being forced out, it sounds like you may have
a more serious problem than the pneumatics error. The viewing chamber
is not supposed to have a positive pressure but be under vacuum. Do you
have a pressurized air supply hooked to the camera vent valve (valve V12
on the vacuum schematic)? It's possible that you have a leaky valve.

Cheers,
Henk

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Hi Josh,
The ODP needs a heat up time of about 20 min. after the vacuum system has been off for a while. P3 = 0 means it is 100+. When the ODP is heated the vacuum system will continu by opening the valve between the ODP and the camera, then P3 will go down as well.
Hans Janssen
Netherlands Cancer Institute (NKI/AvL)
Amsterdam

_________________________________
Van: microscopylistserver-noreply-at-microscopy.com [microscopylistserver-noreply-at-microscopy.com]
Verzonden: vrijdag 14 februari 2014 23:57
Aan: Hans Janssen
Onderwerp: [Microscopy] viaWWW:CM10 HIVAC

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Email: jcsmtf-at-mail.missouri.edu
Name: Josh Schorp

Organization: University of Missouri

Title-Subject: [Filtered] CM10 HIVAC

Message: I have not been able to activate the HIVAC and UHV on our CM10. The pressures of the pironi
gauges are: P1=82, P2=19, P3=0, IGP=0. This is after the pre-vavc has been running for roughly 20
minutes. Once the pre-vac is turned off P2 goes to about ~70. Is it normal to take longer than this
to get the P2 down below 13.3?

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Name: Mr Richard Robert White

Organization: UVA

Title-Subject: [Filtered] 2000FX double tilt holder

Message: We are in need of the plug on a double tilt holder which plugs into a JEOL 2000FX TEM. If
you have a broken double tilt holder, we could use the cable and plug from the holder.

Thanks,

Richard White
University of Virginia
rrw3q-at-virginia.edu

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Josh,

The "Pneumatics" error typically means your air pressure from the
compresser to the valves is too low. Ours reads ~90 psi normally.
Do you have any of the manuals? The appendix in the operator's manuals
has the error codes.
The ODP not coming on generally means insufficient cooling water, or the
cooling water is too hot/too cold. It should be ~20-22 deg C (~70 F, if
your Haskris cooler has the usual US temp gauge).

The viewing glass being pushed out means your vent pressure is high.
Your column/camera/specimen airlock vent is hooked up to a dry nitrogen
tank, yes? That should be at 1-2 psi, no more.

Re: your other email about pressures:
I have not been able to activate the HIVAC and UHV on our CM10. The
pressures of the pironi gauges are: P1=82, P2=19, P3=0, IGP=0. This is
after the pre-vavc has been running for roughly 20 minutes. Once the
pre-vac is turned off P2 goes to about ~70. Is it normal to take longer
than this to get the P2 down below 13.3?

Our values in normal operation, after pumping overnight:
P1 = 37
P2 = 74
P3 = 34
IGP = 14

Your P1 is high, which may mean a leak. If the column was taken apart,
the leak is probably in one of those seals, and may not be visible under
inspection. Clean.
I've never seen a P2 value as low as yours. Seems wrong, likely
connected with a leak, but I'd like a service engineer to explain it.
P3 reads 0 either because the pressure is too high for it to come on, or
the pressure is less than 34 - P3 goes to zero when the pressure drops
below 34.

Also the P2 and/or P3 gauges may be malfunctioning. I'd assume a leak to
start with. That's the usual bugaboo after taking apart an EM column.

Do you have an ion getter pump on your CM-10? If yes, disconnect it and
*don't* run it until you get the rest of the vacuum system working
(unplug connector X8, back of the column just below the IGP).

Phil

} Email: jcsmtf-at-mail.missouri.edu
} Name: Josh Schorp
}
} Organization: University of Missouri
}
} Title-Subject: [Filtered] CM10 Error Message
}
} Message: Upon turning on our CM10 for the first time after the install there is a message saying
} "Pneumatics". I can't seem to get the ODP to turn on, and twice now the viewing glass has been
} pushed out by the compressed air. Does anyone have any ideas on what the problem may be?

--
Philip Oshel
Microscopy Facility Supervisor
Biology Department
024C Brooks Hall
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576

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realname - M Raja
Email - plspce-at-amararaja.co.in
ORGANIZATION - Amararaja Batteries Ltd
EDUCATION - Undergraduate College
LOCATION - Tirupathi, Andhra pradesh, India
SUBJECT_OF_QUESTION - Color metallography-reg
QUESTION - how to prepare etchent for lead base alloys and hardware &
software requirement and also difficulties / limitations of color
metallographic analysis. Actually i need details about benefits of color
micrograph.
Please revert back in above mail as soon as possible. Thank you in
advance for your support.

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I'd try "Metallogtaphy: Principles and Practices by George F Vander Voot

It's an excellent book.

In some color staining systems, as I understand it, the physical orientation of the metal crystal results in a coloration that is constant for that orientation in that specific sample. Color photomicroscopy lets you image where these orientations are. Color provides contrast that simple gray scale may not.

-----Original Message-----
X-from: oshel1pe-at-cmich.edu [mailto:oshel1pe-at-cmich.edu]
Sent: Monday, February 17, 2014 8:49 AM
To: Frank Karl

***************************************************************************************
Forwarded from "Ask a Microscopist"
Please remember that the person asking the question is likely
not a member the listserver, and
**any reply should go directly to the poster**
as well as to the list.
Using the "reply" function in your email does *not* send your answer
to the person asking the question.
Please copy their email address from their question.
****************************************************************************************

realname - M Raja
Email - plspce-at-amararaja.co.in
ORGANIZATION - Amararaja Batteries Ltd
EDUCATION - Undergraduate College
LOCATION - Tirupathi, Andhra pradesh, India
SUBJECT_OF_QUESTION - Color metallography-reg
QUESTION - how to prepare etchent for lead base alloys and hardware &
software requirement and also difficulties / limitations of color
metallographic analysis. Actually i need details about benefits of color
micrograph.
Please revert back in above mail as soon as possible. Thank you in
advance for your support.

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This email and any of its attachments may contain confidential information intended only for the use of the addressee(s). If the reader of this email is not the intended recipient or the employee or agent responsible for delivering it to the intended recipient, you are hereby notified that any dissemination or copying of this email is strictly prohibited. If you have received this email in error, please notify us by return email at info-at-ardl.com, permanently delete the email, and destroy any printouts. If this email contains test data and/or draft reports, you are hereby notified that only a signed original test report will contain official results, a copy of which resides in the project folder located at ARDL, Inc. Thank you. Akron Rubber Development Laboratory, Inc.

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We've just updated our image processing instructions here at the NUANCE Center from Northwestern University.

These instructions are super easy to use and walk the user step-by-(sometimes painful) step to do all sorts of normal image processing procedures one might normally come across, from adjusting levels, to calculations, to some totally cheating techniques of drawing a mag line marker, to several different procedures to apply false color to an image.

THE EXTRA SPECIAL PART is the chapter on what I call Multi-Detector Color. Now I know I did not invent this technique, BUT I've worked out some pretty simple procedures that will allow you to make these really fantastic color images, even if you only have 1 SE detector in your SEM.
It's totally free, so please check it out and let us know if you have corrections or suggestions.

http://www.nuance.northwestern.edu/docs/epic-pdf/Basic_Photoshop_for_Electron_Microscopy_2014.pdf

ERiC Jay Miller
Microscopy & Imaging Specialist
Electron Probe Instrumentation Center

Northwestern University
Mail: 2036 Cook Hall
Office: 1152 Cook Hall
2220 Campus Drive
Evanston, IL 60208-3108

ph: (847) 467-0789
fax: (847) 467-657

http://www.nuance.northwestern.edu

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I would appreciate hearing suggestions for a good mounting technique for
imaging activated carbon and synthetic diamond particles (1-200 micron).
When using standard conductive double sided adhesive carbon tabs on stubs,
it is obviously quite challenging to get adequate contrast between the
particles and the background (carbon tab).

Likewise, in doing particle size analysis it helps to not have touching
particles and clumps. Any techniques that have been found to be successful
would be interesting to hear.

Thank You!!

Mike Toalson
Western Analytical Solutions, LLC
o • 208.899.4425 m • 208.340.3895
w • www.materialanalyzers.com e • mike-at-materialanalyzers.com

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Does anybody know of software for building Montage images for a Titan TEM? We would like to build a large image from a multitude of high resolution plant cell images.

Pete Eschbach
Oregon State University

Sent from my iPhone

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I use ImageJ for that, not sure if it's suited for your type of images
though.

Nick Botto
Microprobe Specialist
UC Davis Earth and Planetary Sciences Microprobe Lab
Website: http://microprobe.geology.ucdavis.edu/index.html
Calendar: http://microprobe.geology.ucdavis.edu/calendar/probe.htm
ph. 530-752-6582

On 2/18/14, 2:29 PM, peter.eschbach-at-comcast.net wrote:
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} Does anybody know of software for building Montage images for a Titan TEM? We would like to build a large image from a multitude of high resolution plant cell images.
}
} Pete Eschbach
} Oregon State University
}
} Sent from my iPhone
}
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The ImageJ plug-in MosaicJ is pretty easy to use and allows big images. I
believe you need the TurboReg plug-in installed as well, as it can
automatically stitch as long as you put them down close to the right
position. I've used it successfully several times.
Good Luck!
Larry Scipioni
ZS Genetics

-----Original Message-----
X-from: peter.eschbach-at-comcast.net [mailto:peter.eschbach-at-comcast.net]
Sent: Tuesday, February 18, 2014 5:30 PM
To: LES-at-ZSGENETICS.COM

Does anybody know of software for building Montage images for a Titan TEM?
We would like to build a large image from a multitude of high resolution
plant cell images.

Pete Eschbach
Oregon State University

Sent from my iPhone

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I've used the GIMP for this application with a lot of success. Something
else to look at would be this:
http://research.microsoft.com/en-us/um/redmond/groups/ivm/ICE/

-Jacob

On 14-02-18 02:29 PM, peter.eschbach-at-comcast.net wrote:
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} Does anybody know of software for building Montage images for a Titan TEM? We would like to build a large image from a multitude of high resolution plant cell images.
}
} Pete Eschbach
} Oregon State University
}
} Sent from my iPhone
}
} ==============================Original Headers==============================
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Hi Pete,

I have used the commercial program Corel Photopaint (part of the Coreldraw suite of programs) extensively over the years for TEM and other montaging.
It is manual montaging, but it handles any file size (I have done up to montage around 300 GB), and it has a feature where the selected image is subtracted from what is under it, meaning you can quickly see which parts of the image are aligned because they go completely black.

The programs others have mentioned are also good options.

Kind regards
Duane Harland
AgResearch
New Zealand

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Does anybody know of software for building Montage images for a Titan TEM? We would like to build a large image from a multitude of high resolution plant cell images.

Pete Eschbach
Oregon State University

Sent from my iPhone

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Hello,

lead etching you found in the book:

"Metallographic etchnig" by Gûnter Petzow
edited by ASM 1999,
black white photo lead structure was published in the practicall
metalography many years ago (1970 ?)

best regards

best regards

Chris Hûbner

Cracow University of Technology
Faculty of Mechanical Engineering

----- Original Message -----
X-from: {oshel1pe-at-cmich.edu}
To: {hubner-at-mech.pk.edu.pl}
Sent: Monday, February 17, 2014 2:50 PM

Dear Microscopists,

maybe someone can give me a hint on how to proceed now.
We just encountered a problem with our Tecnai Osiris in
STEM mode:
We reduced the acceleration voltage from 200kV to 80kV
to deal with a little bit sensitive samples. In TEM Mode
we didnÂ´t had any problems, but in STEM we see a distortion.
I uploaded a picture* of the problem.
We suspect some magnetic field could be responsible for this
behavior.

What do you think? And what could we do to find the reason?

Thank you in advance!

best regards,

Max

* https://www.dropbox.com/s/1ued2rl94y81rfo/19-02-2014-80kV_HAADF.jpg

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Mike,
I'm going to assume you have a powder.

I start with double sticky conductive tape on a stub. I spread the powder out on a glass slide and blow the powder off the slide and onto the stub. Just a puff, a little breath, a mouse fart. A small mouse.

You can position the stub below or across from your slide. Too much powder on stub? Blow it off, use a bigger mouse this time.

What you want is a single layer of crystals/particles, separated as much as possible from each other. The stub will look sample deficient to the eye, check it with a stereomicroscope and I suspect you'll find plenty.

I coat these samples with Au or Pt, twice. One coating straight on and then I lay the SEM mount in the chamber at an angle to get the "underside" of the particles. Works great. I've been running organic accelerators for rubber this way for years and have always been happy with results.

Good Luck!

Frank

-----Original Message-----
X-from: mike-at-materialanalyzers.com [mailto:mike-at-materialanalyzers.com]
Sent: Tuesday, February 18, 2014 4:03 PM
To: Frank Karl

I would appreciate hearing suggestions for a good mounting technique for
imaging activated carbon and synthetic diamond particles (1-200 micron).
When using standard conductive double sided adhesive carbon tabs on stubs,
it is obviously quite challenging to get adequate contrast between the
particles and the background (carbon tab).

Likewise, in doing particle size analysis it helps to not have touching
particles and clumps. Any techniques that have been found to be successful
would be interesting to hear.

Thank You!!

Mike Toalson
Western Analytical Solutions, LLC
o * 208.899.4425 m * 208.340.3895
w * www.materialanalyzers.com e * mike-at-materialanalyzers.com

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1. "Metallographic etching", 2nd by Gûnter Petzow , ASM 1999
Or "Metallographic etching", 1st by Gûnter Petzow , ASM 1978

Have about 2-3 pages each with several comments, but no photomicrographs.

2. For a good discussion and B&W photomicrographs see:

Allen, Metallographic Tech for Lead-Acid Battery Plates, Microstructural
Science, Vol 6, 31-43, 1978

3. For coloring lead phases try the Copper Alloy etch set out by:

Beraha: Color Metallography. ASM 1977.

4. Coloring can be an advantage if it adds to the contrast desired to
detect or demonstrate a particular phase. This will depend on the material
at hand.

5. M Raja, please report back how it turns out.

Tony

……………………………………………………………………………………………………………
Andrew Anthony “Tony” Havics, CIH, PE
Environmental, Health & Safety, Microscopy, Materials Science & Forensic
Engineering
pH2, LLC
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-----Original Message-----
X-from: hubner-at-mech.pk.edu.pl [mailto:hubner-at-mech.pk.edu.pl]
Sent: Wednesday, February 19, 2014 7:19 AM
To: ph2-at-sprynet.com

Hello,

lead etching you found in the book:

"Metallographic etchnig" by Gûnter Petzow
edited by ASM 1999,
black white photo lead structure was published in the practicall
metalography many years ago (1970 ?)

best regards

best regards

Chris Hûbner

Cracow University of Technology
Faculty of Mechanical Engineering

----- Original Message -----
X-from: {oshel1pe-at-cmich.edu}
To: {hubner-at-mech.pk.edu.pl}
Sent: Monday, February 17, 2014 2:50 PM

Hi

Beckert - Klemm "Handbuch der metallographischen Ätzverfahren"
VEB Deutsch. Verlag f. Grundstoffindustrie - 1962

Sorry, it's in german, but very usefull. Always on my shelf !

There are 10 pages with different solutions vor lead and lead alloys,
with 2 BW pictures and an (old now) bibliography.

Hope it helps

J. Faerber
IPCMS-DSI
Institut de Physique et Chimie des Matériaux de Strasbourg
Département Surfaces et Interfaces
23, rue de Loess ; BP43
67034 Strasbourg CEDEX 2
France

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

Le 19/02/2014 13:25, hubner-at-mech.pk.edu.pl a écrit :
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} Hello,
}
} lead etching you found in the book:
}
} "Metallographic etchnig" by Gûnter Petzow
} edited by ASM 1999,
} black white photo lead structure was published in the practicall
} metalography many years ago (1970 ?)
}
} best regards
}
} best regards
}
} Chris Hûbner
}
} Cracow University of Technology
} Faculty of Mechanical Engineering
}
}
}
}
}
}
} ----- Original Message -----
} X-from: {oshel1pe-at-cmich.edu}
} To: {hubner-at-mech.pk.edu.pl}
} Sent: Monday, February 17, 2014 2:50 PM
} Subject: [Microscopy] Ask-A-Microscopist: etchant for lead alloys & any
} advantage to color
}
}
} }
} }
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} }
} } realname - M Raja
} } Email - plspce-at-amararaja.co.in
} } ORGANIZATION - Amararaja Batteries Ltd
} } EDUCATION - Undergraduate College
} } LOCATION - Tirupathi, Andhra pradesh, India
} } SUBJECT_OF_QUESTION - Color metallography-reg
} } QUESTION - how to prepare etchent for lead base alloys and hardware &
} } software requirement and also difficulties / limitations of color
} } metallographic analysis. Actually i need details about benefits of color
} } micrograph.
} } Please revert back in above mail as soon as possible. Thank you in
} } advance for your support.
} }
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Hi Peter,

} Does anybody know of software for building Montage images for a Titan TEM? We would like to build a large image from a multitude of high resolution plant cell images.

a number of options to stitch existing images has been discussed already - if you are interested in software that drives automated acquisition to build montages, have a look at SerialEM (actually a tomography software):

http://bio3d.colorado.edu/SerialEM/index.html

It works on the Titan platform and can also do the stitching.

Best,

Guenter

--
Dr. Guenter Resch
Nexperion e.U. - Solutions for Electron Microscopy - www.nexperion.net
Mattiellistrasse 3/17, 1040 Vienna, Austria - Phone +43 664 94 17 210
Registered at Commercial Court Vienna, FN 397677w - VAT Reg. ATU67962234

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The Montage building software you use will be dependent not so much on the microscope but how the images are named and in what order they are collected. If you are using a FEI Eagle camera, you will need to convert the proprietary image format to .tif or .jpg then proceed. GIMP does this well. We use an AMT camera and the images are automatically named according to their x-y position in the montage, for example 0006R1C6 refers to an image in the first row and the sixth column. How the stage moves from one image to the next during collection is essential information. We have used AdobePhotoshop for image montages, but it is slow compared to FIJI and may not complete large montages. For montages up to 19x19 images, I would suggest using FIJI. For image sets of this size you will also want a 64 bit operating system with exceptional graphics capabilities. Download Fiji (http://fiji.sc/wiki/index.php/Downloads). Fiji is a distribution of ImageJ together with Java, Java 3D and several plugins organized to assist research in life sciences, targeting image registration, stitching, segmentation, feature extraction and 3D visualization, among others. Using FIJI you can direct the program to the folder in which the images are stored and also define the naming nomenclature so that the montage software will find each successive image. There is quite a bit of flexibility built into FIJI as to how the program finds the images; we have had the most success with the position defined by the file name. Once you download FIJI, bring up the screen where images in this format can be loaded (FIJI/Stitching/Deprecated/Stitch grid of images). I would be happy to send a word document with specific instructions with screen shots on how to work with images in this format to anyone who might find it useful.

Cheers,

Doug

Douglas R. Keene
Assistant Investigator
Micro-Imaging Center
Shriners Hospital for Children
3101 SW Sam Jackson Park Road
Portland, Oregon 97239
Office: 503-221-3434
Mobile: 503-819-3600

} Does anybody know of software for building Montage images for a Titan TEM? We would like to build a large image from a multitude of high resolution plant cell images.
}
} Pete Eschbach
} Oregon State University
}
} Sent from my iPhone
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MSA Listserver -

There are many montaging programs.
I can suggest PTGUI which some
of our SEM users have used for
montages with hundreds of images.
It is not picky on the filenames, filetypes,
and/or file order.

PTGUI stands for Panorama Tools (Graphic User Interface).

regards,

- Jim

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I did not send any of you a message in the last two days. Please do not open if you did receive one for I do not know where the message originated.
-- Pat
Patricia Stranen Connelly
NHLBI EM Core Facility
National Institutes of Health
Building 14E Room 111B
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301-496-3491

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3, 58 -- To: ChuckCharles A Garber {cgarber-at-2spi.com} ,
3, 58 -- DanDan Connelly
3, 58 -- {dansconnelly-at-hotmail.com} ,
3, 58 -- "Sherman, Debra" {dsherman-at-purdue.edu} ,
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Deal Listers,

My mail has been hacked or whatever you you call it. I am attempting to find which email was broken into but this was the fastest way to get to a lot of you. I have not sent any messages to the Listerv earlier today.

-- Pat
Patricia Stranen Connelly
NHLBI EM Core Facility
National Institutes of Health
Building 14E Room 111B
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301-496-3491

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Email: alpinto-at-fm.ul.pt
Name: Andreia Pinto

Organization: Instituto de Medicina Molecular

Title-Subject: [Filtered] JEOL JEM-100CX II diaphragms

Message: We have here at the Institute an old Jeol JEM-100CX II that was disconnected for many
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options in acquiring/buying diaphragms for this brand/model, can you help me??
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Thank you very much!!

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Hi Max,
You're probably seeing the effect of a stray AC field on your scan. A
paper by Muller et al. from Ultramicroscopy describes ways to design a
room to minimize this kind of distortion:
http://www.sciencedirect.com/science/article/pii/S0304399106001033

Do you notice the distortion change when you change your scan rate?
Right now, it appears that the period of the field's oscillation is
equal to the time necessary to complete a couple of scan lines. If you
can find a scan rate at which the distortion period is equal to the time
necessary to complete an integer number of scan lines, you should be
able to calculate the frequency of the distortion (I bet it's 60 Hz!)
which may help you locate the source of the field.

Tyler

On 2014-02-19 6:50, raven2020-at-gmx.de wrote:
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} Dear Microscopists,
}
} maybe someone can give me a hint on how to proceed now.
} We just encountered a problem with our Tecnai Osiris in
} STEM mode:
} We reduced the acceleration voltage from 200kV to 80kV
} to deal with a little bit sensitive samples. In TEM Mode
} we didnÃ‚Â´t had any problems, but in STEM we see a distortion.
} I uploaded a picture* of the problem.
} We suspect some magnetic field could be responsible for this
} behavior.
}
} What do you think? And what could we do to find the reason?
}
} Thank you in advance!
}
} best regards,
}
} Max
}
}
} * https://www.dropbox.com/s/1ued2rl94y81rfo/19-02-2014-80kV_HAADF.jpg
}
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Good evening everyone,
I am have some problem with my formvar grids. till 6 months ago they
were all good but now all (i even checked new one from sealed box) of
them are extremely fragile under beam (they tear apart even while
going from low mag to high mag mode). Not only that, it seems they are
developing some sort of static charge as beam shifts can be seen
inside microscope when i insert those grids. here is attached pics of
how a beam edge (a shadow of objective aperture) is behaving near a
hole (made during changing mode) in formvar substrate. Any ideas what
am doing wrong?

https://sites.google.com/site/auxilliarylinks/

Regards
Amit Gupta
Research Scholar
Dept. of Inorganic and Physical Chemistry, IISc

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PhD scholarship in Electron microscopy of electrochemical processes in liquids

for more information see
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and

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Imbibition

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RJ Perz-Edwards- I also suspected the same problem hence cleaned the
holder properly, though did not check the top part. Well most of grids
are little vulnerable during that transition but now they are almost
sure to break apart. also if u do stem you can see them expanding and
breaking. It reders them almost useless.
shashi singh- no we are buying it. Glow discharge is not available at
the moment to try but i did try grounding them while making sample...
did not help. Will try ethanol shortly.
Ross- Although manufacturer is same but they are copper only, we can
see it in EDS. Besides they were working perfectly before, how come
all of sudden?

Regards
Amit Gupta
Research Scholar
Prof. S.Vasudevan's lab
Dept. of Inorganic and Physical Chemistry, IISc

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Dear Listservers,
I am planning an HPF hybrid (pre-light chemical fixation) technique on attached cells, for HM-20 post embed IEM. My plan is to lightly fix the cells for 1 hr (4% PF 0.5% Ga 0.1M HEPES with 5mM MgCl2), rinse in 0.1M HEPES 3.5% sucrose with 5mM MgCl2 (243 mOsmols), then scrape and pellet the cells. I will resuspend the pellet in 2% agarose (in the cells media minus serum) just before HPF. My question is has anyone used HEPES as an IEM buffer, and at what concentration (molarity)? I have read that a slightly hypotonic solution is better for IEM, I believe the cultured cells are isotonic at 300 mOsmols. Also for IEM, is there really a difference between 4% PF and 2% PF? Any suggestions are welcome.

sincerely,
Michael Delannoy

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Michael - I use HEPES routinely in my regular fix and for immunocytochemistry. I typically use 70 mM NaCl, 30 mM HEPES, 2 mM CaCl2, pH 7.4. I based my choice of osmolarity on the old half-strength Karnovsky's buffer which if I remember was 100 mM cacodylate. You see a lot of people referring to half-strength Karnovsky's as being ~1000 mOsm (or full-strength as being 2010 mOsm). These numbers assume the formaldehyde and glutaraldehyde contribute to the osmolarity of the fix. They might count in an osmometer measurement but if something crosses the membrane it doesn't count to the cell. My buffer is clearly "hypo-osmotic" to a mammalian cell at ~300 mOsm but empirically it seems to cause the least shrinkage or swelling. Your results may vary. Somebody once told me that Karnovsky picked a high aldehyde concentration to show they didn't contribute to the osmolarity but I don't know if that is true.

Is there a difference between 2% PF and 4% PF? Yes, I believe 4% has twice as much PF! Sorry, I couldn't resist the joke. I know exactly what you mean. I haven't seen much difference for cell lines and almost always use 2%. I use 2% PF by itself based on my use of "half-strength" Karnovsky's. The old "full-strength Karnovsky's fix was 4% PF and 5% glutaraldehyde - half-strength cut the aldehyde concentrations in half but not the buffer concentration. I suspect with an excess of aldehyde, the same amount of crosslinking occurs if you wait long enough.

Good luck. Tom

Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
phillipst-at-missouri.edu

http://biology.missouri.edu/people/?person=88
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Dear Listservers,
I am planning an HPF hybrid (pre-light chemical fixation) technique on attached cells, for HM-20 post embed IEM. My plan is to lightly fix the cells for 1 hr (4% PF 0.5% Ga 0.1M HEPES with 5mM MgCl2), rinse in 0.1M HEPES 3.5% sucrose with 5mM MgCl2 (243 mOsmols), then scrape and pellet the cells. I will resuspend the pellet in 2% agarose (in the cells media minus serum) just before HPF. My question is has anyone used HEPES as an IEM buffer, and at what concentration (molarity)? I have read that a slightly hypotonic solution is better for IEM, I believe the cultured cells are isotonic at 300 mOsmols. Also for IEM, is there really a difference between 4% PF and 2% PF? Any suggestions are welcome.

sincerely,
Michael Delannoy

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Dear Michael,
dear all,

I thank Prof. Phillips for the thorough explanation of his opinion on the matter.
I also would like to second use of 2%PFA (FA made from paraformaldehyde powder right before)instead of 4% for cell cultures/cultured cells.
I have read papers stating 30 min. initial fixation ( -at-room temperature ) would be sufficient for cultured cells.
Needless to say it would be of advantage to block unreacted aldehydes after fixation by washes in your buffer + 50mM NH4Cl
(ROTH J et al, Enhancement of Structural Preservation and Immunohistochemical Staining in low temperature embedded pancreatic tissue, JHC 29,663-671(1981)

I have seen also an interesting post (on ResearchGate, www.researchgate.net,
Thread: http://www.researchgate.net/topic/Histology/post/Alternatives_to_formalin_fixation
Re by Thomas Kroneis . Medizinische Universität Graz = Med. Univ. Graz, Austria) on using HOPE(R) as an alternative for FA-fixation in IHC.
HOPE. HOPE = Hepes-glutamic acid buffer mediated Organic solvent Protection Effect
T. Kroneis wrote in his post :

{ { ....Additionally to what has been posted above: there is another fixation around: HOPE.
HOPE = Hepes-glutamic acid buffer mediated Organic solvent Protection Effect
(German: http://dcs-diagnostics.de; http://dcs-diagnostics.de/index.php?sprache=de ;
ENGL: http://dcs-diagnostics.de/index.php?sprache=en ),

HOPE(R)-Fixation:
GERM: http://dcs-diagnostics.de/index.php?sprache=de&&qid=48 ,
ENGL: http://dcs-diagnostics.de/index.php?sprache=en&&qid=63 ).
This fixation yields as good morphological quality as FFPE material and circumvents the problem with the antibodies not recognizing the epitopes! Now the bad news: You need to place your sample directly into the pre-cooled HOPE I solution, second, it takes you quite some time (fixation 16h - 72h plus another day). - So if there is just no possibility to have it snap frozen, maybe it would be possible to have the respective solution stored there (crucial: at +2°C!). If you do not perform RNA analysis afterwards (do you?), you also might think of having the sample just stored in a tube and transferred to your site (in case of HOPE - just cooling, do not use buffers etc.). Maybe you should also think of getting solved the logical issue. Good science needs perfect conditions (you know: shit in - shit out!) Hope this helped, contact me, if you need further assistance (our lab protocol, ...)} } ,
and, same thread:
Re by: Sebastian Marwitz . Research Center Borstel
{ { The probably best described alternative to PFA is the HOPE-technique:
(Olert et al.,Pathol Res Pract. 2001;197(12):823-6); Wiedorn et al., Pathol Res Pract. 2002;198(11):735-40; Kähler et al., J Histochem Cytochem. 2010 Mar;58(3):221-8; Marwitz et al., J Histochem Cytochem. 2011 Jun;59(6):601-14.; Vollmer et al., Rom J Morphol Embryol. 2006;47(1):15-9; Goldmann et al., Pathol Res Pract. 2005;201(8-9):599-602.; Goldmann et al., Am J Pathol. 2003 Dec;163(6):2638-40) .
It is a non-crosslinking fixation with acetone dehydration und subsequent paraffin-embedding. There is compared to PFA only minimal denaturation of nucleic acids and allows a multitude of read-outs from one tissue/cell block (qPCR, proteomics, RNA in situ hybridization, microarrays, ...). Most important, you don´t need any antigen-retrieval and are able to dilute your primary antibodies very high. If you are skilled in handling PFA-fixed specimen, there will be no big difference in tissue processing and cutting. End of citation} }

Also I have a pdf in my files, written by T.W. BRIMBLE: The developing Role of the Zwitterionic Buffer (1981) wherein use of HEPES buffer in various combinations or molar concentrations in Cell/organ culture, bacterial culture, blood proteins, cryogenic tissue storage and chemical data will be presented and discussed. Since at the beginning of the "era of zwitterionic buffers" there is only a small subtitle "EM" showing interesting effects in fixation of plant material (total 48 references, file= 5,5MB).
So if anyone is interested, I could send that file to any requester..

Have a nice and warm weekend,
especially in the US,

best regards,
Wolfgang

Wolfgang MUSS
EM-Lab
Univ.Inst.Pathology
SALK-LKH(Gen. Hosp.) and PMU
(Private Paracelsus Medical University SALZBURG)
SALZBURG-AUSTRIA
DISCLAIMER: no affiliation, or financial interest in any of the companies or their products.

} -----Ursprüngliche Nachricht-----
} Von: PhillipsT-at-missouri.edu [mailto:PhillipsT-at-missouri.edu]
} Gesendet: Freitag, 21. Februar 2014 00:25
} An: Muß Wolfgang
} Betreff: [Microscopy] RE: HEPES and HPF
}
}
}
}
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} (Re: PhillipsT-at-missouri.edu)
} Michael - I use HEPES routinely in my regular fix and for
} immunocytochemistry. I typically use 70 mM NaCl, 30 mM HEPES, 2 mM
} CaCl2, pH 7.4. I based my choice of osmolarity on the old half-strength
} Karnovsky's buffer which if I remember was 100 mM cacodylate. You see a
} lot of people referring to half-strength Karnovsky's as being ~1000
} mOsm (or full-strength as being 2010 mOsm). These numbers assume the
} formaldehyde and glutaraldehyde contribute to the osmolarity of the
} fix. They might count in an osmometer measurement but if something
} crosses the membrane it doesn't count to the cell. My buffer is clearly
} "hypo-osmotic" to a mammalian cell at ~300 mOsm but empirically it
} seems to cause the least shrinkage or swelling. Your results may vary.
} Somebody once told me that Karnovsky picked a high aldehyde
} concentration to show they didn't contribute to the osmolarity but I
} don't know if that is true.
}
} Is there a difference between 2% PF and 4% PF? Yes, I believe 4% has
} twice as much PF! Sorry, I couldn't resist the joke. I know exactly
} what you mean. I haven't seen much difference for cell lines and almost
} always use 2%. I use 2% PF by itself based on my use of "half-strength"
} Karnovsky's. The old "full-strength Karnovsky's fix was 4% PF and 5%
} glutaraldehyde - half-strength cut the aldehyde concentrations in half
} but not the buffer concentration. I suspect with an excess of aldehyde,
} the same amount of crosslinking occurs if you wait long enough.
}
} Good luck. Tom
}
}
} Thomas E. Phillips, Ph.D
} Professor of Biological Sciences
} Director, Molecular Cytology Core
} 2 Tucker Hall
} University of Missouri
} Columbia, MO 65211-7400
} 573-882-4712 (office)
} 573-882-0123 (fax)
} phillipst-at-missouri.edu
}
} http://biology.missouri.edu/people/?person=88
} http://www.biotech.missouri.edu/mcc/
}
} -----Original Message-----
} X-from: delannoy-at-jhmi.edu [mailto:delannoy-at-jhmi.edu]
} Sent: Thursday, February 20, 2014 12:42 PM
} To: Phillips, Thomas E.
} Subject: [Microscopy] HEPES and HPF
}
}
}
}
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}
} Dear Listservers,
} I am planning an HPF hybrid (pre-light chemical fixation) technique on
} attached cells, for HM-20 post embed IEM. My plan is to lightly fix
} the cells for 1 hr (4% PF 0.5% Ga 0.1M HEPES with 5mM MgCl2), rinse in
} 0.1M HEPES 3.5% sucrose with 5mM MgCl2 (243 mOsmols), then scrape and
} pellet the cells. I will resuspend the pellet in 2% agarose (in the
} cells media minus serum) just before HPF. My question is has anyone
} used HEPES as an IEM buffer, and at what concentration (molarity)? I
} have read that a slightly hypotonic solution is better for IEM, I
} believe the cultured cells are isotonic at 300 mOsmols. Also for IEM,
} is there really a difference between 4% PF and 2% PF? Any suggestions
} are welcome.
}
} sincerely,
} Michael Delannoy
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Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

A while ago while working on the histology of naturally distended rabbit urinary bladders that had been subjected to outlet obstruction, I used a (P)HCHO at 4% in a buffer (probably phosphate) at 4 degrees C as a 'quick' fix (10 minutes) of the whole, distended, bladder followed by removal of the urine and replacement with more cold buffered (P)HCHO to further fix the mucosal side of the bladder wall. The point here is that the 10 minute immersion 'froze' the smooth muscle so that the bladder did not contract when removing the urine and replacing with fixative.

This method worked as well on muscle that had been used in physiologic contraction studies to preclude contraction when the tissue was stimulated/studied in Hank's Balanced Salt Solution rather than in traditional phosphate or Tyrode buffer. The problem is that Hank's is not approved via general use, but if you want your muscle to look like muscle when you are done, something like Hank's is helpful.

As to the concentration of HCHO, it is my belief that all histologic protocols support/require the use of excesses of formaldehyde, and everything else. This is an edgy topic, and likely not worth too much time. The key issue with all methods of tissue preservation lies with the necessity to avoid having to provide an explanation for why one's tissue looks so bad!!! What IS/WAS liver must LOOK like liver!

Cheers, and hope this starts arguments,

Fred Monson.

What else does one do when one gets old other than get cranky?

Frederick C. Monson, PhD
Technical Director
Center for Microanalysis and Imaging, Research and Training (CMIRT)
West Chester University of Pennsylvania
Schmucker Science South - Room SSS-024
MailDrop: Geology-Astronomy
750 South Church Street
West Chester, PA, 19383
610-738-0437
fmonson-at-wcupa.edu

HomePage: http://cmirt.wcupa.edu (with link to instrument Scheduler)

-----Original Message-----
X-from: W.Muss-at-salk.at [mailto:W.Muss-at-salk.at]
Sent: Friday, February 21, 2014 4:49 AM
To: Monson, Frederick

Dear Michael,
dear all,

I thank Prof. Phillips for the thorough explanation of his opinion on the matter.
I also would like to second use of 2%PFA (FA made from paraformaldehyde powder right before)instead of 4% for cell cultures/cultured cells.
I have read papers stating 30 min. initial fixation ( -at-room temperature ) would be sufficient for cultured cells.
Needless to say it would be of advantage to block unreacted aldehydes after fixation by washes in your buffer + 50mM NH4Cl
(ROTH J et al, Enhancement of Structural Preservation and Immunohistochemical Staining in low temperature embedded pancreatic tissue, JHC 29,663-671(1981)

I have seen also an interesting post (on ResearchGate, www.researchgate.net,
Thread: http://www.researchgate.net/topic/Histology/post/Alternatives_to_formalin_fixation
Re by Thomas Kroneis . Medizinische Universität Graz = Med. Univ. Graz, Austria) on using HOPE(R) as an alternative for FA-fixation in IHC.
HOPE. HOPE = Hepes-glutamic acid buffer mediated Organic solvent Protection Effect T. Kroneis wrote in his post :

{ { ....Additionally to what has been posted above: there is another fixation around: HOPE.
HOPE = Hepes-glutamic acid buffer mediated Organic solvent Protection Effect
(German: http://dcs-diagnostics.de; http://dcs-diagnostics.de/index.php?sprache=de ;
ENGL: http://dcs-diagnostics.de/index.php?sprache=en ),

HOPE(R)-Fixation:
GERM: http://dcs-diagnostics.de/index.php?sprache=de&&qid=48 ,
ENGL: http://dcs-diagnostics.de/index.php?sprache=en&&qid=63 ).
This fixation yields as good morphological quality as FFPE material and circumvents the problem with the antibodies not recognizing the epitopes! Now the bad news: You need to place your sample directly into the pre-cooled HOPE I solution, second, it takes you quite some time (fixation 16h - 72h plus another day). - So if there is just no possibility to have it snap frozen, maybe it would be possible to have the respective solution stored there (crucial: at +2°C!). If you do not perform RNA analysis afterwards (do you?), you also might think of having the sample just stored in a tube and transferred to your site (in case of HOPE - just cooling, do not use buffers etc.). Maybe you should also think of getting solved the logical issue. Good science needs perfect conditions (you know: shit in - shit out!) Hope this helped, contact me, if you need further assistance (our lab protocol, ...)} } , and, same thread:
Re by: Sebastian Marwitz . Research Center Borstel { { The probably best described alternative to PFA is the HOPE-technique:
(Olert et al.,Pathol Res Pract. 2001;197(12):823-6); Wiedorn et al., Pathol Res Pract. 2002;198(11):735-40; Kähler et al., J Histochem Cytochem. 2010 Mar;58(3):221-8; Marwitz et al., J Histochem Cytochem. 2011 Jun;59(6):601-14.; Vollmer et al., Rom J Morphol Embryol. 2006;47(1):15-9; Goldmann et al., Pathol Res Pract. 2005;201(8-9):599-602.; Goldmann et al., Am J Pathol. 2003 Dec;163(6):2638-40) .
It is a non-crosslinking fixation with acetone dehydration und subsequent paraffin-embedding. There is compared to PFA only minimal denaturation of nucleic acids and allows a multitude of read-outs from one tissue/cell block (qPCR, proteomics, RNA in situ hybridization, microarrays, ...). Most important, you don´t need any antigen-retrieval and are able to dilute your primary antibodies very high. If you are skilled in handling PFA-fixed specimen, there will be no big difference in tissue processing and cutting. End of citation} }

Also I have a pdf in my files, written by T.W. BRIMBLE: The developing Role of the Zwitterionic Buffer (1981) wherein use of HEPES buffer in various combinations or molar concentrations in Cell/organ culture, bacterial culture, blood proteins, cryogenic tissue storage and chemical data will be presented and discussed. Since at the beginning of the "era of zwitterionic buffers" there is only a small subtitle "EM" showing interesting effects in fixation of plant material (total 48 references, file= 5,5MB).
So if anyone is interested, I could send that file to any requester..

Have a nice and warm weekend,
especially in the US,

best regards,
Wolfgang

Wolfgang MUSS
EM-Lab
Univ.Inst.Pathology
SALK-LKH(Gen. Hosp.) and PMU
(Private Paracelsus Medical University SALZBURG) SALZBURG-AUSTRIA
DISCLAIMER: no affiliation, or financial interest in any of the companies or their products.

} -----Ursprüngliche Nachricht-----
} Von: PhillipsT-at-missouri.edu [mailto:PhillipsT-at-missouri.edu]
} Gesendet: Freitag, 21. Februar 2014 00:25
} An: Muß Wolfgang
} Betreff: [Microscopy] RE: HEPES and HPF
}
}
}
}
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} (Re: PhillipsT-at-missouri.edu)
} Michael - I use HEPES routinely in my regular fix and for
} immunocytochemistry. I typically use 70 mM NaCl, 30 mM HEPES, 2 mM
} CaCl2, pH 7.4. I based my choice of osmolarity on the old
} half-strength Karnovsky's buffer which if I remember was 100 mM
} cacodylate. You see a lot of people referring to half-strength
} Karnovsky's as being ~1000 mOsm (or full-strength as being 2010
} mOsm). These numbers assume the formaldehyde and glutaraldehyde
} contribute to the osmolarity of the fix. They might count in an
} osmometer measurement but if something crosses the membrane it doesn't
} count to the cell. My buffer is clearly "hypo-osmotic" to a mammalian
} cell at ~300 mOsm but empirically it seems to cause the least shrinkage or swelling. Your results may vary.
} Somebody once told me that Karnovsky picked a high aldehyde
} concentration to show they didn't contribute to the osmolarity but I
} don't know if that is true.
}
} Is there a difference between 2% PF and 4% PF? Yes, I believe 4% has
} twice as much PF! Sorry, I couldn't resist the joke. I know exactly
} what you mean. I haven't seen much difference for cell lines and
} almost always use 2%. I use 2% PF by itself based on my use of "half-strength"
} Karnovsky's. The old "full-strength Karnovsky's fix was 4% PF and 5%
} glutaraldehyde - half-strength cut the aldehyde concentrations in half
} but not the buffer concentration. I suspect with an excess of
} aldehyde, the same amount of crosslinking occurs if you wait long enough.
}
} Good luck. Tom
}
}
} Thomas E. Phillips, Ph.D
} Professor of Biological Sciences
} Director, Molecular Cytology Core
} 2 Tucker Hall
} University of Missouri
} Columbia, MO 65211-7400
} 573-882-4712 (office)
} 573-882-0123 (fax)
} phillipst-at-missouri.edu
}
} http://biology.missouri.edu/people/?person=88
} http://www.biotech.missouri.edu/mcc/
}
} -----Original Message-----
} X-from: delannoy-at-jhmi.edu [mailto:delannoy-at-jhmi.edu]
} Sent: Thursday, February 20, 2014 12:42 PM
} To: Phillips, Thomas E.
} Subject: [Microscopy] HEPES and HPF
}
}
}
}
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} Dear Listservers,
} I am planning an HPF hybrid (pre-light chemical fixation) technique on
} attached cells, for HM-20 post embed IEM. My plan is to lightly fix
} the cells for 1 hr (4% PF 0.5% Ga 0.1M HEPES with 5mM MgCl2), rinse in
} 0.1M HEPES 3.5% sucrose with 5mM MgCl2 (243 mOsmols), then scrape and
} pellet the cells. I will resuspend the pellet in 2% agarose (in the
} cells media minus serum) just before HPF. My question is has anyone
} used HEPES as an IEM buffer, and at what concentration (molarity)? I
} have read that a slightly hypotonic solution is better for IEM, I
} believe the cultured cells are isotonic at 300 mOsmols. Also for IEM,
} is there really a difference between 4% PF and 2% PF? Any suggestions
} are welcome.
}
} sincerely,
} Michael Delannoy
}
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From lytcinhu-at-natm.ru Sat Feb 22 04:37:38 2014
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Email: wul-at-mail.montclair.edu
Name: Laying Wu

Organization: education

Title-Subject: [Filtered] Microscopy training workshop 2014

Message: Dear Listservers,

Montclair State University and Hitachi High Technologies Inc. are pleased to announce a low-cost,
one-day Light and Electron Microscopy workshop for new and intermediate-level microscopists. The
workshop will take place on Friday, March 21, 2014 at Montclair State University, Montclair New
Jersey. The workshop will introduce participants to a wide range of the techniques associated with
light microscopy, electron microscopy, atomic force microscopy and sample preparation. Guest
presenters will join us from Hitachi High Technologies of America, Bruker AXS Inc., and Hitech
Instruments Inc. Students, faculty, researchers, and industry users working in biology, geology,
environmental science and material science are encouraged to apply.

Please visit the workshop website for further details and registration instructions:
http://www.montclair.edu/csam/mmrl/workshop/

Montclair State University is located in Montclair, NJ. We are reachable via the New Jersey Transit
train system (Montclair Heights and Montclair State University stations) and by bus. Please visit us
at www.montclair.edu.

with best wishes,

Laying Wu

wul-at-mail.montclair.edu
TEL: 973-655-2028

Dr. Laying Wu
Director, Microscopy and Microanalysis Research Laboratory
Montclair State University
1 Normal Ave, Montclair, NJ 07043
USA

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Email: doug.skinner-at-bruker-nano.com
Name: Doug Skinner

Organization: Bruker Nano Analytics

Title-Subject: [Filtered] Job Opportunity at Bruker - Applications Specialist

Message: Bruker Nano Analytics, a leading manufacturer of Analytical Instrumentation, has a job
opening for an Applications Specialist in their Billerica, MA Microanalysis Lab. This division
specializes in EDS, WDS, micro-XRF and EBSD on electron microscopes.

The Applications Specialist position is a Sales, Service and Customer support position. The
responsibilities are to assist customers, demonstrate the microanalysis instruments and to provide
technical sales support for the Microanalysis Product Line.

Primary Responsibilities:
* Demonstrate equipment at Bruker and customer sites
* Analysis of customer samples
* Customer support on-site and via phone/e-mail
* User training of the Bruker Microanalysis System
* Produce appropriate materials for customer training
* Preparation of technical sales materials
* Assist Service in resolving customer problems
* Perform other tasks as assigned by manager

This position involves up to 50% travel, primarily in North America and Latin America.

Candidates interested in this position require a minimum of a BachelorÂ’s degree in a Physical
Science discipline with a degree in Chemistry, Geology or Materials Science preferred. Two years
SEM/X-ray microanalysis and EBSD experience, including sample preparation, is preferred.

Requirements include excellent communication skills and proficiency with Microsoft Office.
Language: English fluent; preferably Spanish as a second language.

Interested candidates should complete an online application at: www.bruker.com

Bruker Nano Analytics offers a competitive salary and comprehensive benefits package including
health and dental insurance, Flexible Spending Account, company sponsored life and disability
insurance, 401(k) plan with company matching components, and vacation and sick days.

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Dear Colleagues,
We would like to bring your attention to the call for abstracts:
https://www.avs.org/Meetings-Exhibits/Call-For-Abstracts
to the AVS 61st International Symposium & Exhibition (AVS-61),
which will be held November 9-14, 2014 in Baltimore, Maryland.
It is a good opportunity to explore and report on new developments
in microscopy techniques applied to new materials, devices and interfaces.
In addition, on the behalf of the organizers, we invite you to submit an abstract
before the deadline of Monday, May 5th, to the special topical session
"Novel Trends in Synchrotron and FEL-Based Analysis"
https://www.avs.org/Meetings-Exhibits/Call-For-Abstracts/Focus-Topics#Synchrotron.
This focus session is organized for the first time and aims at dissemination of
the unique research opportunities, offered by the advanced experimental techniques
at X-ray, VUV or IR synchrotron and free electron laser facilities, to investigate
the properties of matter with unprecedented space, spectral and time resolution.
One or two more invited presentation will be selected by the program committee
evaluating the abstracts. We look forward to receiving your abstract and seeing
you November in Baltimore, Maryland!
_______________________________________________________________________________
Andrei Kolmakov, PhD
Project Leader
Center for Nanoscale Science and Technology
National Institute of Standards and Technology

NIST 100 Bureau Drive
Bldg. 216/Rm.B117
Gaithersburg, MD 20899-6204
Phone: (301) 975-4724
Fax: (301) 975-2303
E-mail: andrei.kolmakov-at-nist.gov
URL: http://www.nist.gov/cnst/kolmakov.cfm

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3, 31 -- From: "Kolmakov, Andrei A." {andrei.kolmakov-at-nist.gov}
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3, 31 -- Subject: AVS 61st International Symposium & Exhibition (AVS-61) and focus
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Folks -
I have two Gatan 600 ion-mills that
are up for grabs on Long Island, NY.
Both are not 100% working, put
contain 90% of parts. Together,
you can easily make a working unit.
You would have to come here yourself
and load (or crate) them yourself.

If nobody offers to take them in their entirety,
then anyone is welcome to come here to
strip them for parts. I am out that business.

Either way, they are being surplussed in one month.

Next, does anyone have a PDF copy of the
instruction manual and schematics for a Gatan 691
ion-mill?
thank you and regards,
- Jim

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4, 26 -- Subject: Gatan 600 ion-mills up for grabs and need manual for 691
4, 26 -- From: Jim Quinn {jquinn11733-at-gmail.com}
4, 26 -- To: Microscopy-at-microscopy.com
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Folks -
I have two Gatan 600 ion-mills that
are up for grabs on Long Island, NY.
Both are not 100% working, put
contain 90% of parts. Together,
you can easily make a working unit.
You would have to come here yourself
and load (or crate) them yourself.

If nobody offers to take them in their entirety,
then anyone is welcome to come here to
strip them for parts. I am out that business.

Either way, they are being surplussed in one month.

Next, does anyone have a PDF copy of the
instruction manual and schematics for a Gatan 691
ion-mill?
thank you and regards,
- Jim

==============================Original Headers==============================
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4, 26 -- Message-ID: {CAO_SM8Kk_7H504-iMRzp+2-eY5tcQZXje=LD6JCp2cneKFNBZQ-at-mail.gmail.com}
4, 26 -- Subject: Gatan 600 ion-mills up for grabs and need manual for 691
4, 26 -- From: Jim Quinn {jquinn11733-at-gmail.com}
4, 26 -- To: Microscopy-at-microscopy.com
4, 26 -- Content-Type: text/plain; charset=ISO-8859-1
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Email: jcsmtf-at-mail.missouri.edu
Name: Josh Schorp

Organization: University of Missouri

Title-Subject: [Filtered] CM10 pneumatics

Message: I am receiving a Pneumatics message on the screen of our CM10 and I believe this to be the
reason while none of the vacuums will start. The air line on our machine runs first to V11, but it
has not been allowed past V11. Is there a reason for this? I'm not sure how this valve works, I took
it off and it seems to be just a metal unit with no moving parts. I'm guessing the black electrical
piece that connects to it has something to do with its operations. Does anyone have any insight on
how to open this valve? I am running ~90 psi to it.

Login Host: 161.130.188.19
Listserver Email Form V - 20120416
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12, 28 -- To: MicroscopyListServer-Forward {microscopy-at-microscopy.com}
12, 28 -- Subject: viaWWW:CM10 pneumatics
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Folks -
Those two ion-mills are spoken for.
Also, I now have the manual for the 691.
Thank you Justin and Mike!
regards,
- Jim

} Folks -
} I have two Gatan 600 ion-mills that
} are up for grabs on Long Island, NY.
} Both are not 100% working, put
} contain 90% of parts. Together,
} you can easily make a working unit.
} You would have to come here yourself
} and load (or crate) them yourself.
} If nobody offers to take them in their entirety,
} then anyone is welcome to come here to
} strip them for parts. I am out that business.
} Either way, they are being surplussed in one month.
} Next, does anyone have a PDF copy of the
} instruction manual and schematics for a Gatan 691
} ion-mill?
} thank you and regards,
} - Jim

==============================Original Headers==============================
3, 26 -- From jquinn11733-at-gmail.com Sun Feb 23 14:28:46 2014
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3, 26 -- Subject: ion-mills are now spoken for
3, 26 -- From: Jim Quinn {jquinn11733-at-gmail.com}
3, 26 -- To: Microscopy-at-microscopy.com
3, 26 -- Content-Type: text/plain; charset=ISO-8859-1
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Hi Josh,

The microscope air supply should *NOT* be attached to V11. V11 is the
microscope vent valve and is used to release the vacuum in the column
and camera chamber. It should never have more than a fraction of a psi
supplied to it. Higher pressure can force the viewing chamber window
out with great force and cause serious injury to anyone nearby! Venting
the column with high pressure air when the sample rod is inserted can
also blow the sample rod out with some force and damage it.

The air for the valves is supplied through the "Watts" regulator to the
distribution block on the rear of the microscope column. This air
ACTIVATES the valves; it doesn't pass through the valve.

Cheers,
Henk

On 2/23/2014 9:59 AM, microscopylistserver-noreply-at-microscopy.com wrote:
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} Email: jcsmtf-at-mail.missouri.edu
} Name: Josh Schorp
}
} Organization: University of Missouri
}
} Title-Subject: [Filtered] CM10 pneumatics
}
} Message: I am receiving a Pneumatics message on the screen of our CM10 and I believe this to be the
} reason while none of the vacuums will start. The air line on our machine runs first to V11, but it
} has not been allowed past V11. Is there a reason for this? I'm not sure how this valve works, I took
} it off and it seems to be just a metal unit with no moving parts. I'm guessing the black electrical
} piece that connects to it has something to do with its operations. Does anyone have any insight on
} how to open this valve? I am running ~90 psi to it.
}
} Login Host: 161.130.188.19
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Hi Josh,

The microscope air supply should *NOT* be attached to V11. V11 is the
microscope vent valve and is used to release the vacuum in the column
and camera chamber. It should never have more than a fraction of a psi
supplied to it. Higher pressure can force the viewing chamber window
out with great force and cause serious injury to anyone nearby! Venting
the column with high pressure air when the sample rod is inserted can
also blow the sample rod out with some force and damage it.

The air for the valves is supplied through the "Watts" regulator to the
distribution block on the rear of the microscope column. This air
ACTIVATES the valves; it doesn't pass through the valve.

Cheers,
Henk

On 2/23/2014 9:59 AM, microscopylistserver-noreply-at-microscopy.com wrote:
}
}
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
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} please copy both jcsmtf-at-mail.missouri.edu as well as the Microscopy Listserver
} ---------------------------------------------------------------------------
}
} Email: jcsmtf-at-mail.missouri.edu
} Name: Josh Schorp
}
} Organization: University of Missouri
}
} Title-Subject: [Filtered] CM10 pneumatics
}
} Message: I am receiving a Pneumatics message on the screen of our CM10 and I believe this to be the
} reason while none of the vacuums will start. The air line on our machine runs first to V11, but it
} has not been allowed past V11. Is there a reason for this? I'm not sure how this valve works, I took
} it off and it seems to be just a metal unit with no moving parts. I'm guessing the black electrical
} piece that connects to it has something to do with its operations. Does anyone have any insight on
} how to open this valve? I am running ~90 psi to it.
}
} Login Host: 161.130.188.19
} Listserver Email Form V - 20120416
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==============================Original Headers==============================
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Dear members of the listserver:

We have a very old SEM XL20 (FEI, runs under Windows 3.11, engl. version). Our hard disk crashed and we don't have a backup.

We have replaced the hard disk and got MS DOS and Win 3.11. installed, but we have problems with the operating system for the SEM. FEI is not able to help us, because the XL20 is so old.

Thus we are looking for:
- a backup of the XL20 operating system or
- XL20 - software V 5.24 (on discs or CD-ROM) including driver software or
- EZ-SCSI-Driver (v3). (ADAPTEC-SCSI-SOFTWARE) or
- MACH 64- Driver or
- ATI-Driver for Win 3.11 (engl. Version).

Sincerely, Elisabeth Liebler-Tenorio

==============================Original Headers==============================
9, 29 -- From Elisabeth.Liebler-Tenorio-at-fli.bund.de Wed Feb 26 04:57:35 2014
9, 29 -- Received: from mx022.fli.bund.de (mx022.fli.bund.de [193.22.112.22])
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9, 29 -- mapi id 14.03.0174.001; Wed, 26 Feb 2014 11:57:33 +0100
9, 29 -- From: "Liebler-Tenorio, Elisabeth" {Elisabeth.Liebler-Tenorio-at-fli.bund.de}
9, 29 -- To: "Microscopy-at-Microscopy.Com" {Microscopy-at-Microscopy.Com}
9, 29 -- CC: "ethalmann-at-t-online.de" {ethalmann-at-t-online.de}
9, 29 -- Subject: SEM XL20 (FEI) operating system/drivers
9, 29 -- Thread-Topic: SEM XL20 (FEI) operating system/drivers
9, 29 -- Thread-Index: Ac8y4WJQgviqvi0tQz6N3f8e25e20g==
9, 29 -- Date: Wed, 26 Feb 2014 10:57:32 +0000
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Dear Elisabeth,
I found some old diskettes with following ATI MACH-64 drivers for
Win3.11:

ATI Mach-64_2012
ATI Mach-64_222
ATI Mach-64_303

If you like, I can send you an archiv (zip) with all the files I have
found on them.

Best regards from Prague

Oldrich

--
Oldrich Benada
Institute of Microbiology AS CR, v.v.i.
Videnska 1083
142 20 Prague 4
Czech Republic

On Wed, 26 Feb 2014 05:02:18 -0600,
Elisabeth.Liebler-Tenorio-at-fli.bund.de wrote :
}
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} ----------------------------------------------------------------------------
}
} Dear members of the listserver:
}
} We have a very old SEM XL20 (FEI, runs under Windows 3.11, engl.
} version). Our hard disk crashed and we don't have a backup.
}
} We have replaced the hard disk and got MS DOS and Win 3.11.
} installed, but we have problems with the operating system for the
} SEM. FEI is not able to help us, because the XL20 is so old.
}
} Thus we are looking for:
} - a backup of the XL20 operating system or
} -Â XL20 - software V 5.24 (on discs or CD-ROM) including driver
} software or
} -Â EZ-SCSI-Driver (v3). (ADAPTEC-SCSI-SOFTWARE) or
} -Â MACH 64- Driver or
} -Â ATI-Driver for Win 3.11 (engl. Version).
}
} Sincerely, Elisabeth Liebler-Tenorio
}
}
}
}
}
} ==============================Original
} Headers============================== 9, 29 -- From
} Elisabeth.Liebler-Tenorio-at-fli.bund.de Wed Feb 26 04:57:35 2014 9, 29
} -- Received: from mx022.fli.bund.de (mx022.fli.bund.de
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} +0100 9, 29 -- From: "Liebler-Tenorio, Elisabeth"
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} "Microscopy-at-Microscopy.Com" {Microscopy-at-Microscopy.Com} 9, 29 -- CC:
} "ethalmann-at-t-online.de" {ethalmann-at-t-online.de} 9, 29 -- Subject: SEM
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} (FEI) operating system/drivers 9, 29 -- Thread-Index:
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==============================Original Headers==============================
8, 40 -- From benada-at-biomed.cas.cz Wed Feb 26 08:09:01 2014
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8, 40 -- Date: Wed, 26 Feb 2014 15:08:56 +0100
8, 40 -- From: Oldrich Benada {benada-at-biomed.cas.cz}
8, 40 -- To: Elisabeth.Liebler-Tenorio-at-fli.bund.de, Microscopy-at-Microscopy.com
8, 40 -- Subject: Re: [Microscopy] SEM XL20 (FEI) operating system/drivers
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Dear Elisabeth,

Last summer I had fun recovering WIN-NT PC on FEI FIB620 (Phillips XL30
SEM with ion beam column added) from a hard disk crash.

If your original disk is physically operational, then recovering SEM
software from it may still be possible. Not sure if PC "hackery" is of
interest for everyone, so please contact me off list you want/need to
give it a try.

Valery Ray
=================================
PBS&T, MEO Engineering Co., Inc.
290 Broadway, Suite 298
Methuen, MA 01844 USA
Phone: +1-978-296-5063 - leave a message with call-back number
US Mobile: +1-978-305-0479
Skype: pbstmeo
E-mail: vray-at-partbeamsystech.com
Web: www.partbeamsystech.com
Web: www.freudlabs.com

On 2/26/2014 5:59 AM, Elisabeth.Liebler-Tenorio-at-fli.bund.de wrote:
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
} To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
} On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} ----------------------------------------------------------------------------
}
} Dear members of the listserver:
}
} We have a very old SEM XL20 (FEI, runs under Windows 3.11, engl. version). Our hard disk crashed and we don't have a backup.
}
} We have replaced the hard disk and got MS DOS and Win 3.11. installed, but we have problems with the operating system for the SEM. FEI is not able to help us, because the XL20 is so old.
}
} Thus we are looking for:
} - a backup of the XL20 operating system or
} - XL20 - software V 5.24 (on discs or CD-ROM) including driver software or
} - EZ-SCSI-Driver (v3). (ADAPTEC-SCSI-SOFTWARE) or
} - MACH 64- Driver or
} - ATI-Driver for Win 3.11 (engl. Version).
}
} Sincerely, Elisabeth Liebler-Tenorio
}
}
}
}
}
} ==============================Original Headers==============================
} 9, 29 -- From Elisabeth.Liebler-Tenorio-at-fli.bund.de Wed Feb 26 04:57:35 2014
} 9, 29 -- Received: from mx022.fli.bund.de (mx022.fli.bund.de [193.22.112.22])
} 9, 29 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id s1QAvYMK027973
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} 9, 29 -- Received: from FLI-JE-MB031.intern.fli.bund.local (172.18.0.31) by
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} 9, 29 -- mapi id 14.03.0174.001; Wed, 26 Feb 2014 11:57:33 +0100
} 9, 29 -- From: "Liebler-Tenorio, Elisabeth" {Elisabeth.Liebler-Tenorio-at-fli.bund.de}
} 9, 29 -- To: "Microscopy-at-Microscopy.Com" {Microscopy-at-Microscopy.Com}
} 9, 29 -- CC: "ethalmann-at-t-online.de" {ethalmann-at-t-online.de}
} 9, 29 -- Subject: SEM XL20 (FEI) operating system/drivers
} 9, 29 -- Thread-Topic: SEM XL20 (FEI) operating system/drivers
} 9, 29 -- Thread-Index: Ac8y4WJQgviqvi0tQz6N3f8e25e20g==
} 9, 29 -- Date: Wed, 26 Feb 2014 10:57:32 +0000
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5, 43 -- From vray-at-partbeamsystech.com Wed Feb 26 08:23:26 2014
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Hello all,

A user of our JEOL 2100 TEM informed me that he was noticing anomalous
rings appearing in diffraction patterns acquired with the SAD aperture
centered over vacuum (he's developing a quantitative diffraction
technique and presumably is acquiring a background diffraction
spectrum). I triple checked the microscope alignment in TEM mode,
STEM, nanobeam diffraction, convergent beam diffraction, and the high
current EDS mode are all well aligned over the full range of spot
sizes and convergence angles. The lens voltages and alignment coils
all fall within their suitable ranges as well. When I acquired
diffraction patterns of actual sample I don't see the artifacts in the
DP, presumably because the intensity of the actual diffraction
rings/spots washes out the weak artifact rings.

I checked all the condensor and selected area apertures, and all
exhibit the problem as far as I can tell. I also triple checked to
ensure all other apertures were removed and not displaced into a wrong
position by a user, and they all checked out as well.

I'm attaching a link to two images of the artifact. These were taken
at 200kV but the same is seen at 120kV (and probably 80kV--have not
checked). You can see that there are two rings and their size and
spacing change with changing camera lengths. Is this caused by
diffraction of something in the beam path--either an aperture carrier
or lost sample--or another effect, or am I missing something simple?
Thanks for any hints or help directed my way.

Link to DPs showing artifacts: http://imgur.com/a/xCV4R

Thanks,
Chris
Virginia Tech

==============================Original Headers==============================
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6, 26 -- Subject: Anomalous diffraction rings in JEOL 2100
6, 26 -- From: Christopher Winkler {microwink-at-gmail.com}
6, 26 -- To: Microscopy-at-microscopy.com
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On Feb 26, 2014, at 3:34 PM, microwink-at-gmail.com wrote:

} A user of our JEOL 2100 TEM informed me that he was noticing anomalous
} rings appearing in diffraction patterns acquired with the SAD aperture
} centered over vacuum (he's developing a quantitative diffraction
} technique and presumably is acquiring a background diffraction
} spectrum). I triple checked the microscope alignment in TEM mode,
} STEM, nanobeam diffraction, convergent beam diffraction, and the high
} current EDS mode are all well aligned over the full range of spot
} sizes and convergence angles. The lens voltages and alignment coils
} all fall within their suitable ranges as well. When I acquired
} diffraction patterns of actual sample I don't see the artifacts in the
} DP, presumably because the intensity of the actual diffraction
} rings/spots washes out the weak artifact rings.
}
} I checked all the condensor and selected area apertures, and all
} exhibit the problem as far as I can tell. I also triple checked to
} ensure all other apertures were removed and not displaced into a wrong
} position by a user, and they all checked out as well.
}
} I'm attaching a link to two images of the artifact. These were taken
} at 200kV but the same is seen at 120kV (and probably 80kV--have not
} checked). You can see that there are two rings and their size and
} spacing change with changing camera lengths. Is this caused by
} diffraction of something in the beam path--either an aperture carrier
} or lost sample--or another effect, or am I missing something simple?
} Thanks for any hints or help directed my way.
}
} Link to DPs showing artifacts: http://imgur.com/a/xCV4R
}
} Thanks,
} Chris
} Virginia Tech
}
Dear Chris,
I noticed that the rings are not centered with the undiffracted
beam. Also, the pattern appears to be from a polycrystalline
specimen. It may be that the beam emitted by the FEG/filament is
split in two--either by having two points emitting from the source or
by the beam hitting something near the top of the column producing a
scattered beam. If this second beam strikes some polycrystalline
substance, such as a grid near the point of vacuum where the main beam
is directed, it could produce something like the DPs on the web site.
Do the DPs change when the stage is moved (while keeping the beam over
vacuum, of course)?
Yours,
Bill

==============================Original Headers==============================
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6, 37 -- Subject: Re: [Microscopy] Anomalous diffraction rings in JEOL 2100
6, 37 -- Date: Wed, 26 Feb 2014 18:49:03 -0800
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Yes, I used to see these on our JEOL 2500 when I was at HP. Tomba San, Mr Tomba at JEOL fixed it by always having us do diffraction left of crossover- where the beam is more parallel! That will most likely give the heave ho to your rings!

Ganbette
Good Luck or Hang in there!

Pete Eschbach
Oregon State EM Facility

Sent from my iPhone

} On Feb 26, 2014, at 3:35 PM, microwink-at-gmail.com wrote:
}
}
}
}
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} Hello all,
}
} A user of our JEOL 2100 TEM informed me that he was noticing anomalous
} rings appearing in diffraction patterns acquired with the SAD aperture
} centered over vacuum (he's developing a quantitative diffraction
} technique and presumably is acquiring a background diffraction
} spectrum). I triple checked the microscope alignment in TEM mode,
} STEM, nanobeam diffraction, convergent beam diffraction, and the high
} current EDS mode are all well aligned over the full range of spot
} sizes and convergence angles. The lens voltages and alignment coils
} all fall within their suitable ranges as well. When I acquired
} diffraction patterns of actual sample I don't see the artifacts in the
} DP, presumably because the intensity of the actual diffraction
} rings/spots washes out the weak artifact rings.
}
} I checked all the condensor and selected area apertures, and all
} exhibit the problem as far as I can tell. I also triple checked to
} ensure all other apertures were removed and not displaced into a wrong
} position by a user, and they all checked out as well.
}
} I'm attaching a link to two images of the artifact. These were taken
} at 200kV but the same is seen at 120kV (and probably 80kV--have not
} checked). You can see that there are two rings and their size and
} spacing change with changing camera lengths. Is this caused by
} diffraction of something in the beam path--either an aperture carrier
} or lost sample--or another effect, or am I missing something simple?
} Thanks for any hints or help directed my way.
}
} Link to DPs showing artifacts: http://imgur.com/a/xCV4R
}
} Thanks,
} Chris
} Virginia Tech
}
} ==============================Original Headers==============================
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} 6, 26 -- Subject: Anomalous diffraction rings in JEOL 2100
} 6, 26 -- From: Christopher Winkler {microwink-at-gmail.com}
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} 6, 26 -- Content-Type: text/plain; charset=ISO-8859-1
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7, 35 -- Subject: Re: [Microscopy] Anomalous diffraction rings in JEOL 2100
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The deadline for abstract submission has passed and as you are making
plans to attend the Hartford meetings Aug. 3-7, please don*t forget
about MSA*s student bursary program.
Its purpose is to encourage students to attend the meetings by helping
to defray some of the costs; giving them an opportunity to meet and
interact with the established microscopy community.

The students will be paid $10 an hour to work for ~20 hours (up to 40
hours) during the meeting or pre-meeting events. The jobs involve such
things as providing support in the different symposia (assisting with
audio-visual needs, speaker set-up, maintaining an attendance count),
staffing the volunteer office, monitoring use of the Internet Café, and
helping with vendor tutorial sign-up. Payment is given as a check at
the end of the meetings or when the student leaves Hartford.

Once the program has been finalized, each registered bursary will be
contacted and allowed to choose the times and activities they would like
to work. Many times they end up *working* sessions they would
attend anyway. There is an added bonus of $10 cash to help with meals
for each morning and/or afternoon session worked and a meeting shirt.

If anyone would like to participate in the bursary program, please
check the *I wish to apply for a student bursary* box in section 2
of the registration form and send me an email of your intention.
Bursary space is limited, so sign-up early. Applicants for the
bursaries must be members of MSA or MAS, enrolled as students at a
recognized educational institution, and have paid their registration
fee.

For those *non-students* volunteers are also needed to help with
the above mentioned meeting activities. Although not paid on an hourly
basis as the student bursaries, volunteers do receive a meeting shirt
and the same cash allotment to help with meals. Volunteers also have
the opportunity to interact more with the microscopy community as they
assist with meeting tasks.

If anyone has any questions about the bursary/volunteer program, or
would like to participate, please contact:

Amanda Lawrence
Institute for Imaging and Analytical Technologies
Mississippi State University
662-325-7998
alawrence-at-i2at.msstate.edu

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Email: lamiller-at-illinois.edu
Name: Lou Ann Miller

Organization: Materials Research Lab, U of Illinois

Title-Subject: [Filtered] MRL Biological microscopy and analysis work shop Opportunity for vendors,
and for students

Message: Hello!

Many thanks to many of you for helping out with our fall biological conference in 2013!

This year, we hope to have the conference November 12 & 13th, 2014.

Students: There will be a poster and presentation Competition at this year's MRL Biological
microscopy and analysis work shop.

Each year we have had a different theme, 1st Biostructures and "what are we looking at", last
year Techniques: " how do we do that?"

This year, I'm hoping to pull in more students and participation by having a student presentation
and poster competition as well as talks by those who have specialties here on campus.. The subject
will be how students put biological microscopy techniques into use.

VENDORS:

We would like to offer prizes for the best presentations with posters. Of course this is where you
can come in. :-)

Because you are vendors with the university, we checked all the rules, and it appears that a
check from a vendor straight to the prize winner can be up to , but not exceed $100.

The first 3 vendors, who can offer a nice monetary prize($100 or less), will get to be on the
judges panel, as well as have the prize named after your company. We can send the results with
images with your company's name to our paper, and to the university paper.

Vendors, please let me know by March 30th if you are interested.

If you miss this, and still would like mention around the conference, feel free to email me about
sponsoring a break for food.

Students, more information will be forthcoming for you. With biological applications also mixing
with material science these days, you may be interested if you mix the two disciplines as well.

Thanks!

Lou Ann Miller
Biology Specialist, Frederick Seitz Materials Research Lab

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Email: ghoshroy-at-sc.edu
Name: Soumitra Ghoshroy

Organization: University of South Carolina

Title-Subject: [Filtered] Chromium sputtering target

Message: Dear Colleagues,

We have Denton DESK II Turbo Sputter Coater and are planning to use a chromium sputtering target for
the first time. We intend to use this coating for FESEM observation of a variety of specimens.We are
familiar with gold and gold-palladium targets but never used a chromium target before. This morning
we coated a sample with chromium, but seems like it didn't get any coating at all. I did see the
plasma. When we use gold or gold-palladium in the same coater, it works fine and we can see the
coating on the surface. Is it possible to see a color difference after Cr coating for 60-100 sec. We
are using 40-60% power.I noticed the Cr target is much thicker than the gold target.

I will appreciate someone telling if we are doing something wrong or it is just our simple ignorance
about chromium target. Any help will be very much appreciated.

Thanks in advance,

Soumitra

Soumitra Ghoshroy
EM Center
University of South Carolina
Columbia, SC 29208

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Email: gcat84-at-gmail.com
Name: Glenn

Organization: University of Delaware

Title-Subject: [Filtered] ISI DS-130 - schematics

Message: Hello,

I have an ISI DS-130 in mostly working condition. However, it recently gave up on producing some of
the +/- 15 V outputs. The first indication was when the vacuum would kick out when enabling the
filament. Then, the emission displays 200 uA even when the voltage and current are set to zero.

Does anyone have original wiring diagrams for this model SEM unit? It would be greatly appreciated.

Regards,
Glenn

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Hi

I do not know the Denton Turbo Sputter Coater, but I was involved with the
development of Chromium coating many years ago!

To sputter chromium is not like sputtering any of the more familiar
materials. Firstly, you need a high vacuum and a power supply that is rated
much higher than for the conventional sputtering materials. Secondly,
before you are able to sputter the chromium the oxide coating has to be
removed, and this has to happen every time you use the unit. So the first
part of the sputter process is oxide removal, most dedicated systems have a
shutter to catch the oxide, thus leave the specimens clean. You know when
all of the oxide has been removed as the plasma will have a nice pale blue
colour. Once you have this level of target cleaning you switch off the
plasma, remove the protective shutter, and coat for about 2 to 5 seconds.
You should not expect to be able to see the coating.

It's a very long time since I was involved with this work but I hope I have
been able to provide some assistance?

Good luck

Steve

Steve Chapman FRMS
Protrain for Consultancy and Courses in Electron Microscopy
Mobile +44 (0) 7711 606967 Fax +44 (0)1280 814007
www.emcourses.com

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

Email: ghoshroy-at-sc.edu
Name: Soumitra Ghoshroy

Organization: University of South Carolina

Title-Subject: [Filtered] Chromium sputtering target

Message: Dear Colleagues,

We have Denton DESK II Turbo Sputter Coater and are planning to use a
chromium sputtering target for the first time. We intend to use this coating
for FESEM observation of a variety of specimens.We are familiar with gold
and gold-palladium targets but never used a chromium target before. This
morning we coated a sample with chromium, but seems like it didn't get any
coating at all. I did see the plasma. When we use gold or gold-palladium in
the same coater, it works fine and we can see the coating on the surface. Is
it possible to see a color difference after Cr coating for 60-100 sec. We
are using 40-60% power.I noticed the Cr target is much thicker than the gold
target.

I will appreciate someone telling if we are doing something wrong or it is
just our simple ignorance about chromium target. Any help will be very much
appreciated.

Thanks in advance,

Soumitra

Soumitra Ghoshroy
EM Center
University of South Carolina
Columbia, SC 29208

==============================Original Headers==============================
20, 24 -- From protrain-at-emcourses.com Fri Feb 28 08:03:44 2014
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Dear all,

We are working with an ESEM Quanta 600 in the low vacuum mode. We need a pressure of 0.18 Torr, but when we choose it via the UI the pressure is not constant but jumps between ~0.17 and ~0.28 Torr.
At our second ESEM Quanta 200 we experience no problems with the pressure stability at 0.18 Torr.

Do you have an idea, which parameter or defect is responsible?

Thank you in advance!

Stefan

==============================Original Headers==============================
6, 30 -- From stefan.wernitznig-at-medunigraz.at Fri Feb 28 08:36:34 2014
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Soumitra, as a compromise might I suggest using Pt, it will definitely give you better results than Au or Au/Pd.

Good Luck,
John Robson

-----Original Message-----
X-from: protrain-at-emcourses.com [mailto:protrain-at-emcourses.com]
Sent: Friday, February 28, 2014 9:13 AM
To: Robson,John (AN) BIP-US-R

Hi

I do not know the Denton Turbo Sputter Coater, but I was involved with the development of Chromium coating many years ago!

To sputter chromium is not like sputtering any of the more familiar materials. Firstly, you need a high vacuum and a power supply that is rated much higher than for the conventional sputtering materials. Secondly, before you are able to sputter the chromium the oxide coating has to be removed, and this has to happen every time you use the unit. So the first part of the sputter process is oxide removal, most dedicated systems have a shutter to catch the oxide, thus leave the specimens clean. You know when all of the oxide has been removed as the plasma will have a nice pale blue colour. Once you have this level of target cleaning you switch off the plasma, remove the protective shutter, and coat for about 2 to 5 seconds.
You should not expect to be able to see the coating.

It's a very long time since I was involved with this work but I hope I have been able to provide some assistance?

Good luck

Steve

Steve Chapman FRMS
Protrain for Consultancy and Courses in Electron Microscopy
Mobile +44 (0) 7711 606967 Fax +44 (0)1280 814007
www.emcourses.com

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

Email: ghoshroy-at-sc.edu
Name: Soumitra Ghoshroy

Organization: University of South Carolina

Title-Subject: [Filtered] Chromium sputtering target

Message: Dear Colleagues,

We have Denton DESK II Turbo Sputter Coater and are planning to use a chromium sputtering target for the first time. We intend to use this coating for FESEM observation of a variety of specimens.We are familiar with gold and gold-palladium targets but never used a chromium target before. This morning we coated a sample with chromium, but seems like it didn't get any coating at all. I did see the plasma. When we use gold or gold-palladium in the same coater, it works fine and we can see the coating on the surface. Is it possible to see a color difference after Cr coating for 60-100 sec. We are using 40-60% power.I noticed the Cr target is much thicker than the gold target.

I will appreciate someone telling if we are doing something wrong or it is just our simple ignorance about chromium target. Any help will be very much appreciated.

Thanks in advance,

Soumitra

Soumitra Ghoshroy
EM Center
University of South Carolina
Columbia, SC 29208

==============================Original Headers==============================
20, 24 -- From protrain-at-emcourses.com Fri Feb 28 08:03:44 2014 20, 24 -- Received: from mail.esentra.net (mail.esentra.net [94.228.36.208])
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Soumitra;
Chromium is extremely reactive with oxygen. Even with a UHV system, chromium deposits tend to be chromium oxide. The best way to deposit chromium is with an ion beam sputtering system. Plasma sputtering, as you have discovered, does not work well for chromium. You see the glow of the plasma, but not much else happens.

John Mardinly, ASU

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Email: ghoshroy-at-sc.edu
Name: Soumitra Ghoshroy

Organization: University of South Carolina

Title-Subject: [Filtered] Chromium sputtering target

Message: Dear Colleagues,

We have Denton DESK II Turbo Sputter Coater and are planning to use a chromium sputtering target for the first time. We intend to use this coating for FESEM observation of a variety of specimens.We are familiar with gold and gold-palladium targets but never used a chromium target before. This morning we coated a sample with chromium, but seems like it didn't get any coating at all. I did see the plasma. When we use gold or gold-palladium in the same coater, it works fine and we can see the coating on the surface. Is it possible to see a color difference after Cr coating for 60-100 sec. We are using 40-60% power.I noticed the Cr target is much thicker than the gold target.

I will appreciate someone telling if we are doing something wrong or it is just our simple ignorance about chromium target. Any help will be very much appreciated.

Thanks in advance,

Soumitra

Soumitra Ghoshroy
EM Center
University of South Carolina
Columbia, SC 29208

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Dear Colleagues,

On behalf of the organizing committee, Iâ€™d like to extend an
invitation for you to attend the first Microscopy of Infectious
Disease Agents Symposia we are hosting at Rocky Mountain
Laboratories/NIAID/NIH in Hamilton, Montana July 24th and 25th.
Please feel free to pass this along to anyone who might be interested
in attending.

The 2014 Microscopy of Infectious Disease Agents Symposia (MIDAS) will
provide an opportunity to bring together leading researchers of
microscopic imaging and related techniques as they apply to the study
of human pathogens. Organized by the National Institute of Allergy of
Infectious Diseases, National Institutes of Health, this meeting will
provide a venue for recent progress in areas of both light and
electron microscopies relevant to imaging viruses, bacteria, prions,
host cells and tissues. In addition to scientific presentations,
workshops will be organized to teach various aspects of emerging
technologies. The field of microscopic imaging is rapidly evolving
as advances in both instrumentation and computational sciences are
enabling a greatly expanding range of analyses to be performed. A
broad theme at the conference will be on technological advances and on
future developments needed to increase the application and impact of
microscopic imaging for understanding the biology of human pathogens
with the goal of improving the treatment, diagnosis, and prevention of
infectious diseases. The conference will bring together a diverse
group of scientists at senior and junior levels. Seminars and posters
at the conference will provide an opportunity to present research
findings and exchange of ideas.

Please visit our registration site for more information and the
preliminary agenda.
respond.niaid.nih.gov/conferences/rmlsymposium2014/Pages

Keynote speakers:

Sriram Subramaniam, Ph.D., National Cancer Institute, NIH
â€¨Dr. Sriram Subramaniam received his Ph.D. in Physical Chemistry from
Stanford University and completed postdoctoral training in the
Departments of Chemistry and Biology at M.I.T. He is chief of the
Biophysics Section in the Laboratory of Cell Biology at the Center for
Cancer Research, National Cancer Institute. He holds a visiting
faculty appointment at the Johns Hopkins University School of
Medicine. His current work is focused on the development of advanced
technologies for imaging macromolecular assemblies using 3D electron
microscopy, and their application to address fundamental problems in
HIV/AIDS and cancer research.
Hari Shroff , Ph.D., National Ins titute of Biomedical Imaging and
Bioengineering, NIH
Dr. Hari Shroff received a B.S.E. in bioengineering from the
University of Washington in 2001, and under the supervision of Dr. Jan
Liphardt, completed his Ph.D. in biophysics at the University of
California at Berkeley in 2006. He spent the next three years
performing postdoctoral research under the mentorship of Eric Betzig
at the Howard Hughes Medical Institute's Janelia Farm Research Campus
where his research focused on development of photo-activated
localization microscopy (PALM), an optical super resolution technique.
Dr. Shroff is now chief of NIBIB's section on high resolution optical
imaging laboratory, where he and his staff are developing new imaging
tools for application in biological and clinical research. Dr
Shroffâ€™s lab has recently developed two new microscope systems: The
first, iSIM, uses structured illumination and high speed processing to
render images with double the resolution of conventional fluorescence
microscope. The second, diSPIM, is a light sheet microscope which
uses two objectives to achieve isotropic resolution with minimal
bleaching.

Organizing committee:
Elizabeth Fischer, NIH/NIAID, Audray Harris, NIH/NIAID, and Owen
Schwartz, NIH/NIAID

Contact:
Elizabeth R. Fischer
Microscopy Unit Manager
Research Technologies Section/RTB
Rocky Mountain Laboratories/NIAID/NIH
903 South 4th Street
Hamilton, MT 59840
406-363-9378

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Email: tina.williams-at-ars.usda.gov
Name: Tina Williams

Organization: USDA

Title-Subject: [Filtered] vacuum oven polymerization: Vent to hood?

Message: Hello,

While polymerizing your resins, do you vent your oven in a fume hood or to the lab? Safety
suggestions?
Thank you,
Tina

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Dear Listers,

PITTCON is not typically considered an important venue for microscopy, but
there will be at least three dozen microscopy and related technology
companies exhibiting at Chicago's McCormick Center, March 2-6. In
addition, there will be a number of seminars, tutorials, and lunch&learn
opportunities. American Lab asked that I do a review of Microscopy at
PITTCON. That article is now posted at:

http://www.americanlaboratory.com/913-Technical-Articles/156565-Integrating-Microscopy-Into-the-Analytical-Scheme-A-Pittcon-2014-Microscopy-Review/

Caveat: I have no commercial interest in any of the companies represented
here.

Best regards,
Barbara Foster
Microscopy/Microscopy Education
P: (972)924-5310 E: bfoster-at-mme1.com

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realname - Pamela Coker, Ph.D.
Email - pjcoker-at-pima.edu
ORGANIZATION - Ph.D.
EDUCATION - Undergraduate College
LOCATION - Tucson, AZ 85743
SUBJECT_OF_QUESTION - Looking for a reference slide set for pharmaceuticals
QUESTION - Hello,

I am writing a proposal for a new course in forensic microscopy. I have
found most of the reference slide sets I need, however I am coming up
empty handed on a pharmaceutical slide set.

Can anyone point me in the right direction? I'm not finding one at
McCrone's.
Thanks, Dr. C.

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Hitachi owners,

I was wondering if there are digital versions of the user manuals for the
Hitachi 7000 transmission EM and Hitachi S-3000N scanning EM? If so, I
would very much appreciate a copy of them. Please contact me off-line and
we can see how this can be arranged.

Many thanks,
Debby

Debra Sherman, Founder & CSO
DS imaging LLC
Purdue Kurz Technology Center
1281 Win Hentschel Blvd
West Lafayette, IN 47906
E-mail: debby.sherman-at-dsimagingllc.com
www.dsimagingllc.com
Mobile: 765-418-8540

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Group,

Trying to address an electronics high temperature error on our Leo 906E TEM.

Would like to know where the temperature sensor is located on the system (column chassis or power supply rack) in order to troubleshoot the problem further.

Have thought for some time the issue was with the chiller but its running at 58 degrees F at a flow rate of 54 gph and the room is 70 degrees F.

On our coldest days this winter we have successfully run the instrument but we had a warm spell last week and the problem reappeared.

All rack cooling fans are running and filters are clean.

Any ideas are appreciated.

Roy Beavers
Southern Methodist University
rbeavers-at-smu.edu

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Email: tina.williams-at-ars.usda.gov
Name: Tina Williams

Organization: USDA

Title-Subject: [Filtered] vacuum oven polymerization: Vent to hood- Thanks for all the
replies/suggestions

Message: Hello again,

Thank you to all who replied to my question regarding venting in a fume hood during polymerization
of resins. I especially appreciated the references and safety reminders to reduce
respiratory/skin/immuno sensitivities by venting in a fume hood.

I wanted to confirm, by having your collective experiences working with resins, that we continue
working in harmony with these safety protocols. We have been venting the resin polymerization in
the fume hood. However our current EHS officer, brought up the concern regarding the oven
effecting the airflow in the hood. Since we have a large/heavy oven, and the same issue may apply
with a small oven (if one could be found), we have kept the oven in the hood.

Your answers help us have a basis to continue with our current safety protocols, until a safer
alternative is found.

Tina

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Email: gr3g0rr33-at-gmail.com
Name: Gregory Wyche

Organization: University of South Carolina

Title-Subject: [Filtered] Networking Opportunities In CT

Message: Hi, I'm a microscopist in CT (currently between jobs) looking for opportunities to network
in that state. The Connecticut Microscopy Society website has not been updated in a long time so I
can't get in touch with any of the members. If you know of any groups that meet, please forward
them. I'll very much appreciate it.

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Email: dvanhart-at-binghamton.edu
Name: Daniel VanHart

Organization: Binghamton University

Title-Subject: [Filtered] FTIR Microscope

Message: Does anyone have any experience with a Pike Technologies microMax Microscope?

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Email: gul417-at-mail.usask.ca
Name: Guosheng Liu

Organization: University of Saskatchewan

Title-Subject: [Filtered] TEM (Philips CM10) service contract provider?

Message: Hello EM Community,

We are shopping for a service contractor for our CM10 TEM, ideally from Canada or USA. This
20-year-old but sill reliable TEM has been service-contracted for most of its years but run without
contract since early 2013(budge issue).....I was wondering if you would like to share information
regarding service engineers/companies for the same or similar equipment. We will purchase a service
coverage either very basic or all-inclusive depending on the quotes.

Thanks a lot.

Guosheng Liu

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Greg

You do know that the largest and most extensive microscopy meeting in the
world will be held in Hartford CT, this August right?

http://www.microscopy.org/MandM/2014/index.cfm

There will certainly be opportunities for you to make connections there.

Nestor
Your Friendly Neighborhood SysOp

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} in that state. The Connecticut Microscopy Society website has not been updated in a long time so I
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===========================================
Dr. Nestor J. Zaluzec
Argonne National Laboratory
Electron Microscopy Center
Materials Science/Bldg 212
9700 S. Cass Ave.
Argonne, Illinois 60439 USA
Tel: 530-NES-TORZ (530-637-8679) Fax: 630-252-4798

iChat:Zaluzec-at-AIM
Skype: Zaluzec-at-ANL
TPM: http://tpm.amc.anl.gov

Email: Zaluzec-at-aaem.amc.anl.gov
WWW: http://tpm.amc.anl.gov

Senior Scientist - Argonne National Laboratory
Fellow of the Microscopy Society of America
Senior Fellow of the Computational Institute - University of Chicago
E.P. Wigner Fellow - Oak Ridge National Laboratory
Past-President: Microscopy Society of America
Adjunct Professor of Physics - Northern Illinois University
and University of Illinois at Chicago
Visiting Professor of Materials Characterziation, Manchester University UK.

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

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Gregory -

Since you've used the web inquiry box to submit your question, I
assume that you're not a MSA member. That's a big mistake. If you
were a member, you could access a membership list that would give you
contact information on 39 Connecticut members. Plus a very efficient
job placement service. If you join now, you can also register for
the Hartford meeting at the members' rate, a substantial saving.

Caroline

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--
Caroline Schooley
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45301 Caspar Point Road, Box 117
Caspar, CA 95420
Phone/FAX (707)964-9460
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If you use Google to search for Airy disk, and select the reference
to the Wikipedia entry you will find a very extensive coverage of
the subject of the Airy disk

While I'm at it, I can't resist including a story relating to this
topic that I used to tell in my class when discussing TEM resolution;

A Cockney lady from London was touring New York City, and was taking
a ride on the NY Subway. Now the subway trains push a rather strong
blast of air ahead of them as they travel through the subway tunnels.
When they approach a station this blast of air flows up through the
opening creating quite a strong breeze. Well, our lady was alighting
from one subway train just as another pulled into the station. The
blast of air generated by this approaching train caught her skirts
and blew them sky high, revealing the fact that she was not wearing
any underwear. She, of course, acted terribly embarrassed, and a
conductor standing nearby and seeing her distress thought he should
say something to smooth over the situation. So, he said, "Airy,
isn't it?" to which the lady replied in her native Cockney
accent,"Wot in 'ell did ye expect, feathers?"
--
Wilbur C. Bigelow, Professor Emeritus
Materials Sci. & Engr., Univ. of Michigan
Ann Arbor, Michigan 48109-2136
e-mail: bigelow-at-umich.edu;
Fx:734-763-4788; Ph:734-975-0858
Address mail to: 2911 Whittier Court
Ann Arbor, MI 48104-6731

==============================Original Headers==============================
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Contents Retrieved from Microscopy Listserver Archives
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Dear all,

We are pleased to announce

the 4th UniBe Practical Course on Cryo-Electron Microscopy of Vitreous
Sections (CEMOVIS, a.k.a cryoEM of frozen-hydrated sections)

11-13 August 2014 / 14-16 August 2014

Organiser : Benoît Zuber
Instructors: Benoît Zuber, Daniel Studer, Ioan Iacovache

The objective of the course is to teach the participants the practical
skills necessary to successfully apply CEMOVIS in their laboratories.
Essential background theory of CEMOVIS will be given and most of the
time will be spent practicing high-pressure freezing, cryo-sectioning,
and low-dose TEM imaging. 1 high-pressure freezing machine, 3
state-of-the-art cryo-ultramicrotomes and 1 cryo-electron microscope
will be dedicated to the course. Of special interest, participants
will learn to use the new tools that we developed to facilitate
cryosectioning (Studer et al 2014 J Struct Biol 185(1):125-8). The
3-day course is given twice, each time with a maximum of 4
participants so that they can be actively practicing at any time.

The course is intended for scientists whose research projects will
benefit from the use of CEMOVIS. Experience in either cryo-electron
microscopy or ultramicrotomy of resin-embedded specimens is a
prerequisite.

Information and registration:

http://www.ana.unibe.ch/events/cemovis/index.html

Registration deadline: 15.04.2014

Yours faithfully

Benoît Zuber

Prof. Benoît Zuber
Institute of Anatomy
University of Bern
Baltzerstrasse 2
CH-3000 Bern 9
Switzerland
Tel. +41 31 631 84 40
benoit.zuber-at-ana.unibe.ch
http://www.ana.unibe.ch/~exmo/

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Hi Greg,

You might want to consider the New England Society for Microscopy (NESM
- http://www.nesmicroscopy.org/). They area a good bunch of people who
hold several meetings a year, typically in the eastern MA area.

Jeff

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Email: gr3g0rr33-at-gmail.com
Name: Gregory Wyche

Organization: University of South Carolina

Title-Subject: [Filtered] Networking Opportunities In CT

Message: Hi, I'm a microscopist in CT (currently between jobs) looking
for opportunities to network in that state. The Connecticut Microscopy
Society website has not been updated in a long time so I can't get in
touch with any of the members. If you know of any groups that meet,
please forward them. I'll very much appreciate it.

Login Host: 68.9.225.131
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Have you contacted

Derek Saunders
Electron Optic Services Inc
70 Bentley Ave; Suite 205
Ottawa, Ontario
K2E 6T8
CANADA
http://www.eos.ca
eos-at-eos.ca
(613) 723-7773
(819) 684-2308 fax

Ken Converse
owner

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

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Email: gul417-at-mail.usask.ca
Name: Guosheng Liu

Organization: University of Saskatchewan

Title-Subject: [Filtered] TEM (Philips CM10) service contract provider?

Message: Hello EM Community,

We are shopping for a service contractor for our CM10 TEM, ideally from
Canada or USA. This
20-year-old but sill reliable TEM has been service-contracted for most of
its years but run without
contract since early 2013(budge issue).....I was wondering if you would like
to share information
regarding service engineers/companies for the same or similar equipment. We
will purchase a service
coverage either very basic or all-inclusive depending on the quotes.

Thanks a lot.

Guosheng Liu

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Gregory,
Try the New England Society for Microscopy. They are very active.
www.nesmicroscopy.org

Ken Converse
owner

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

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Email: gr3g0rr33-at-gmail.com
Name: Gregory Wyche

Organization: University of South Carolina

Title-Subject: [Filtered] Networking Opportunities In CT

Message: Hi, I'm a microscopist in CT (currently between jobs) looking for
opportunities to network
in that state. The Connecticut Microscopy Society website has not been
updated in a long time so I
can't get in touch with any of the members. If you know of any groups that
meet, please forward
them. I'll very much appreciate it.

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Dear Listers,
if there's a hole in the window of an EDS detector mounted on a TEM,
what problem may arise using the detector?
I realizd that it's clear to me the window purpose in a SEM, where the
chamber is frequently vented, but what about in a TEM?
Thanks

Davide

~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~~~~~~~
Davide Cristofori

direct phone = 041.234.67.26
e-mail = dcristofori[at]unive[dot]it

Università Ca' Foscari Venezia
Dipartimento di Sc. Molecolari e Nanosistemi
Via Torino, 155b I-30172 Mestre (VE) Italy
~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~~~~~~~

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Hi Davide,

The purpose is to keep the detector crystal clean. Without the window
there, the detector will behave very much like a cold finger in any
other high or ultra high vacuum system. Residual water and other
volatiles in the system will condense on the crystal surface over time.
The effect may not be as immediate as it would be at atmospheric
pressure, but it can be just as devastating.

Windowless detectors can be used if the vacuum quality is very high (UHV
conditions with very little hydrocarbon contamination) or the detector
is relatively warm as with an SDD. Even with a windowless SDD, care
must be taken to ensure that the vacuum quality remains relatively high,
though UHV conditions are not required.

-Jacob

On 14-03-04 08:44 AM, dcristofori-at-unive.it wrote:
}
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} Dear Listers,
} if there's a hole in the window of an EDS detector mounted on a TEM,
} what problem may arise using the detector?
} I realizd that it's clear to me the window purpose in a SEM, where the
} chamber is frequently vented, but what about in a TEM?
} Thanks
}
} Davide
}
}
} ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~~~~~~~
} Davide Cristofori
}
} direct phone = 041.234.67.26
} e-mail = dcristofori[at]unive[dot]it
}
} Università Ca' Foscari Venezia
} Dipartimento di Sc. Molecolari e Nanosistemi
} Via Torino, 155b I-30172 Mestre (VE) Italy
} ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~~~~~~~
}
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--
Jacob Kabel
Electron Microscopist
Department of Materials Engineering
The University of British Columbia
6350 Stores Road, Room 419
Vancouver, British Columbia
Canada
V6T 1Z4
Phone: (604) 822-5648
Fax: (604) 822-3619
http://emlab.mtrl.ubc.ca/

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Dear Colleagues,

The program for the upcoming 3DEM GRC is mostly online now:
http://www.grc.org/programs.aspx?year=2014&program=threedim
This years's focus will be on "Technical Advances for a Rising Star in Structural Biology: 3DEM".
The site in Spain can house 200 guests. Over 100 participants are already registered, the conference is filling up quickly. The registration deadline is May 25. However, earlier registrations may be accepted sooner, in order to allow you to make travel arrangements in time.

Looking forward to meeting you in Spain for an exciting conference,

Henning.

Henning Stahlberg, PhD
Prof. for Structural Biology, C-CINA, Biozentrum, University Basel
Mattenstrasse 26 | D-BSSE | WRO-1058 | CH-4058 Basel | Switzerland
http://c-cina.org | Tel. +41-61-387 32 62 | mailto:Henning.Stahlberg-at-unibas.ch

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Hello,

EBSD 2014 is a topical conference sponsored by the Microanalysis Society (MAS) and will be held this June 17-19 at Carnegie Mellon University, Pittsburgh, PA, USA. This conference is a three-day event, highlighted by a one-day tutorial on the electron backscatter diffraction (EBSD) technique and two-days of technical symposia featuring invited and contributed talks on EBSD development and applications in materials science and geological studies.

Registration is currently open, but space is limited and registration slots are filling up quickly (deadline is March 31). Please see the link below for more information about the conference, to register, and to submit abstracts for oral/poster presentations (Abstract deadline is April 2):
http://www.microbeamanalysis.org/topical-conferences/EBSD-2014/welcome

Below is more details about the tutorial and technical symposia for EBSD 2014. We sincerely hope you can attend!

Tutorials - Tuesday, June 17th
Topics and speakers are listed below:
EBSD Technology (Des Moser and Andy Deal)
Sample Preparation (Brian Hess and Ron Witt)
Data Acquisition and Post Processing (David Mainprice)
Free, Open-source MTEX Data Processing Tools (David Mainprice)
Case Studies (David Mainprice and Joe Michael)

Live Demonstrations - Tuesday, June 17th
In parallel with the tutorial sessions, there will be opportunities to test out the latest and greatest EBSD systems from all major EBSD vendors (including Bruker, EDAX, Oxford Instruments, Thermo-Fisher).

Technical Symposia - Wednesday & Thursday, June 18th &19th
The EBSD 2014 technical symposia are the heart of the conference and consist of an exciting group of invited speakers at the forefront of EBSD technology and its applications. Below are the session topics and invited speakers headlining each session:

EBSD for Industry - Raj Mishra - General Motors, USA
EBSD and Energy Technologies - Helio Moutinho - National Renewable Energy Laboratory, USA
Microstructure Characterization - Stefan Zaefferer - Max Planck Institute, Germany
Computational Advances - Gert Nolze - BAM, Germany
Novel Methods - Pat Trimby - University of Sydney, Australia
Geochronology Phases - Steve Reddy - Curtin University, Australia
Deformation Fabrics - David Prior - University of Otago, New Zealand

Thank you for your interest and looking forward to seeing you at the conference this June!

EBSD 2014 Organizing Committee
David Fullwood - Brigham Young University
Elena Miranda - California State University - Northridge
Desmond Moser - The University of Western Ontario
Yoosuf Picard - Carnegie Mellon University
Andrew Deal - GE Global Research

John Mansfield PhD Cphys MInstP
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Davide

Just another comment. Yes the window serves
as a protective agent as outlined by others.

Although a TEM is not leaked in the same manner
as some SEM's there is still inevitably a small burst
of water vapor and gas that enters the column via the
insertion via the airlock. So there will be a gradual but slow build
up of condensate upon the cold detector, particularly
if it is an older SiLi system cooled by LN2.
Some older SiLi systems do have small heaters/thermal cycling
modes to mitigate ice build up. If you
have a SDD which is typically turned on/off then it
will warm up on power down and any ice will sublime
in vacuum.

What you don't want to happen is for the detector to be
fully powered up and the column to be leaked to air. Not only will
you get condensate, but more likely there will be a short
circuit of the electrical connections at the back of the
detector. The short circuit can destroy your detector.

Qualitative analysis will not be affected by a partially broken
window, unless you get ice build up from water vapor
in your column condensing on the front of the cold detector. This will
decrease the detection efficiency for light elements.
You will see this first on the low energy lines ( { 2 keV).

Please also remember since the presence or absence of
a window affects the relative intensities of x-ray lines, a change
in the window can impact any quantitative analyses.
Literally all the quantitative
analysis programs assume parameters for the initially
installed window and the manufacturers will have this
programmed into their analysis software.

A partially broken window will be difficult to model
as you don't know how much of the detector area
is affected. Depending upon the x-ray lines analyzed
this can change your analysis, again low eneryg lines
will be most affected.

So should you initially worry, I would not as long
as the detector is still working. I have run windowless
detectors (both LN2 cooled SiLi and SDD's) in TEM's for
years. With proper care and attention they are fine.

However, if you are in a multi-user facility with
variable expertise of users then I would play
it safe and get the window repaired. Fixing a window
is alot cheaper than replacing the whole crystal.

Nestor
Your Friendly Neighborhood SysOp

On 3/4/14 10:33 AM, dcristofori-at-unive.it wrote:
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} Dear Listers,
} if there's a hole in the window of an EDS detector mounted on a TEM,
} what problem may arise using the detector?
} I realizd that it's clear to me the window purpose in a SEM, where the
} chamber is frequently vented, but what about in a TEM?
} Thanks
}
} Davide
}
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} Davide Cristofori
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} direct phone = 041.234.67.26
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Title-Subject: [Filtered] silicone removal

Message: Can someone recommend a chemical recipe to remove silicone from an LED package device?

Thank you

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Hello Listservers,
I am planning a freeze substituition exp on cultured cells that have been
high pressure frozen for immunoelectron microscopy. I was wondering what
the experts use as a transition/dehydration solvent for IEM, cold (-90C to
-50c) methanol or acetone in their cocktail mixes. I have read where
methanol causes more tissue shrinkage (rt) and cold acetone is better,
although methanol may penetrate quicker. I am looking for acceptable
labeling and ultrastructure. Any advice is appreciated.

Sincerely,
Michael Delannoy
Assoc. Director Johns Hopkins SOM Microscope Facility
Baltimore, MD

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From rotoowy-at-vieuxportdemontreal.com Wed Mar 5 09:56:03 2014
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Marissa,

Silicones used in packaging of semiconductor devices are notoriously
difficult to remove, but:

1) Hot red fuming HNO3 or hot 3:1 mix of red fuming HNO3 with fuming
H2SO4 would work very nicely, if you do not mind loosing entire package
and be left with bare LED die.

2) Soaking in warm (40C) Methylene Chloride overnight (under a fume
hood) usually softens silicone to the point that it can be cleaned off
by a Q-tip. Sonicating in Methylene Chloride may be even better, but I
haven't try it. When using methylene chloride make sure it is dry - if
contaminated by water it becomes corrosive.

3) I had limited success with sonicating parts in warm Dynasolve 230
(under a fume hood) for a few hours.

Good luck :)

Valery Ray
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Dear All,
thank you for your replies and comments. They have been precious to me.
Just to answer some questions in your comments:
The detector is a Si(Li); we're going to repair the window (an upgrade
to SDD was considered, but we'll wait for the moment - sigh), just
wanted to have sounder idea of what would happen by operating the
detector if any user of our facility asked me for it.
Your answers clarified it.
Thanks again

Davide

~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~~~~~~~
Davide Cristofori

direct phone = 041.234.67.26
e-mail = dcristofori[at]unive[dot]it

Università Ca' Foscari Venezia
Dipartimento di Sc. Molecolari e Nanosistemi
Via Torino, 155b I-30172 Mestre (VE) Italy
~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~~~~~~~

Il 04/03/2014 17.39, dcristofori-at-unive.it ha scritto:
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} Dear Listers,
} if there's a hole in the window of an EDS detector mounted on a TEM,
} what problem may arise using the detector?
} I realizd that it's clear to me the window purpose in a SEM, where the
} chamber is frequently vented, but what about in a TEM?
} Thanks
}
} Davide
}
}
} ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~~~~~~~
} Davide Cristofori
}
} direct phone = 041.234.67.26
} e-mail = dcristofori[at]unive[dot]it
}
} Università Ca' Foscari Venezia
} Dipartimento di Sc. Molecolari e Nanosistemi
} Via Torino, 155b I-30172 Mestre (VE) Italy
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} 6, 20 -- From dcristofori-at-unive.it Tue Mar 4 10:33:15 2014
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From medications-popular7-at-xqqme.com Wed Mar 5 11:31:15 2014
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Dear List-Servers,

thank you for your suggestions and all your help! Sorry, that it took me so long to response. We were lucky and a FEI service-technician came this Monday. He found out that the problem was one of the valve, the NVC. So, currently I am working, quite happy, with stable 0.15 Torr (20 Pa) over the last 10h.

Thank you all again!
Stefan

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Email: levilr-at-ptd.net
Name: Lee Levine

Organization: Process Solutions Consulting

Title-Subject: [Filtered] LED Silicone removal

Message: I use a solvent SU100 by R Moreau. Not the cleanest removal but it is as good as anything
that I've found. It does swell the silicone possibly damaging wire bonds.

If you need to see the wire bonds do not use ultrasonic cleaners. They will make wire bonds fail in
seconds.

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Email: jcsmtf-at-mail.missouri.edu
Name: Josh Schorp

Organization: University of Missouri

Title-Subject: [Filtered] CM12 not staying on

Message: What could cause our CM12 to turn on but then shut off a few seconds later? This EM was
just re-assembled and I have just now been able to turn on the power. When I press the ON button it
sounds as if it turns on. After a few seconds it shuts off and will only turn back on if unplugged
then re-plugged back in. Does anyone have any insight on this?

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Josh,

I'm not sure of the exact problem but as a first test, check the power
supplies in the main power cabinet. They are behind the left door.
There is a 24 volt and a 5 volt supply (perhaps some 122 volt supplies
too). The power supplies have indicator LEDs on the front panel of the
supply. Make sure those are powering up.

Cheers,
Henk

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} Title-Subject: [Filtered] CM12 not staying on
}
} Message: What could cause our CM12 to turn on but then shut off a few seconds later? This EM was
} just re-assembled and I have just now been able to turn on the power. When I press the ON button it
} sounds as if it turns on. After a few seconds it shuts off and will only turn back on if unplugged
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From popular_medications13-at-achinsk.net Wed Mar 5 20:00:26 2014
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Hi, Dan

This looks like an interesting design, if you have an existing
spectrometer to slide it into.

Historically, I would have recommended the IlluminatIR, but it looks
like it is no longer made by Smiths Detection.

I am at Pittcon this week and saw a brand new system called Spero,
from Daylight Solutions (Google Daylight Solutions+Spero for their
information). It was exceedingly clever in its design... everything
was seamlessly integrated and the data output does not require highly
trained staff to analyze. They have a live imaging mode for
navigation and can collect a hyperspectral cube in record time.

Caveat: I have no commercial interest in any of these products.

Good hunting!
Barbara Foster, President & Chief Consultant
Microscopy/Microscopy Education*
www.MicroscopyEducation.com

*A subsidiary of The Microscopy & Imaging Place, Inc.
7101 Royal Glen Trail, Suite A
McKinney, TX 75070
P: 972-924-5310
F: 214-592-0277

MME is currently scheduling courses for now and through Fall 2014.
Call us today for a free training evaluation.

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

I can understand your skepticism, especially since I am a chemist by
training. However, I spent considerable time while writing an early
article on the IlluminatIR trying to figure out how to give
microscopists an easy way to analyze spectra. From the demo I had
today, these folks have solved that problem, at least with commonly
available spectra. Of course, my mentor in this area, John Reffner,
always advised collecting your own library, including the "sweepings
from the corner of the lab."

My suggestion: get a demo and decide for yourself.

Good hunting!
Barbara Foster, President & Chief Consultant
Microscopy/Microscopy Education*
www.MicroscopyEducation.com

*A subsidiary of The Microscopy & Imaging Place, Inc.
7101 Royal Glen Trail, Suite A
McKinney, TX 75070
P: 972-924-5310
F: 214-592-0277

MME is currently scheduling courses for now and through Fall 2014.
Call us today for a free training evaluation.

At 10:48 PM 3/5/2014, you wrote:
} Re: "...the data output does not require highly
} trained staff to analyze."
}
} Uh huh, sure.
}
}
} -----Original Message-----
} } From: bfoster-at-the-mip.com
} } Sent: Mar 5, 2014 11:06 PM
} } To: jtwilley-at-sprynet.com
} } Subject: [Microscopy] Fwd: viaWWW:FTIR Microscope
} }
} }
} }
} }
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} }
} } Hi, Dan
} }
} } This looks like an interesting design, if you have an existing
} } spectrometer to slide it into.
} }
} } Historically, I would have recommended the IlluminatIR, but it looks
} } like it is no longer made by Smiths Detection.
} }
} } I am at Pittcon this week and saw a brand new system called Spero,
} } from Daylight Solutions (Google Daylight Solutions+Spero for their
} } information). It was exceedingly clever in its design... everything
} } was seamlessly integrated and the data output does not require highly
} } trained staff to analyze. They have a live imaging mode for
} } navigation and can collect a hyperspectral cube in record time.
} }
} } Caveat: I have no commercial interest in any of these products.
} }
} } Good hunting!
} } Barbara Foster, President & Chief Consultant
} } Microscopy/Microscopy Education*
} } www.MicroscopyEducation.com
} }
} } *A subsidiary of The Microscopy & Imaging Place, Inc.
} } 7101 Royal Glen Trail, Suite A
} } McKinney, TX 75070
} } P: 972-924-5310
} } F: 214-592-0277
} }
} } MME is currently scheduling courses for now and through Fall 2014.
} } Call us today for a free training evaluation.
} }
} }
} }
} }
} } } Envelope-to: bfoster-at-the-mip.com
} } } Date: Mon, 3 Mar 2014 19:10:58 -0600
} } } To: bfoster-at-the-mip.com
} } } From: microscopylistserver-noreply-at-microscopy.com
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} } } Name: Daniel VanHart
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} } } Title-Subject: [Filtered] FTIR Microscope
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} } } Message: Does anyone have any experience with a Pike Technologies
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From medications-canadian9-at-nationalcablenetworks.ru Thu Mar 6 00:22:58 2014
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I've recently found a very nice parameterization of K-shell electron-impact ionization cross-sections (Talukder et al., Int. J. Mass Spec., V269 (2008) P. 118 and V309 (2012) P. 212) which are very convenient to code into the computer.

I'm not finding similar good (or even fair) parameterizations for L- or M-shells. Can anyone recommend good references from which to write code for X-ray cross-sections?

Thanks
Chad

---------------------
Chad M. Parish, Ph.D.
Research and Development Staff Member
Materials Science and Technology Division
Oak Ridge National Laboratory
Phone: 1 865 574 0092
Email: parishcm-at-ornl.gov

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Hi all,

We have a two-year postdoc position in atomic resolution STEM imaging
(experiment, theory or both) at Monash University, Australia. The
position is in collaboration with the Monash Centre for Electron
Microscopy (inc. double-corrected Titan) and the theory group of Prof.
Les Allen at the University of Melbourne. Further details below.

Please bring this position to the attention of anyone who might be
interested.

Many thanks,
Scott Findlay

________________________________________________________

Position description and application details available at:
http://jobs.monash.edu.au/jobDetails.asp?sJobIDs=521173&lWorkTypeID=&lLocationID=&sJobNo=electron&lCategoryID=&lBrandID=&sKeywords=electron&stp=AW&sLanguage=en.

Position overview: We seek a postdoctoral researcher to conduct research
in atomic resolution STEM imaging and spectroscopy. The postdoctoral
researcher will work with Dr S.D. Findlay, A/Prof. M. Weyland, Dr A.J.
D’Alfonso and Prof. L.J. Allen on atomic resolution imaging and
spectroscopy on a project funded by the Australian Research Council. The
position will be formally located in the School of Physics, Monash
University, but will involve extensive collaboration with the Monash
Centre for Electron Microscopy, and the Theoretical Condensed Matter
Physics group at the University of Melbourne. The project will also
involve collaborative work with colleagues in the UK, USA, Europe and
Japan. The postdoctoral researcher will conduct research in experimental
and/or theoretical aspects of atomic resolution electron microscopy at
the highest international levels and translate research outcomes into
high impact publications, present results at major conferences and
assist in the supervision of PhD and final year undergraduate students.

Duration: Two-year appointment

Remuneration package: $79,485 - $85,323 pa Level A PhD / $91,609 -
$108,788 pa Level B
(inc 9.25% employer superannuation)

Closing date: Monday 31st March, 2014

Job No: 521173

Questions regarding the position can be directed to Dr Scott D. Findlay,
scott.findlay-at-monash.edu, A/Prof. Matthew Weyland,
matthew.weyland-at-monash.edu, Dr Adrian J. D'Alfonso,
adrian-at-dalfonso.com.au, or Prof. Leslie J. Allen, lja-at-unimelb.edu.au.

--
Dr Scott Findlay
QEII Fellow
School of Physics
Monash University
Victoria, 3800, Australia
Tel: +61 3 9902 4943
Fax: +61 3 9905 3637

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Email: robernal-at-u.northwestern.edu
Name: Rodrigo Bernal

Organization: Northwestern University

Title-Subject: [Filtered] Case for JEOL TEM Specimen Holder

Message: Does anyone know any vendor where I could purchase this case for a single tilt holder?

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From meds-popular14-at-starlink.ru Fri Mar 7 06:27:28 2014
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Hello Listservers,
What is the storage life of LN2 frozen AFS cocktails (0.2% glutaraldehyde,
0.25% uranyl acetate in 95% acetone) for HM20 IEM? Given that the glut
stocks (10% acetone) when stored at 4C have a shelf life of 2 years.

Sincerely,
Michael Delannoy

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Rachel,
The original stocks of 10% Ga in acetone where never opened (we use so little from each ampoule), I assume sealed under nitrogen. This solution polymerized in the bottles within 2 years (EMS confirmed 2 year shelf life at 4C). The working solutions (0.2% Ga 0.2% UA 95% acetone) were frozen and stored under LN2 conditions (-200C). I agree with you in the addition of 2-5% water to visualize membranes (bi-layer), and am curious about your IEM with 0.1% osmium. Do these sections need to be etched (saturated sodium-m-periodate) before labeling?
Do you have a Ga Ua Osmium cocktail? What are the percentages?

Sincerely,
Michael Delannoy

-----Original Message-----
X-from: Reinhard Rachel [mailto:Reinhard.Rachel-at-biologie.uni-regensburg.de]
Sent: Friday, March 07, 2014 10:50 AM
To: Michael Delannoy

-----Original Message-----
X-from: Reinhard Rachel [mailto:Reinhard.Rachel-at-biologie.uni-regensburg.de]
Sent: Friday, March 07, 2014 10:50 AM
To: mdelann1-at-jhmi.edu

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Title-Subject: [Filtered] Double Tilting Motor Electrical Diagram

Message: Hi,

The double tilt holders for JEOL usually have a standard connector that plugs into a terminal in the
goniometer. In this way, one can control the secondary tilt from the microscope software. I am
designing a custom double tilt holder and hope to make it motorized. Does anyone have the electrical
diagram of this connector so that I can know how to wire my holder properly?

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Email: tprozoro-at-ameslab.gov
Name: Tanya Prozorov

Organization: US DOE Ames Laboratory

Title-Subject: [Filtered] Faraday cup holder for Tecnai G2 F20

Message: Dear Listers,

I was wondering if someone has a Faraday Cup holder compatible Tecnai G2 F20 for a direct
measurement of current on a specimen in STEM mode.
We need to read the electron dose and dose rate, and our estimates seem to be off quite a bit.

If you know someone who knows someone, I would very much appreciate if you can refer me to this
person. If you are the lucky owner, please contact me to discuss whether this could be arranged.

Many thanks,

-Tanya
---------------------------------t--------------------------
Tanya Prozorov, Ph.D.

Emergent Atomic and Magnetic Structures
Division of Materials Sciences and Engineering
US DOE Ames Laboratory
332 Wilhelm Hall, Ames IA 50011
515-2943376 (office)
515-2941255 (lab)
515-294-8727 (fax)
e-mail: tprozoro-at-ameslab.gov
CREATING MATERIALS AND ENERGY SOLUTIONS

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Email: jcsmtf-at-mail.missouri.edu
Name: Josh Schorp

Organization: University of Missouri

Title-Subject: [Filtered] CM12 vacuum

Message: Our CM12 was at atmospheric pressure before turning on. When turned on, the prevac pumps
down to P2=30. Then V10 will open, P2 will rise and slowly fall back down to 30. V2, V7, V5, V6, and
ODP are all highlighted on the the screen. P1=35 and P3 will only go down to 66 after about 2 hours.
The prevac turns on every 5 minutes or so and lowers P2 back down to 30 then shutting off, but once
turned off, P2 goes up to 80. Is this correct, or do we have a problem with the ODP, leaks, etc.?

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Email: john.kourtesis-at-sars.uib.no
Name: John

Organization: uniSAS, Bergen, norway

Title-Subject: [Filtered] LR White sectioning

Message: I have been facing difficulties in sectioning long consistent ribbons of 100nm thin
sections of LR White biological tissue. Although I lower a lot the water on the trough and I use
antistatic gun, the sections tend to submerge in the water

any hints, suggestions for more efficient ribbon sectioning?

many thanks

John

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From meds-cheapest8-at-mitchellcompanies.com Sat Mar 8 21:38:39 2014
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Hi All,

Our software is not showing any x-ray counts and can't collect a
spectrum when a sample is scanned.

We have a Bruker XFlash SDD detector set up on our JEOL SEM, with each
device connected to it's own PC.

The PC's are connected to our network for internet access. Our IT
department came through our lab the other day to address a network
connectivity issue. After that, the Bruker Espirit software wasn't
able to detect our X-Ray detector, an XFlash 4010. Turns out an
ethernet cable going from the electronics signal processing box to the
pc had been pulled. But even after reconnecting the software couldn't
connect to the detector. We swapped cables, still no good. After a few
days, we realized that the cable was a cross-patch cable, so we tried
an ethernet cable out of the wall, and voila it reconnected to the
detector and was getting counts.

But then after a session where it got reasonable counts, that matched
previous scans of the same sample, the next day, no counts. The
detector is recognized, Espirit can read some data from it (like the
temperature of the detector), but it wouldn't register any counts.

We've checked the connections many times, especially because it looks
like IT just pulled everything that looked like an ethernet cable. The
Detector is connected to it's electronics box with a megalink cable,
which seems securely in place, the electronics box is lighting up and
indicating that data is being transferred, and the box is connected to
the PC through a card which seems to be working properly otherwise (it
accepts video from the SEM and that is picked up perfectly by the
Espirit software, including operating conditions information, like
voltage and magnification).

So it doesn't sound like there's any connection problem, but I have to
wonder if the ethernet cable we've pulled out of the wall is really
the same as what might be normally used for the electronics box to PC
connection?
They're both listed as Cat5e cables.

We've selected different throughput cards through the software, but
still no counts. The detector hasn't been moved out of position, isn't
touching the pole piece, and nothing seems to be blocking it. Bruker
wants us to send in the detector and box, but they think it's likely
that the detector itself is dead.

Has anyone had a similar problem and/or knows a resolution?

Our detector is around 7 years old, does that sound like a reasonable
lifetime for a detector?

--
Robert J. Schenck
Kingsborough Community College
Physical Sciences Department
Follow Me on Twitter: -at-Schenck

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11, 26 -- Subject: [Microscopy] Bruker XFlash SDD X-ray detector no longer getting any counts
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Dear colleagues - my group is presently looking for postdoctoral
candidates in the following areas:
1. In-situ oxide growth/atomically resolved STM/surface science
characterization of fundamental physical and chemical phenomena on oxide
surfaces
2. Dynamic SPM methods (multifrequency/band excitation)
3. Mesoscopic characterization of ferroelectric and energy storage and
conversion materials
All positions will utilize unique instrumental platforms developed at
CNMS and will allow for significant independent program building. Please
contact me directly for additional details.
Sergei Kalinin

--
Sergei V. Kalinin

New:
CNMS on FaceBook: https://www.facebook.com/CNMSORNL#!/CNMSORNL
Google Scholar: http://scholar.google.com/citations?user=-cuxoSQAAAAJ&hl=en

Theme Leader for Electronic and Ionic Functionality
The Center for Nanophase Materials Sciences
and Materials Sciences and Technology Division
Oak Ridge National Laboratory
Oak Ridge, TN 37831

Joint Faculty Associate Professor,
Bredesen Center for Interdisciplinary Research and Education
University of Tennessee, Knoxville
http://bredesencenter.utk.edu

Phone: (865) 241-0236
http://www.cnms.ornl.gov

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From gkojaje-at-cds-engineering.com Tue Mar 11 02:13:51 2014
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FYI

Inter/Micro 2014
June 2-6
at McCrone Research Institute in Chicago, Illinois
McCrone Research Institute cordially invites you to participate in
Inter/Micro 2014.

http://www.mcri.org/home/section/101-915/inter-micro-2014

Papers are being solicited in micro-analytical techniques and
instrumentation, environmental and industrial microscopy, and chemical and
forensic microscopy.

The deadline to submit titles and abstracts is April 1, 2014.

For more information, visit our website at: www.mcri.org

Contact us at (312) 842-7100 or by email at: intermicro-at-mcri.org

We look forward to seeing you at Inter/Micro in Chicago!

McCrone Research Institute | 2820 S. Michigan Avenue | Chicago | IL | 60616

Tony
.........................................
Andrew Anthony "Tony" Havics, CIH, PE
Environmental, Health & Safety, Microscopy, Materials Science & Forensic
Engineering
pH2, LLC
5250 E US Highway 36, Suite 830
Avon, IN 46123
(317) 718-7020 Office
(317) 718-7038 Fax
(317) 409-3238 Cell
aahavics-at-pH2LLC.com
www.ph2LLC.com

This message is from pH2. This message and any attachments may contain
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Email: sgkcck-at-aol.com
Name: Stacie Kirsch

Organization: Electron Microscopy Sciences

Title-Subject: [Filtered] Webinar - Datacolor CHROMACAL

Message: Hello everyone,

Just wanted to let you know about an upcoming webinar I invite you to join! Here are the details:

What: Webinar - Ensuring Color Integrity with Datacolor CHROMACAL - Learn about the Features and
Benefits of this new product!
When: Wednesday, April 2, 2014 1:00 PM Â– 2:00 PM EDT
Who: Datacolor and Electron Microscopy Sciences
Where to register: https://www2.gotomeeting.com/register/244965202
Additional information:
Online Flyer: http://www.emsdiasum.com/microscopy/emailnews/chromacal_mar2014.html
Website: http://www.emsdiasum.com/microscopy/products/calibration/systems.aspx or
http://www.emsdiasum.com/chromacal
Cost: free

Let me know if you have any questions! Thanks

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

FYI

Dear Colleagues,
I should be glad if there are some dermatologist and skin researchers using the Microscopy Listserver who might find this information useful:

Announcement
41st Annual Meeting of the SCUR (Society for Cutaneous Ultrastructure Research ROME,ITALY; 2014)
June 5-7,2014

1st Announcement 41st Annual Meeting of the SCUR
(http://www.scur.org/content/e1174/e1181/e1182/index_ger.html)

"VILLA MONDRAGONE", Monte Porzio Catone (RM, ITALY) JUNE 5-7, 2014
GENERAL TOPIC: "SKIN AGEING and CANCER: From ULTRASTRUCTURE to CLINIC"
INFORMATION (Flyer all forms)

Call for papers (in dermatology, dermatopathology, skin research,
presenting of results from all ancient and modern imaging techniques)

Registration and Accommodation information available.

The deadline to submit titles and abstracts is March 20th, 2014.
(late entries will be accepted on demand and need to be indicated by a mail to the
Local Organizers at
costanza-at-med.uniroma2.it; anapat-at-uniroma2.it; a.orlandi-at-uniroma2.it )

For more information, visit our website at:
http://www.scur.org/content/e1163/e1164/index_ger.html
(www.scur.org)

Contact us (in case of requests for special information)by email at:
also: W.Muss-at-salk.at

We look forward to seeing you at the 41st Ann. Meeting of the SCUR
in Monte Porzio Catone (RM), 20 km southeast of ROME city center
(FRASCATI, near airport CIAMPINO or FIUMICINO), a marvelous place to stay!

Wolfgang Muss, PhD
.........................................
SCUR Secretary
Wolfgang MUSS
EM-Lab, Univ. Inst. Pathology,
SALK-LK (Gen. Hospital) and PMU (Private Paracelsus Medical University) Salzburg
A-5020 SALZBURG-AUSTRIA

==============================Original Headers==============================
18, 38 -- From W.Muss-at-salk.at Wed Mar 12 09:24:23 2014
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18, 38 -- Ultrastructure Research ROME,ITALY; June 5-7,2014
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18, 38 -- Ultrastructure Research ROME,ITALY; June 5-7,2014
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From drugs-cheapest4-at-quicktel.com Wed Mar 12 10:05:59 2014
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For those who are interested:

Many months ago I posted on list server asking for feed back as to
which slow scan image capture system was the best. Thanks to all those
who responded to both my enquiry and to past posts for the useful
titbits. I should add I do not have shares in the company but am just
a very satisfied customer. There was not much on these products on
list server so I though I would just mention our deliberations.

Our short list was boiled down to 2 contenders either the quartz PCI
system or the Saturn Imaging System (E.L.I. Orionmicroscopy.com)
mainly I have to say based largely on cost.

For our users the main priority was ease of image capture and quality.
Many of the other image capture systems we considered (not mentioned)
put an over emphisis on post processing. We liked the simplcity of
these two systems. Our users pay by the hour and do all the post
processing on standalone workstations away from the microscope using
open source or sitle licenced software.

We decided on the Saturn as it was not only very competively priced
but offered active scan control of our FESEM (Jeol 6301F) which allows
for much higher resolution scan than the default passive sync could
allow (although it does have the passive modes also). It also allows
for much clearer and noise free images. The layout is very simple and
uncluttered and it allows the users to easily drive the SEM without
fuss.

Given the age of the SEM it has given the machine a new lease of life
and we have had no complaints from our varied users. We previously had
a donated ADDAII which did not even come close with image quality.

In terms of PC it is a win7 box with no difficult cards inside so
future maintainence should not be an issue. The electronics controller
for the active scan is tiny and tucks away out of sight.

Jean Louis Leclef the proprieter designs, manufatures and installs the
units himself and he was really optimised the system for us. We have
ended up with a bespoke system that meets our requirements exactly. He
even changes the software buttons to read out the way that suits the
instrument.

Several desirable features have been incorportated for us which we
find invaluable. In addition to the standard calibration linearisation
methods he has implemented a method of calibrating images at all
magnifications, probe currents, accelerating voltages, working
distances etc so we can produce more accurate values for measuements.
Our sem being vintage suffers from non linearilities and so this
allows us to make very accurate measuments up to 500kx. He also has
addaped the software to include a waveform display for live
contrast/brightness adjustment and the system can do 0.2sec scans so
you get an almost TV rate image on the PC. This is good for us as we
have CRT screens on the SEM, these can be a bit noisy and weak with
the Cold FEG.

The quality of the images is supurb, we have had no problems getting
reliable publication quality images, from even the most inexperienced
student. Users find the simple measurement tools and the way it stores
data very conventient. All measurements are automatically exported
with calibrated text files for imediate use, there is no need to batch
export any more.

Jean Louis has given us great service support, every thing we wanted
the software to do he has impliemented. We have had no problem with
post instalation problems, these have been rectified within hours in
most cases (most of these related to software alterations that we
wanted not the hardware).

We aslo found this system was suitable to our STEM-TEM (Jeol 1200mk2)
but we decided to upgrade the EDS system with image capture instead of
this.

Should anyone require any futher information, images or comments
please feel free to contact me off list. Like I say, I am not a sales
rep just a very happy customer.

John Mitchels
University of Bath, UK

==============================Original Headers==============================
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14, 26 -- Subject: Review of Saturn Active Image Capture System
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From meds-popular7-at-promtehnn.ru Thu Mar 13 13:11:30 2014
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Message-Id: {201403131811.s2DIBPv2030473-at-ns.microscopy.com}

Dear Listers,

For those of you doing color brightfield imaging, color integrity in
microscopy will be part of MBL's Immunohistochemistry and Microscopy
Workshop (March 15-20, Woods Hole, MA). Datacolor will have
scientific staff supporting the workshop, discussing current trends
in color integrity for microscopy and guiding hands-on lab exercises
using the ChromaCal microscope calibration system to standardize
color brightfield images and calibrate monitors for consistent,
reliable color image viewing. For further details and a link to the
workshop, visit http://scientific.datacolor.com/resources/events/

Best regards,
Barbara Foster, President & Chief Consultant
Microscopy/Microscopy Education*
www.MicroscopyEducation.com

*A subsidiary of The Microscopy & Imaging Place, Inc.
7101 Royal Glen Trail, Suite A
McKinney, TX 75070
P: 972-924-5310
F: 214-592-0277

MME is currently scheduling courses for now and through Fall 2014.
Call us today for a free training evaluation.

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

Hi

We are interested in a short course that focuses on Electron Microscopy for cellular biology. Is there such a course? We have been to Lehigh but that focuses on Materials Science. We are particularly interested in sample preparation techniques: proper fixation, staining, microtome and then Cryo TEM to examine the samples.

Thanks,

Pete Eschbach
Electron Microscope Facility Director
541 737 5645

Sent from my iPhone

==============================Original Headers==============================
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5, 32 -- Subject: EM techniques for cellular biology training?
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Email: cgoldsmith-at-cdc.gov
Name: Cynthia Goldsmith

Organization: Southeastern Microscopy Society

Title-Subject: [Filtered] 50th anniversary meeting of the Southeastern Microscopy Society

Message: Join us in Columbia, SC, May 20 through 23 for the 2014 meeting of the Southeastern
Microscopy Society! This will be the 50th Anniversary for the SEMS meeting and we are planning a
large celebration and hope to see many friends and past members of SEMS, as well as welcoming many
new members. Jay Jerome is the Program Chair for the meeting and many exciting talks and a strong
Ruska competition for our students are being planned. Bob Price is the local arrangements chair and
will be hosting a pre-meeting reception/fish fry at his home on the evening of May 20. All meeting
participants are invited to attend.

The meeting will be held at the Clarion Hotel which is located at 1615 Gervais St in the downtown
area of Columbia, SC. Visit our website at http://southeasternmicroscopy.org/2014-meeting for more
information and a link to a 2-page flyer that will give additional information.

Please mark your calendars now for our 50th Anniversary meeting and help us make this milestone in
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All,

We acquired a second-hand Robinson BSD that came with no paperwork. Can someone help me with a copy of the installation guide, operators manual and schematics for an older Robinson detector. I'm happy to pay copy costs and shipping.

Thanks, Owen

Owen P. Mills
Director, Materials Characterization & Fabrication Facilities

Materials Science & Engineering
Michigan Technological University
1400 Townsend Drive
Houghton, MI 49931
PH 906-369-1875
FAX 906-487-2934

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Colleagues

Just a short note that the Microscopy Listserver is back on-line.

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I need a little help with setting up my X-ray detector in Gatanâ€™s Digital Micrograph DigiScan and DTSA-II programs. I have a JEOL 2100F with a High Resolution pole piece and with an Oxford Inca X-sight detector. Oxford would not give me the detector parameters that the programs need citing proprietary information. They sent me a Moxtek data sheet, but didnâ€™t tell me which window it has, the AP3.3 or the AP3.7. Regardless, that information doesnâ€™t help anyway, because Moxtek doesnâ€™t give the parameters that the programs are asking for either. From my Oxford data sheet, it says that I have an ATW2 window.

Below are the parameters that I put into Digital Micrograph and DTSA. If you know the correct values or can help me figure out what they should be, please help. I don't know what the definition of the "zero fwhm" value is for the Gatan software is at all.

Geometry:
Elevation angle: 19Â° (18.7Â°)
Azimutahal angle: 270Â°
Incidence angle: 90Â°
Stage tilt: 0Â°
Solid angle 0.0616 sRad

Detector:
Dead layer: 100 nm (best guess)
Active layer: 3e+006 nm (best guess is 3mm thick)
Gold layer: 20 nm (best guess)
Window thickness: 100 nm (Best guess)
Window Type: Pyrolene (Best guess â€“the other choices are Aluminum, Boron Nitride, Diamond, and Hydrocarbon)
Detector type: Si(Li) (finally, one I know for sure)

Characteristic Physics:
Zero fwhm: 0.08049 (This value was already in DM EDS setup and I have no idea what should be or what it is. Anybody know?
Fano: 0.1132 (This value was already in DM EDS setup, the theoretical value for Si is 0.115 from my search)

Here are the parameters that I used for DTSA:
Window: Moxtek AP3.3 (It was the only Moxtek window definition from the data sheet from Oxford that I had)
Elevation angle: 18.7Â°
Azimuthal angle: 270Â° (I am taking the back of the TEM rod as the positive X-axis, i.e., 0Â°.)
Optimal working distance: 6.39 mm (I took the sample-to-detector distance *sin(18.7Â°)
Sample-to-detector distance: 19.927 mm (from the Oxford drawing sheet for the HR pole piece)

Crystal Parameters:
Detector area:30 mm^2 (known)
Gold layer: 8 nm (This is DTSA default value. Good as any that I know of.)
Dead Layer: 0.085 (This is DTSA default value. Good as any that I know of.)
Thickness: 3mm (The default for DTSA was 5, but I chose 3 mm. The intensity of the higher energy peaks on this detector seem to be low so I chose the lower number. Again, Best guess.)
Resolution: 129 eV (This was recently measured by Oxford on a PM visit.)

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I wanted to share a posting by John Oreopoulos to the confocal listserv. Tom Phillips

Begin forwarded message:
X-from: John Oreopoulos {john.oreopoulos-at-UTORONTO.CA}

Tom -

For a 36 page detailed PDF about this scope, see
http://arxiv.org/abs/1403.1211 The 50 cent cost of the scope is
based on the cost of components for 10,000 of them.

Caroline
--
Caroline Schooley
Project MICRO Co-Chair
Microscopy Society of America
45301 Caspar Point Road, Box 117
Caspar, CA 95420
Phone/FAX (707)964-9460
Project MICRO: http://www.microscopy.org/education/projectMICRO

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Dear Listers,

In my previous post, part of my question asked what the term "zero fwhm" term was in the Gatan Digital Micrograph software. Â

I'd like to verify it is essentially the Noise value in the following expression for the resolution at a given X-ray energy,

FWHM = SQRT[ Noise^2 + 2.35^2 * epsilon * F & E], where epsilon is 3.8 eV for a Si detector, F is the Fano factor, ~0.115, and E is the energy of the X-ray. Â For my detector, the resolution for Mn-Ka was measured to be 129 eV, so I calculated the "Noise" value to be 49 eV. Â I assume that this is the correct number to put into the Gatan software.

Please comment if I am incorrect.

Thanks.

-Scott Walck

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Email: svora-at-iic.cas.cz
Name: Petr Svora

Organization: Institute of Inorganic Chemistry, AS CR

Title-Subject: [Filtered] PECs component, part Gatan-Model 682.06651

Message: This Coating/Etching High Switching Module stop work in our PECs. Do somebody have similar
experience with it? Is it possible to repair it? Or somebody have unuse PECs :-)?

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From medications-discounted2-at-mgts.by Mon Mar 17 07:28:20 2014
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Scott;
Someone from Oxford once told me that there is no gold layer on their detectors-they use nickel.

John Mardinly, ASU

I need a little help with setting up my X-ray detector in Gatanâ€™s Digital Micrograph DigiScan and DTSA-II programs. I have a JEOL 2100F with a High Resolution pole piece and with an Oxford Inca X-sight detector. Oxford would not give me the detector parameters that the programs need citing proprietary information. They sent me a Moxtek data sheet, but didnâ€™t tell me which window it has, the AP3.3 or the AP3.7. Regardless, that information doesnâ€™t help anyway, because Moxtek doesnâ€™t give the parameters that the programs are asking for either. From my Oxford data sheet, it says that I have an ATW2 window.

Below are the parameters that I put into Digital Micrograph and DTSA. If you know the correct values or can help me figure out what they should be, please help. I don't know what the definition of the "zero fwhm" value is for the Gatan software is at all.

Geometry:
Elevation angle: 19Â° (18.7Â°)
Azimutahal angle: 270Â°
Incidence angle: 90Â°
Stage tilt: 0Â°
Solid angle 0.0616 sRad

Detector:
Dead layer: 100 nm (best guess)
Active layer: 3e+006 nm (best guess is 3mm thick) Gold layer: 20 nm (best guess) Window thickness: 100 nm (Best guess) Window Type: Pyrolene (Best guess â€“the other choices are Aluminum, Boron Nitride, Diamond, and Hydrocarbon) Detector type: Si(Li) (finally, one I know for sure)

Characteristic Physics:
Zero fwhm: 0.08049 (This value was already in DM EDS setup and I have no idea what should be or what it is. Anybody know?
Fano: 0.1132 (This value was already in DM EDS setup, the theoretical value for Si is 0.115 from my search)

Here are the parameters that I used for DTSA:
Window: Moxtek AP3.3 (It was the only Moxtek window definition from the data sheet from Oxford that I had) Elevation angle: 18.7Â° Azimuthal angle: 270Â° (I am taking the back of the TEM rod as the positive X-axis, i.e., 0Â°.) Optimal working distance: 6.39 mm (I took the sample-to-detector distance *sin(18.7Â°) Sample-to-detector distance: 19.927 mm (from the Oxford drawing sheet for the HR pole piece)

Crystal Parameters:
Detector area:30 mm^2 (known)
Gold layer: 8 nm (This is DTSA default value. Good as any that I know of.) Dead Layer: 0.085 (This is DTSA default value. Good as any that I know of.)
Thickness: 3mm (The default for DTSA was 5, but I chose 3 mm. The intensity of the higher energy peaks on this detector seem to be low so I chose the lower number. Again, Best guess.)
Resolution: 129 eV (This was recently measured by Oxford on a PM visit.)

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From meds_discounted5-at-ramnet.su Mon Mar 17 21:43:57 2014
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Greetings All,

The Midwest Microscopy and Microanalysis Society will hold its Spring meeting on Friday, March 28, 2014, at Northwestern University in Evanston, IL. The topic is Innovative Biological Microscopy, and details can be found on our website under Meetings:

http://www.midwestmicroscopy.org/

Please join us for an interesting and informative program and a chance to meet with your local microscopy colleagues. We look forward to seeing you there!

If you visit our website, you'll also find the latest edition of our newsletter, with a review of last year's meetings, 2014 meeting dates, links to our new social media sites and information about our 2014 Student Travel Awards for attendance at M&M 2014/IUMAS-6 in Hartford, CT.

Best regards,

Elaine Schumacher
M3S Program Coordinator

*********************************************************************
Elaine F. 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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Name: Andre Dufresne

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Title-Subject: [Filtered] Hitachi TM 1000

Message: Hi Everyone, The TM 1000 has been around for a while now and
we have had our Turbo Molecular Pump (TMP) refurbished and replaced it
ourselves in the past. We now have to repeat the process however the
availability locate a contractor that will refurbish our non
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Title-Subject: [Filtered] SEM aperture

Message: Dear friends,

Today I observed some biological samples, prepared normally with the
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However, when I returned to the Auriga/Zeiss microscope after few hours,
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Hello Daniela,
when this happens to me (using old JEOL SEMS) it is likely that the beam axis has shifted.
1. you see noise = the PMT works fine
2. you see a picture using other apertures (I assume they have larger diameters)= the beam is nearly ok and the PMT works well.

conclusion: a) either the beam is shifted a few µm, or the aperture foil has moved, or the GUN shift and tilt controls on the control board has been altered. b) The aperture could also have been blocked or degraded by the beam.
solution: a) adjust the GUN shift and tilt for maximum signal or adjust the µm knobs on the aperture holder. b) change aperture foil or clean it.
Best regards
Erik Ordell

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From pharmacy-best10-at-windelsmarx.com Fri Mar 21 02:18:48 2014
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I've been microtoming low Tg polymer recently and my cryo-microtome has the options of having the sample and knife at different temperatures.

How low? Well I'm working at -100C with a dry diamond knife and picking up the thin, crinkled sections with sugar water. I am not happy with the thin sections and I'm wonder if there is any advantage in having the knife and sample at different temperatures. It's an RCM Powertome with cryo attachment.

Any thoughts and experience I can draw on?

Thanks
Frank Karl

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Hi Daniela,

} When this happens to me (using old JEOL SEMS) it is likely that the beam axis has shifted.

This sounds like the case. The gun alignment is so far out that you can
no longer get an image.

The Zeiss Auriga, working on the assumption that it's a Zeiss Gemini
column, uses an intermediate lens to change apertures, so all of the
adjustments are electronic. The first thing to check is the emission
image on the 30um aperture. If your software is anything like mine, it
will be be a button labelled "Emission" under the apertures tab. This
will start the system rastering the gun alignment coils across the fixed
aperture plate. On my system the scanning has to be set to speed 9, and
then the "Gun Align" controls on the aperture tab can be used to adjust
the gun such that the selected aperture is in the centre of the image.

In the emission image, you should be able to see all of the available
apertures laid out in a circle. If the 30um aperture is not visible at
all in the emission image (probably the centre aperture, depending on
configuration) then you have bigger problems and a service call may be
required. From what I understand, the gun has to be removed to access
the aperture plate for replacement or cleaning.

If you don't have access to the gun alignment, or if it is so far out as
to be uncorrectable, you'll have to log in to the system with a service
level account to align it further. The user level accounts on Zeiss
seem to apply a limited offset to the major alignment performed by the
service engineers.

-Jacob

On 14-03-20 10:54 PM, erikordell-at-icloud.com wrote:
}
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} Hello Daniela,
} when this happens to me (using old JEOL SEMS) it is likely that the beam axis has shifted.
} 1. you see noise = the PMT works fine
} 2. you see a picture using other apertures (I assume they have larger diameters)= the beam is nearly ok and the PMT works well.
}
} conclusion: a) either the beam is shifted a few µm, or the aperture foil has moved, or the GUN shift and tilt controls on the control board has been altered. b) The aperture could also have been blocked or degraded by the beam.
} solution: a) adjust the GUN shift and tilt for maximum signal or adjust the µm knobs on the aperture holder. b) change aperture foil or clean it.
} Best regards
} Erik Ordell
}
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Jacob Kabel
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Canada
V6T 1Z4
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Title-Subject: [Filtered] amber on sem

Message: hi ,can i study amber under the scanning electron microscope in high vacuum mode.

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Good Day: Â We are doing high pressure freezing and freeze substitution on plant, animal and bacterial samples. We are using Epon resin for embedding but struggle with its viscosity. Our experience with Spurr's has been disappointing in that we get no or reverse contrast in the TEM. We love the ease of use but can't make it work for imaging.
Â Â We are using recipes from Glauert (firm and the kit) and have both a new kit and older stocks of the original ingredients. It doesn't seem to make a difference. Â
Â Â Does anyone have any advice on how we can make and use this resin to produce samples that produce publication quality contrast in our images? Thanks bob

--
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Interesting question...
I suspect charging will be a major problem, so I would suggest a gold coating and a little extra effort to make sure the sample is grounded. I've worked with rubber samples from belts, tires and gaskets in my SEM without any significant problems, despite the amount of volatile components present.

But these SEM were all oil pumped and a thin layer of oil was always present on the EDS detector and everywhere else too. I suspect amber might give you some volatility problems, so I'd consider using the smallest piece possible and if you had a low pressure SEM, I'd jump on that.

Too bad you not a member of the listserver, it's free and you could tell use how it works out for you.

Frank Karl

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Name: hadeel herzaallah

Organization: yarmouk university

Title-Subject: [Filtered] amber on sem

Message: hi ,can i study amber under the scanning electron microscope in high vacuum mode.

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Hello Bob,
I agree, Spurr's resin is great for viscosity but it's contrast is poor.
Also, I've never liked cutting Spurr's. In my hands it is brittle and
difficult to face and trim.
As a remedy, I now use a Spurr's / Epon combination. It give great contrast
and is MUCH easier to cut. Let me know if you would like my recipe and I can
forward it to you.
Take care,
Pat Kysar, C.E.M.T.
Chair, Certification Board, Microscopy Society of America
University of California, Davis
School of Medicine
Dept of Pathology and Laboratory Medicine
Electron Microscopy Lab

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Good Day: Â We are doing high pressure freezing and freeze substitution on
plant, animal and bacterial samples. We are using Epon resin for embedding
but struggle with its viscosity. Our experience with Spurr's has been
disappointing in that we get no or reverse contrast in the TEM. We love the
ease of use but can't make it work for imaging.
Â Â We are using recipes from Glauert (firm and the kit) and have both a
new kit and older stocks of the original ingredients. It doesn't seem to
make a difference. Â Â Â Does anyone have any advice on how we can make and
use this resin to produce samples that produce publication quality contrast
in our images? Thanks bob

--
Electron Microscopy Unit
New Science Complex
50 Stone Rd E
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Phone: Â 519-824-4120 Â ext. Â 56409
Fax Â : Â 519-837-1802

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Hello all

I am in the process of trying to get an used JSM-5400 for my EM lab near
Lisbon (non profit).
However the old (but functioning) microsope does not have the BS
detector (we need the best BSE images we can get) and the old NORAN
X-EDS attached to the microscope system does now work and can not be
recovered.
We are also lacking funding to get new detectors, so I would like to ask
if any of you have unused detectors that can be offered or obtained at a
low cost. We will take full care of the transport of course.
Commercial refurbished systems are also an option if not too expensive.
In case you wish to contact me directelly please use the e-mail
apamatos-at-kanguru.pt or apamatos-at-microventures.co

Many thanks
A.P. Alves de Matos
Biologist Ph.D.
Egas Moniz Electron Microscopy Unit, Caparica, Portugal

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Name: Valerie Lecomte

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Title-Subject: [Filtered] K250 carbon coating attachment

Message: Hi everyone,

Do you know where I can find a K250 Carbon Coating Attachment for an Emitech K550 sputter coater?

Thank you!

Valerie Lecomte

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From reasonable-meds12-at-idea-tele.com Tue Mar 25 01:41:54 2014
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Dear colleagues,
i have a question regarding the analysis of statistics, I apologize if this is not directly related to microscopy but I know that some of you are hardcore statisticians and I already got help in the past.
If I compare the effect of product A and B (whatever effect it doesn't matter) and I find with the Student-t test that p (A)=0,04 and p (B)=0,005, may I say that B works better than A?
I always thought that this interpretation of statistics is not allowed but I now see it in a recent Science paper and I surprised.
Many thanks in advance.
Stephane

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2, 38 -- From: Stephane Nizet {nizets2-at-yahoo.com}
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From auosueoa-at-yota.ru Tue Mar 25 06:41:23 2014
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Stephane,
At the risk of making a total fool of myself, I'll jump
in with the disclaimer that I am not a statistician.

But from what I understand, the first problem with what you
wrote is that p values represent a probalistic argument and are not
equalities. So should be p { x, never p = x. (This probably doesn't
matter to your story but just mentioning it)

Next, a t-test is typically done to tell whether two means
are equal. So in your case I assume that there were two experiments, one
for A and the other for B but in each case there was a control
treatment, "C". Thus, the first test (A vs C) shows that there is no
evidence to reject the hypothesis that the means are equal. The reported
difference would happen by chance more than once in twenty times, which
is generally considered not good enough to be "evidence". Though it can
be taken as a 'trend'. But the next test (B vs C) shows that the
equality of means can be rejected because the experimentally observed
difference should happen by chance less than once per hundred times.

So from this, there is no evidence that A works (ie,
differs from control). But there is evidence that B does. Therefore
seems only a logician (smile) would object to claiming that B works
better than A.

Anyway that is my take from the trenches. Maybe some
professionals will weigh in?

As ever,
Tobias

On 3/25/14, 5:54 AM, nizets2-at-yahoo.com wrote:
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} Dear colleagues,
} i have a question regarding the analysis of statistics, I apologize if this is not directly related to microscopy but I know that some of you are hardcore statisticians and I already got help in the past.
} If I compare the effect of product A and B (whatever effect it doesn't matter) and I find with the Student-t test that p (A)=0,04 and p (B)=0,005, may I say that B works better than A?
} I always thought that this interpretation of statistics is not allowed but I now see it in a recent Science paper and I surprised.
} Many thanks in advance.
} Stephane
}

--
__ ___ ^ ___ ___ Tobias I. Baskin
/ \ / / \ / \ Professor
/ / / / \ \ \ Biology Department
/ __/ /__ /___ \ \ \__ 611 N. Pleasant St.
/ / / \ \ \ University of Massachusetts
/ / / \ \ \ Amherst, MA, 01003 USA
/ /___ / \ \___/ \_____ 413 - 545 - 1533
www.bio.umass.edu/biology/baskin

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

Also not a statistician by trade, nor do I play one on TV. I do use them, though. A p value result in a statistical test tells you the probability of obtaining a result. Typically one sets an alpha value at 0.05 as a limit to exceed where you accept or reject the null hypothesis. Or, to simplify, in the case of a t-test for 2 means, your null hypothesis is "the 2 means are the same", and you set an alpha error limit at 1 out of 20, or 0.05. You perform the test and get a p value of 0.03, which is below alpha (YAY!), and report it as p {0.05 and conclude you can reject the null hypothesis and decide the 2 means are different. Statistically you are saying that if you did the experiment 20 times, you would expect this outcome 19 times. The p value tells you nothing about the magnitude of the difference between the means. It only tells you that one test is even more (or less) likely to have rejected the null hypothesis of "no difference". So, yes, the situation as you (Stephane) described would be an "over interpretation". You should be surprised to see such a conclusion in a Science paper. I would be shocked.

If you want to compare the magnitude of difference of two conditions (say, A and B) from a control (C), you would choose to run a statistical test built to tell you that, like a simple one-way analysis of variance (ANOVA). There are post-hoc tests to the ANOVA which allow you to compare the magnitude of the A to C and B to C differences, with yet another p value to report how confident you are about the size of that difference.

This brings me to mention a pet peeve, which is the over-use of Student's t-test in probing a set of data. When a t-test is run multiple times on the same set (or subset) of data, the alpha value (or probability of committing a Type I error of accepting a false positive) increases. This means that if you plan to do that, you need to set a lower alpha value, like 0.01, in order to find reliable effects. This is why more complex statistics like ANOVA are helpful ... you can still use 0.05 for alpha while simultaneously looking for differences between multiple combinations of data in a single test, then probe for trends and magnitudes with post-hoc tools. Just my $0.02 ...

Thomas Trusk, PhD
Director, Josh Spruill Imaging Facility
Dept of Regenerative Medicine and Cell Biology
Medical University of South Carolina
Charleston, SC 29425

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

Stephane,
At the risk of making a total fool of myself, I'll jump in with the disclaimer that I am not a statistician.

But from what I understand, the first problem with what you wrote is that p values represent a probalistic argument and are not equalities. So should be p { x, never p = x. (This probably doesn't matter to your story but just mentioning it)

Next, a t-test is typically done to tell whether two means are equal. So in your case I assume that there were two experiments, one for A and the other for B but in each case there was a control treatment, "C". Thus, the first test (A vs C) shows that there is no evidence to reject the hypothesis that the means are equal. The reported difference would happen by chance more than once in twenty times, which is generally considered not good enough to be "evidence". Though it can be taken as a 'trend'. But the next test (B vs C) shows that the equality of means can be rejected because the experimentally observed difference should happen by chance less than once per hundred times.

So from this, there is no evidence that A works (ie, differs from control). But there is evidence that B does. Therefore seems only a logician (smile) would object to claiming that B works better than A.

Anyway that is my take from the trenches. Maybe some professionals will weigh in?

As ever,
Tobias

}
}
} Dear colleagues,
} i have a question regarding the analysis of statistics, I apologize if this is not directly related to microscopy but I know that some of you are hardcore statisticians and I already got help in the past.
} If I compare the effect of product A and B (whatever effect it doesn't matter) and I find with the Student-t test that p (A)=0,04 and p (B)=0,005, may I say that B works better than A?
} I always thought that this interpretation of statistics is not allowed but I now see it in a recent Science paper and I surprised.
} Many thanks in advance.
} Stephane
}

--
__ ___ ^ ___ ___ Tobias I. Baskin
/ \ / / \ / \ Professor
/ / / / \ \ \ Biology Department
/ __/ /__ /___ \ \ \__ 611 N. Pleasant St.
/ / / \ \ \ University of Massachusetts
/ / / \ \ \ Amherst, MA, 01003 USA
/ /___ / \ \___/ \_____ 413 - 545 - 1533
www.bio.umass.edu/biology/baskin

==============================Original Headers==============================
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From drugstore_affordable9-at-swfastener.com Tue Mar 25 14:37:52 2014
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And Students t-test, ANOVA & others also assume data points are normally distributed,another wrinkle...

Dr Rosemary White
CSIRO Plant Industry
GPO Box 1600
Canberra, ACT 2601
Australia

T 61 2 6246 5475
F 61 2 6246 5334
________________________________________
X-from: truskt-at-musc.edu [truskt-at-musc.edu]
Sent: Wednesday, 26 March 2014 3:21 a.m.
To: White, Rosemary (PI, Black Mountain)

Also not a statistician by trade, nor do I play one on TV. I do use them, though. A p value result in a statistical test tells you the probability of obtaining a result. Typically one sets an alpha value at 0.05 as a limit to exceed where you accept or reject the null hypothesis. Or, to simplify, in the case of a t-test for 2 means, your null hypothesis is "the 2 means are the same", and you set an alpha error limit at 1 out of 20, or 0.05. You perform the test and get a p value of 0.03, which is below alpha (YAY!), and report it as p {0.05 and conclude you can reject the null hypothesis and decide the 2 means are different. Statistically you are saying that if you did the experiment 20 times, you would expect this outcome 19 times. The p value tells you nothing about the magnitude of the difference between the means. It only tells you that one test is even more (or less) likely to have rejected the null hypothesis of "no difference". So, yes, the situation as you (S!
tephane) described would be an "over interpretation". You should be surprised to see such a conclusion in a Science paper. I would be shocked.

If you want to compare the magnitude of difference of two conditions (say, A and B) from a control (C), you would choose to run a statistical test built to tell you that, like a simple one-way analysis of variance (ANOVA). There are post-hoc tests to the ANOVA which allow you to compare the magnitude of the A to C and B to C differences, with yet another p value to report how confident you are about the size of that difference.

This brings me to mention a pet peeve, which is the over-use of Student's t-test in probing a set of data. When a t-test is run multiple times on the same set (or subset) of data, the alpha value (or probability of committing a Type I error of accepting a false positive) increases. This means that if you plan to do that, you need to set a lower alpha value, like 0.01, in order to find reliable effects. This is why more complex statistics like ANOVA are helpful ... you can still use 0.05 for alpha while simultaneously looking for differences between multiple combinations of data in a single test, then probe for trends and magnitudes with post-hoc tools. Just my $0.02 ...

Thomas Trusk, PhD
Director, Josh Spruill Imaging Facility
Dept of Regenerative Medicine and Cell Biology
Medical University of South Carolina
Charleston, SC 29425

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

Stephane,
At the risk of making a total fool of myself, I'll jump in with the disclaimer that I am not a statistician.

But from what I understand, the first problem with what you wrote is that p values represent a probalistic argument and are not equalities. So should be p { x, never p = x. (This probably doesn't matter to your story but just mentioning it)

Next, a t-test is typically done to tell whether two means are equal. So in your case I assume that there were two experiments, one for A and the other for B but in each case there was a control treatment, "C". Thus, the first test (A vs C) shows that there is no evidence to reject the hypothesis that the means are equal. The reported difference would happen by chance more than once in twenty times, which is generally considered not good enough to be "evidence". Though it can be taken as a 'trend'. But the next test (B vs C) shows that the equality of means can be rejected because the experimentally observed difference should happen by chance less than once per hundred times.

So from this, there is no evidence that A works (ie, differs from control). But there is evidence that B does. Therefore seems only a logician (smile) would object to claiming that B works better than A.

Anyway that is my take from the trenches. Maybe some professionals will weigh in?

As ever,
Tobias

}
}
} Dear colleagues,
} i have a question regarding the analysis of statistics, I apologize if this is not directly related to microscopy but I know that some of you are hardcore statisticians and I already got help in the past.
} If I compare the effect of product A and B (whatever effect it doesn't matter) and I find with the Student-t test that p (A)=0,04 and p (B)=0,005, may I say that B works better than A?
} I always thought that this interpretation of statistics is not allowed but I now see it in a recent Science paper and I surprised.
} Many thanks in advance.
} Stephane
}

--
__ ___ ^ ___ ___ Tobias I. Baskin
/ \ / / \ / \ Professor
/ / / / \ \ \ Biology Department
/ __/ /__ /___ \ \ \__ 611 N. Pleasant St.
/ / / \ \ \ University of Massachusetts
/ / / \ \ \ Amherst, MA, 01003 USA
/ /___ / \ \___/ \_____ 413 - 545 - 1533
www.bio.umass.edu/biology/baskin

==============================Original Headers==============================
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Could someone please let me know if one can glow-discharge grids using a JEOL JEE 4B/4C vacuum evaporator? If so, how would one go about doing such? The instruction manual does not cover this. (These would be copper grids filmed with formvar-carbon to be used for negative staining purposes.)

Thank you,

Jaci

Jaclynn Lett
Senior Research Technician, EM Facility
Research Center for Auditory and Vestibular Studies
Department of Otolaryngology
Washington University School of Medicine
660 S. Euclid Ave., Campus Box 8115
St. Louis, MO 63110
Office: 314-747-7257
Fax: 314-747-7230
Email: lettj-at-ent.wustl.edu
http://otocore.wustl.edu

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Title-Subject: [Filtered] LaB6 filament life time

Message: Hi everyone,
Is there anyone using a LaB6 filament on their SEM...
I would like to know what is the filament life time? Is it a lot shorter
if we use the filament on the second saturation point?

I currently using a LaB6 filament on a Zeiss EVO and I have to put the
Filament Intensity higher and higher everyday to reach the second
saturation peak. Do you think my filament is dying or it is another
problem? It has 1800 hours lifetime.

Thank you for your help!

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Hi Valerie,
I am using LaB6 cathodes since nearly ten years in my Philips / FEI 525 SEM.
1800 hours of beamtime with one filament sounds good. That`s what I get out also, if the vacuum is perfect.

Remember: the cristal tip of the cathode gets sputtered away with time, so near the end of the lifetime you need to adjust the
distance tip to wehnelt aperture closer to get the same current.
Or you increase (and this is not the best way) the heating current (which you might need to do anyway with this needle-like
cristal structure remaining...).
Remember also that LaB6 deposits on the wehnelt aperture, leaving an insulating layer you have to clean away.

I would go having a look at your cathode-wehnelt assembly and adjust it new. Then you might be lucky to get some 100 hours of
lifetime. Depends how the cathode looks like...

Best wishes,
Stefan

-----------------------------------------------------
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D - 97072 Wuerzburg Germany
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++49-931-7848701 Fax
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Am 27.03.14 02:24, schrieb zaluzec-at-microscopy.com:
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}
} Title-Subject: [Filtered] LaB6 filament life time
}
} Message: Hi everyone,
} Is there anyone using a LaB6 filament on their SEM...
} I would like to know what is the filament life time? Is it a lot shorter
} if we use the filament on the second saturation point?
}
} I currently using a LaB6 filament on a Zeiss EVO and I have to put the
} Filament Intensity higher and higher everyday to reach the second
} saturation peak. Do you think my filament is dying or it is another
} problem? It has 1800 hours lifetime.
}
} Thank you for your help!
}
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==============================Original Headers==============================
10, 22 -- From stefan.diller-at-t-online.de Thu Mar 27 03:06:53 2014
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From express-meds10-at-lindemulder.org Thu Mar 27 05:19:29 2014
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Hi Stefan, et al.,

Kimball Physics has some useful information in their technical notes on
the operation of LaB6 cathodes. You can find it on the Kimball Physics
web site under Cathodes/Emitters and then Technical information.

The major reason for LaB6 end-of-life is due to loss of brightness of
the emitter. Most current emitters are [100] oriented single crystals
with a 90 degree cone angle and a roughly 15um flat at the tip. As the
tip is used, LaB6 evaporates from it and the tip recedes. There is a
faster recession on the [110] faces and as the tip recedes, there is a
gradual loss of the flat, i.e. the tip gradually sharpens. Since the
emitting area drops, the total current does also. It is also likely
that since the cathodes are heated from the edges by the graphite
support that the tip is now running cooler that previously.

As an operating note, overheating LaB6 cathodes significantly reduces
lifetime. Don't oversaturate! Taking the temperature from the nominal
1800K to 1850K increases the evaporation rate by 3X. This means that the
lifetime will go from ~2000hrs to less than 700!

You will see deposits from the cathode on the inside of the wehnelt.
These should be a purplish color, indicative of LaB6. If the deposits
are white, you have formed Lanthanum oxide. Oxide formation means that
the gun vacuum is not adequate. Also, the oxide is an insulator whereas
the boride is a semiconductor. This means that as the oxide builds up,
you will get charging effects on the inside of the wehnelt. This will
eventually result in a flickering beam intensity.

Cheers,
Henk

On 3/27/2014 4:08 AM, stefan.diller-at-t-online.de wrote:
}
}
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
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} ----------------------------------------------------------------------------
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} Hi Valerie,
} I am using LaB6 cathodes since nearly ten years in my Philips / FEI 525 SEM.
} 1800 hours of beamtime with one filament sounds good. That`s what I get out also, if the vacuum is perfect.
}
} Remember: the cristal tip of the cathode gets sputtered away with time, so near the end of the lifetime you need to adjust the
} distance tip to wehnelt aperture closer to get the same current.
} Or you increase (and this is not the best way) the heating current (which you might need to do anyway with this needle-like
} cristal structure remaining...).
} Remember also that LaB6 deposits on the wehnelt aperture, leaving an insulating layer you have to clean away.
}
} I would go having a look at your cathode-wehnelt assembly and adjust it new. Then you might be lucky to get some 100 hours of
} lifetime. Depends how the cathode looks like...
}
}
} Best wishes,
} Stefan
}
}
}
} -----------------------------------------------------
} Stefan Diller - Scientific Photography
} Arndtstrasse 22
} D - 97072 Wuerzburg Germany
} ++49-931-7848700 Phone
} ++49-931-7848701 Fax
} ++49-175-7177051 Mobile
}
} Websites:
} www.nanoflight.info
} www.stefan-diller.com
} www.electronmicroscopy.info
} www.elektronenmikroskopie.info
} www.zwillingsprojekt.de
} www.assisi.de
} Anfahrt: http://Mail.map24.com/Stefan.Diller
} -----------------------------------------------------
}
} Am 27.03.14 02:24, schrieb zaluzec-at-microscopy.com:
} } ----------------------------------------------------------------------------
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} } Organization: IOS services geo/Sherbrooke University
} }
} } Title-Subject: [Filtered] LaB6 filament life time
} }
} } Message: Hi everyone,
} } Is there anyone using a LaB6 filament on their SEM...
} } I would like to know what is the filament life time? Is it a lot shorter
} } if we use the filament on the second saturation point?
} }
} } I currently using a LaB6 filament on a Zeiss EVO and I have to put the
} } Filament Intensity higher and higher everyday to reach the second
} } saturation peak. Do you think my filament is dying or it is another
} } problem? It has 1800 hours lifetime.
} }
} } Thank you for your help!
} }
} } Login Host: 24.37.116.198
} } Listserver Email Form V - 20120416
} } ---------------------------------------------------------------------------
} }
} }
}
} ==============================Original Headers==============================
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--

Hendrik O. Colijn

*C*enter for *E*lectron *M*icroscopy & *A*nalysi*S*

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cemas.osu.edu

"Time is that quality of nature which keeps things from happening all at
once. Lately it doesn't seem to be working."

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Hi Valerie

I think that you are doing very well to keep your filament for 1800 hours.
Please be aware that the manufacturers in the past have suggest LaB6 cathode
cleaning at least every 500 hours! A cleaner cathode will help the vacuum
level in the gun and prolong life.

Be aware that the level of heating with any type of filament will be a big
factor in filament life. Are you sure that you must use the settings you
mention, could you lower the heating level? But I say again, you have had a
pretty good filament life and you are right, you now must consider a
replacement?

Good luck

Steve

Steve Chapman FRMS
Protrain for Consultancy and Courses in Electron Microscopy
Mobile +44 (0)7711 606967 Fax +44 (0)1280 814007
www.emcourses.com

Email: valerie.lecomte-at-usherbrooke.ca
Name: Valerie Lecomte

Organization: IOS services geo/Sherbrooke University

Title-Subject: [Filtered] LaB6 filament life time

Message: Hi everyone,
Is there anyone using a LaB6 filament on their SEM...
I would like to know what is the filament life time? Is it a lot shorter if
we use the filament on the second saturation point?

I currently using a LaB6 filament on a Zeiss EVO and I have to put the
Filament Intensity higher and higher everyday to reach the second saturation
peak. Do you think my filament is dying or it is another problem? It has
1800 hours lifetime.

Thank you for your help!

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Postdoctoral fellowship opportunity in focused ion beam nanotomography
at the US Naval Research Laboratory in Washington, DC.

The postdoctoral position is jointly funded by the Office of Naval
Research and the NASA Origins program. The focus of the position will be
on development of 3D imaging and tomographic reconstruction of materials
down to the 10-nm scale, using FIB-SEM slice-and-view methods.
Demonstrated skill in the operation of an SEM is required. FIB-SEM and
image processing experience, including experience with the Avizo Fire
software package, are desirable. The project will involve analysis of
meteorites and materials of more direct Navy interest, such as
barnacles, or synthetic nanocomposite materials. The salary for this
position is set at the NRL/NRC/ASEE fellow rate of $74,872. Interested
candidates may send inquiries to rhonda.stroud at nrl.navy.mil. Please
include a CV and publication list.

Due to security regulations at NRL, this position requires US
citizenship or permanent residency.

Rhonda Stroud

--

___________________________________
Head, Nanoscale Materials Section
Naval Research Laboratory
4555 Overlook Ave SW
Washington, DC 20375
(V) 202-404-4143 (F) 202-767-1697

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A postdoctoral fellowship position is available now at the Naval
Research Laboratory in Washington, DC, for the study of the mineralogy
and petrology of meteorites and returned samples. The successful
candidate will use STEM-based spectroscopy for mineralogical studies,
including oxidation state analysis. The postdoc will coordinate the STEM
studies with synchrotron-based studies conducted by team members at the
partnering NASA SSERVI RIS^4 E institutions, for the analysis of analog
materials, meteorites and returned lunar, Stardust and/or Hayabusa
samples. Expertise in transmission electron microscopy is required, and
familiarity with EELS and advanced sample preparation methods is
preferred. Instrumentation at NRL for these studies includes a Nion
UltraSTEM 200 (to be installed June 2014), and a JEOL 2200FS. The salary
for this position is set at the NRL/NRC/ASEE fellow rate of $74,872.
Interested candidates may send inquiries to rhonda.stroud at
nrl.navy.mil {mailto:rhonda.stroud-at-nrl.navy.mil} . Please include a CV
and publication list.

Due to security regulations at NRL, this position requires US
citizenship or permanent residency.

Rhonda Stroud

--

___________________________________
Head, Nanoscale Materials Section
Naval Research Laboratory
4555 Overlook Ave SW
Washington, DC 20375
(V) 202-404-4143 (F) 202-767-1697

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Regarding:

" If I compare the effect of product A and B (whatever effect it doesn't
matter) and I find with the Student-t test that p (A)=0,04 and p (B)=0,005,
may I say that B works better than A?"

Disclaimer - I'm not a statistician, just a heavy user of stats.

My long-winded way of saying No:

The Student t-test [1,2] probably used is better described as a test of the
means of two normal distributions with variances unknown, but equal [3]. It
assumes data are univariate, continuous, and normal distributed, and data
are collected randomly [3, 4]. It should not be used when data are ranked,
not approximately normally distributed, and more than two groups are to be
compared [4]. It is not valid for multiple comparisons (because of additive
errors) or where group sizes are very unequal [4]. The test is robust for
moderate departures from normality, and when N1 and N2 are approximately
equal it is robust from departures of homogeneity (equality) of variances
[3,4]. An F-test is commonly performed to check the homogeneity of the data
(determine if the variances are equal). If they are homogeneous, then for
comparing the Mean of two groups, the t-test can be used. If they are
heterogeneous, then a modified t-test (Cochran t-test; test of the means of
two normal distributions with variances unknown, but unequal) [3-5] can be
used. If the data are not normally distributed, then a non-parametric
Student's test could be performed [6]. In addition, "although one might
presume that a small P value indicates the presence of strong evidence
against the null, such is not necessarily the case" [7], e.g., a better
p-value doesn't mean better test method/group. Since it is a test of the
Means (is this data set different from another), it says very little about
the rest of the data spread, which may be more important.

If one wishes to compare more than two groups (Control, A, and B), then
ANOVA is usually performed. The typical use of ANOVA is to demonstrate both
a difference and through post-hoc analysis a significance for treatment sets
(Class I and II uses according to Eisenhart [8]) [4]. The use of ANOVA is
predicated on the assumption of normality [8, 9], with varying consequences
in both Class I and Class II uses [9]. A common example of a non-normal
data set would be counting, such fiber or mineral grain or blood cell
counting as these better following a binomial-based distribution - poisson
or negative binomial [10-19]. In these cases, the count data should be
transformed by taking the square root to "normalize" the data [20-22] prior
to performing ANOVA. Even if ANOVA is used, the strength of the effect
should still be looked at carefully/cautiously [23-25]. Furthermore, a
different multi-comparison method may be more appropriate [26-30].

1. Student: Probable Error of a Correlation Coefficient, Biometrika, 6,
2-3, 302-310, 1908. (pen name for Gosset)
2. Student: The Probable Error of a Mean, Biometrika, 6, 1, 1-25, 1908.
3. Hines, Wulliam H., and Douglas C. Montgomery: Probability and
Statistics in Engineering and Management Science, 3rd ed. John Wiley &
Sons, NY. NY. 1990.
4. Gad, Shayne, and Carrol S. Weil: Statistics and Experimental Design
for Toxicologists, 2nd Ed. CRC Press, Boca Raton, FL. 1991.
5. Cochran, William G., Gertrude M. Cox: Experimental Designs, 2nd ed.
John Wiley & Sons, NY, NY. 1975.
6. Walsh, John E.: Some Nonparametric Tests for Student's Hypothesis
in Experimental Designs, J Am Stat Assoc, 47, 259, 401-415, 1952.
7. Berger, James, and Thomas Sellke Testing a Point Null Hypothesis,
The Irreconcilability of P Values and Evidence, J Am Stat Assoc, 82, 397,
112-122, 1987.
8. Eisenhart, The Assumptions Underlying the Analysis of Variance,
Biometrics, 3, 1, 1-21, 1947.
9. Cochran, Some Consequences When the Assumptions for the Analysis of
Variance are not Satisfied, Biometrics, 3, 1, 22-38), 1947.
10. NIOSH: Proficiency Analytical Testing Program & Environmental Lead
Proficiency Analytical Testing Program (PAT & ELPAT) Programs. NIOSH Pub
95-104. USDHHS, Cincinnati, OH. November, 1994.
11. Schlect, P.C., and S.A. Shulman: Performance of asbestos fiber
counting laboratories in the NIOSH proficiency analytical testing (PAT)
program. AIHAJ 47(5):259-269. 1986.
12. Reist, A Further Note on the Reliability of Membrane Filter Dust
Sample Evaluation by Microscope Counting, Ann Occ Hyg, 13, 4, 201-204, 1970.
13. Hoyes, A Routine for the Control of Perform of Microscopists Eval
Airborne Asbestos Fibre Samples on Membrane Filters by PCM, Ann Occ Hyg, 23,
4, 381-401, 1980
14. Sniegowski, A Note on Reliability of Membrane Filter Dust Sample
Evaluation by Microscope Counting, Ann Occ Hyg, 9, 2, 65-67, 1966.
15. Chapman, Harlow M., and Russell C. Ruhf: Dust Counting Reliability,
AIHA Qtr, 16, 3, 210-209, 1955.
16. Reid, George W.: Calculation of Counting Statistics, AIHA Qtr, 13,
1, 29-32, 1952.
17. Oehlert, Statistical Anal of Asbestos Fibre Counts, Environmetrics,
6, 115-126, 1995.
18. Chayes, Felix: Petrographic Modal Analysis. John Wiley & Sons, NY.
NY. 1956.
19. Student: On the Error of Counting with a Haemacytometer,
Biometrika, 5, 3, 351-360, 1907.
20. Barlett, M.S.: The Square Root Transformation in Analysis of
Variance, Suppl to J Roy Stat Soc, 3, 1, 68-78, 1936.
21. Bartlett, M.S.: The Use of Transformations, Biometrics, 3, 1,
39-52, 1947.
22. Havics, A: Asbestos Fiber Counting by Different optical techniques,
Microscope, 58, 2, 51-62, 2010.
23. Ronis, David L.: Comparing the Magnitude of Effects in Anova
Designs, Ed Psychol Meas, 41, 4, 993-1000, 1981.
24. Vaughan, Graham, and Michael C. Corbalis: Beyond Tests of
Significance, Est Strength of Effects in Slected ANOVA Design, Psychol Bull,
72, 3, 204-213, 1969.
25. Camilli, Gregory, and Lorrie A. Shepard: The inadequacy of ANOVA
for detecting test bias, J Ed Stats, 12, 1, 87099, 1987.
26. Riegelman, Richard K.: Studying a Study and Testing a Test, 5th ed.
Lippincott Williams & Wilkins. 2005.
27. Shaffer, Juliet Popper: Multiple hypothesis testing, Ann Rev
Psychol, 46, 561-584, 1995
28. Holland, Burt, and Margaret DiPonzio Copenhaver: Improved
Bonferroni-Type Multiple Testing Procedures, Physcol Bull, 104, 1, 145-149,
1988
29. Einot, Israeil, and K.R. Gabrie: A study of the powers of several
methods of multiple comparisons, J Am Stat Assoc, 70, 351, 574-583, 1975.
30. Curran-Everett, Douglas: Multiple comparisons, philosophies and
illustrations, Am J Physiol Reg Integ Comp Physiol, 279, R1-R-8, 2000.

Tony
.........................................
Andrew Anthony "Tony" Havics, CIH, PE
Environmental, Health & Safety, Microscopy, Materials Science & Forensic
Engineering
pH2, LLC
5250 E US Highway 36, Suite 830
Avon, IN 46123
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-----Original Message-----
X-from: nizets2-at-yahoo.com [mailto:nizets2-at-yahoo.com]
Sent: Tuesday, March 25, 2014 5:59 AM
To: ph2-at-sprynet.com

Dear colleagues,
i have a question regarding the analysis of statistics, I apologize if this
is not directly related to microscopy but I know that some of you are
hardcore statisticians and I already got help in the past.
If I compare the effect of product A and B (whatever effect it doesn't
matter) and I find with the Student-t test that p (A)=0,04 and p (B)=0,005,
may I say that B works better than A?
I always thought that this interpretation of statistics is not allowed but I
now see it in a recent Science paper and I surprised.
Many thanks in advance.
Stephane

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Contents Retrieved from Microscopy Listserver Archives
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A recent reply to this thread was rather technical and dense. In
contrast "Naked Statistics" by Charles Wheelan, which I have just
finished reading, is an easy read and an accessible guide to the power
and traps of doing statistical analysis.

It might be a good place to gain perspective.

Alwyn Eades
Department of Materials Science and Engineering
Lehigh University

==============================Original Headers==============================
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Contents Retrieved from Microscopy Listserver Archives
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I find "Intuitive Biostatistics" by Harvey Motulsky as quite accessible for the statistical novice. He is the guy behind Graphpad statistical software.

Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
phillipst-at-missouri.edu

http://biology.missouri.edu/people/?person=88
http://www.biotech.missouri.edu/mcc/

-----Original Message-----
X-from: jae5-at-lehigh.edu [mailto:jae5-at-lehigh.edu]
Sent: Thursday, March 27, 2014 2:53 PM
To: Phillips, Thomas E.

A recent reply to this thread was rather technical and dense. In contrast "Naked Statistics" by Charles Wheelan, which I have just finished reading, is an easy read and an accessible guide to the power and traps of doing statistical analysis.

It might be a good place to gain perspective.

Alwyn Eades
Department of Materials Science and Engineering Lehigh University

==============================Original Headers==============================
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Dear Listers,

Just a reminder about next Wednesday's webinar: Brightfield Color
imaging: Is what you saw what you get?
Wednesday, April 2, 2014, 1:00PM EDT
To
register:
{http://scientific.datacolor.com/resources/events/} http://scientific.datacolor.com/resources/events/

Guest Speaker: Dr. Eduardo Rosa-Molinar
Dr. Rosa-Molinar is a Professor in the Department of Biology at the
University of Puerto Rico Rio Piedras. He serves on the FASEB Board
of Directors, and has served in numerous positions with The
Histochemical Society, the Society for Integrative and Comparative
Biology, the Society for Neuroscience, and is currently the Chair of
the Andor/Bitplane Advanced Imaging Centers.

Interpretation and communication are the essence of science. But
what happens when what you see in the microscope is not what you see
on your monitor? Or your laptop? Or that your colleague sees when
he opens that all-important image you sent him? Ironically, these
variations in the image are often most pronounced in that good old
workhorse, brightfield imaging of stained samples. This webinar will
review best practices in brightfield imaging then discuss how
Datacolor's ChromaCal microscope and monitor systems can help you to
standardize those images so that what you saw in the microscope is
what you get on your screen.

==============================Original Headers==============================
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From drugstore-express6-at-corbina.ru Fri Mar 28 02:08:55 2014
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After seeing these prestigious references, I'm guess I shouldn't bother mentioning "Statistics for Dummies" by Rumsey.

Opps! Too late..........

Stay safe
Frank

-----Original Message-----
X-from: jae5-at-lehigh.edu [mailto:jae5-at-lehigh.edu]
Sent: Thursday, March 27, 2014 4:01 PM
To: Frank Karl

A recent reply to this thread was rather technical and dense. In
contrast "Naked Statistics" by Charles Wheelan, which I have just
finished reading, is an easy read and an accessible guide to the power
and traps of doing statistical analysis.

It might be a good place to gain perspective.

Alwyn Eades
Department of Materials Science and Engineering
Lehigh University

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The Northern California Society for Microscopy Spring Meeting, featuring presentations by Kent McDonald and Ryan McGorty, will be held Thursday, April 17, 2014 6:00 PM to 9:00 PM.

Please RSVP to info-at-ncsmicroscopy.org

Scotts of Palo Alto
#1 Town and Country Village
Palo Alto, California
http://scottsseafoodpa.com/

Featured Speakers Are:

Kent McDonald, PhD, Electron Microscopy Lab, UC Berkeley

Rapid Specimen Processing Methods for Cryofixed Samples

New methods for freeze substitution have been developed that
require only basic laboratory tools. Excellent freeze substitution
results can be obtained in as little as 90min for cells of small
volume. Rapid epoxy resin infiltration can be achieved by centrifugation
followed by polymerization at 100 C for 1.5–2 h. Total processing time
from freezing to blocks ready to section is about 6 h. Comparative
results will be presented.

Ryan McGorty, PhD, UC San Francisco, School of Pharmacy

Three-dimensional super-resolution imaging of tissue samples

Single-molecule localization microscopy methods are often used to
study samples at or very near the coverslip surface with a spatial
resolution of around 20nm. However, a number of challenges emerge when
trying to image even a few microns past the coverslip surface,
particularly when trying to accurately localize molecules in 3D. After
first explaining the principle of the super-resolution microscopy
technique we use, I will discuss multiple methods we have recently
developed that allow us to image deeper into tissue samples.

No Cost to Current Members. $25 for Non-Members.

Bring your check, payable to NCSM, for 2014 dues and enjoy a 3-course meal and great speakers.
Annual dues for regular members are $40; students, $20; and a
corporate membership (which includes one person) is $100, payable to "NCSM"

_______________________
Roseann Csencsits, PhD
Scientist in Charge, Downing TEM Tools
Lawrence Berkeley Lab 01-365
1 Cyclotron Road
Berkeley, CA 94720
510-486-4548

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I was all excited when I heard someone on this campus is going to get a
FIB/SEM, but I never imagined this...

https://www.youtube.com/watch?feature=player_embedded&v=IV2_da-Km0s

Aloha,
Tina

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

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Interesting.
But why an operator was in latex gloves?
Vladimir

Vladimir M. Dusevich, Ph.D.
Electron Microscope Lab Director
(816) 235-2072
371 School of Dentistry
650 E. 25th Street
Kansas City, MO 64108-2784

-----Original Message-----
X-from: tina-at-pbrc.hawaii.edu [mailto:tina-at-pbrc.hawaii.edu]
Sent: Friday, March 28, 2014 1:29 PM
To: Dusevich, Vladimir

I was all excited when I heard someone on this campus is going to get a FIB/SEM, but I never imagined this...

https://www.youtube.com/watch?feature=player_embedded&v=IV2_da-Km0s

Aloha,
Tina

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

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12, 30 -- Fri, 28 Mar 2014 15:44:52 -0500
12, 30 -- From: "Dusevich, Vladimir" {DusevichV-at-umkc.edu}
12, 30 -- To: "Microscopy-at-microscopy.com" {Microscopy-at-microscopy.com}
12, 30 -- Subject: RE: [Microscopy] Etching a castle onto a grain of sand with FIB/SEM
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From drugs_canadian3-at-tktelekom.pl Sat Mar 29 07:15:38 2014
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Email: modla-at-dbi.udel.edu
Name: Shannon Modla

Organization: University of Delware

Title-Subject: [Filtered] Free Ditabis Imaging Plate Scanner

Message: Hi Everyone!

We have a Ditabis Imaging Plate Scanner that we are giving away. The system comes with the lightbox
eraser, software CDs, cables, 30 imaging plates, and the cassettes that hold the plates. We also
have a PC with the software already installed.

The last time the system was used, it was in good working condition. Since we've upgraded to a new
TEM with a CCD camera (it doesn't even have the option for film now), we have not needed to use the
Ditabis system.

For those not familiar with the Ditabis, it consists of special imaging plates that you load into
the TEM same as film. After you expose the beam to the film, it can be removed and scanned using
the Ditabis Micron scanner, and then the high resolution image is available in digital format. The
film can then be erased on the lightbox and repeatedly reused.

For anyone interested, please contact me. If you want the system, you will either need to come pick
it up or be responsible for the shipping/handling. The scanner itself probably weights a couple
hundred pounds. We are located at the University of Delaware in Newark, DE.

Shannon Modla

Research Associate II
University of Delaware
Delaware Biotechnology Institute
BioImaging Center
15 Innovation Way, Suite 117
Newark, DE 19711
modla-at-dbi.udel.edu

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18, 28 -- From microscopylistserver-noreply-at-microscopy.com Sat Mar 29 08:01:50 2014
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From drugstore-cheapest12-at-suratifarsan.com Sun Mar 30 22:56:54 2014
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I'm trying to diagnose a vacuum problem with the Leica S430 that was
donated to the Pumping Station: One hackerspace in Chicago. We are, to
our knowledge, the only hackerspace in the US with a working SEM, and we
are developing programming around making it accessible to the general
public.

When the scope arrived, it couldn't achieve vacuum better than 6e-5
torr. Over time, it did substantially better, and, after I changed the
roughing pump oil, it could get down, at best, to 1.8e-5 torr. Still,
factory specs say it should go to 2e-6, and I'd be much happier if it
were in the e-6 range.

However, I'd noticed strange problems where, after a week of running,
I'd check it and find that it seemed stuck at a rather awful vacuum of
1.2 to 1.4e-4 torr. The act of venting the chamber and pumping it down
again seemed to cure it, and it would pump down rather quickly to 3e-5
torr and slowly get better from there. The oil change didn't solve this.
I tried removing the oil mist / odor filter while the vacuum was stalled
in the 1.?e-4 torr range to see if it was creating back pressure, but
that didn't do much. Since then, I've replaced the oil mist and odor
filter elements. This helps somewhat, but not entirely, at removing the
hot oil reek when pumping down the chamber from atmospheric pressure.

Things have become substantially worse. Now, pumping down from
atmospheric pressure works as expected, but within hours, pressure rises
to the 1.2 to 1.4e-4 torr level again. Venting and pumping fixes it
temporarily until this happens again in a few hours. Thoughts:

Virtual leak / sample outgassing: I'm doubting this. I keep the scope at
vacuum 24x7 and haven't changed out the samples in months. (I'm
primarily teaching people to use the scope by looking at samples already
prepared. I don't have curriculum together on sample prep, although we
do have a sputter coater and critical point dryer.)

Physical leak: That might explain why I'm not getting below 1.8e-5, but
I don't see it explaining why the pressure rises so much after a
successful pump down. I've also wiped down the chamber and gun O-rings
recently with no effect.

Backing pump oil mist / odor filter back pressure: Possible, as I
rebuilt the filter recently and could have caused a problem, but this
didn't start immediately after I rebuild the filter, and when I tried
removing the filter before, it never helped.

Backing pump oil age: I changed it a little over 4 months ago. I suppose
this is possible, but I'd not had this problem with this frequency when
I was running oil that had been in the pump for a very long time from
the former owner.

Air filter dessicant: it's starting to change color in places. I know I
should heat it to drive out the moisture. But even if moist air is
coming into the chamber when I vent it, I'd think that would slow my
pump down time, not cause a normal pump down and reappear as a problem
later.

I wouldn't know if this were a problem with the Penning gauge. Usually,
after the Penning turns on, I see some sinusoidal oscillation in
pressure during initial pump down, but then it damps out and behaves
predictably otherwise.

So I'm left thinking this is either a turbomolecular pump problem, or a
roughing pump problem. The turbopump does make a soft high pitched
whine, which sometimes changes pitch, but I've heard that is normal. The
roughing pump does change its sound periodically. Especially when
pumping down, it sounds like it is running rough. The scope didn't have
a maintenance contract at its former home, so I question how much
preventative maintenance was done. If I had to guess, I'd question if
the scope needs a rebuild. I've seen kits available. (Edwards E2M12 I
think.)

We can't exactly afford thousands of dollars to bring in a SEM tech.
Buying a rebuild kit for the roughing pump would be financially doable,
but I've heard this is a lot of work (Have any of you rebuilt a
multi-stage rotary vane pump? How difficult?), and I'd probably need an
offsite area to do the rebuild because our "microscopy lab" is in a
corner of a dusty workshop. (Yes, I know this isn't ideal for a SEM....)
If possible, I'd really like more of an indication that the roughing
pump is the problem before trying this.

Have any of you seen problems like this? Any thoughts on how to proceed?

Thanks,
Ryan

==============================Original Headers==============================
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From discounted-meds2-at-eccreativesolutions.com Tue Apr 1 05:19:30 2014
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Announcing a 2014 REU in Materials Science and Engineering "Physical Properties of Materials" to be hosted at Mississippi State University

Don't miss this exciting opportunity for undergraduate research experience.

Details and information on how to apply can be found at www.abe.msstate.edu/reu/

Thanks,
Amanda Lawrence
Institute for Imaging and Analytical Technologies
Mississippi State University

==============================Original Headers==============================
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Dear Listers,

Just a reminder:
Wednesday, April 2, 2014, 1:00PM EDT
To
register:
{http://scientific.datacolor.com/resources/events/} http://scientific.datacolor.com/resources/events/

Interpretation and communication are the essence
of science. But what happens when what you see
in the microscope is not what you see on your
monitor? Or your laptop? Or that your colleague
sees when he opens that all-important image you
sent him? Ironically, these variations in the
image are often most pronounced in that good old
workhorse, brightfield imaging of stained
samples. This webinar will review best practices
in brightfield imaging then discuss how
Datacolor’s ChromaCal microscope and monitor
systems can help you to standardize those images
so that what you saw in the microscope is what you get on your screen.

Guest Speaker: Dr. Eduardo Rosa-Molinar
Dr. Rosa-Molinar is a Professor in the Department
of Biology at the University of Puerto Rico – Rio
Piedras. He serves on the FASEB Board of
Directors, and has served in numerous positions
with The Histochemical Society, the Society for
Integrative and Comparative Biology, the Society
for Neuroscience, and is currently the Chair of
the Andor/Bitplane Advanced Imaging Centers.

==============================Original Headers==============================
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6, 38 -- Subject: Webinar Reminder: Maintaining Color Integrity, with Dr.
6, 38 -- Eduardo Rosa-Molinar
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Email: see.wee.chee-at-gmail.com
Name: See Wee Chee

Title-Subject: [Filtered] Olympus TEM Cameras

Message: Hello everybody,

The side-mounted camera on our TEM has failed and we are looking for a replacement. We discovered
that Olympus is now in market with three side-mounted cameras. The specifications look pretty good
but we don't know of anyone that has one in service.

Has anyone here used an Olympus camera on their TEM before? Any feedback on performance, software
and support will be greatly appreciated. We are interested in the MegaView G2 and Veleta lines. Thanks!

See Wee

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Ryan,
I doubt that your Rotary pump (RP) is the problem. I'm not familiar with
the S430 but have some familiarity with the 1430 and 1430VP. First, check
and see if the turbopump is going into a "standby mode" at a reduced RPM.
In conjunctin with a leak, this could cause the vacuum deterioration that
you are seeing. More likely, the Penning gauge needs to be cleaned.
Determine the model and you should be able to find a user's manual on line
with instructions on how to clean it.

I have seen systems behave in a somewhat similar manner with oil diffusion
pumps due to either a lack of DP oil or because of issues in systems that
change the power settings to the DP in different pumping stages. This is
not generally the case with TMPs, so clean the Penning Gauge and see what
happens.

Ken Converse
owner

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

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To: kenconverse-at-qualityimages.biz

I'm trying to diagnose a vacuum problem with the Leica S430 that was
donated to the Pumping Station: One hackerspace in Chicago. We are, to
our knowledge, the only hackerspace in the US with a working SEM, and we
are developing programming around making it accessible to the general
public.

When the scope arrived, it couldn't achieve vacuum better than 6e-5
torr. Over time, it did substantially better, and, after I changed the
roughing pump oil, it could get down, at best, to 1.8e-5 torr. Still,
factory specs say it should go to 2e-6, and I'd be much happier if it
were in the e-6 range.

However, I'd noticed strange problems where, after a week of running,
I'd check it and find that it seemed stuck at a rather awful vacuum of
1.2 to 1.4e-4 torr. The act of venting the chamber and pumping it down
again seemed to cure it, and it would pump down rather quickly to 3e-5
torr and slowly get better from there. The oil change didn't solve this.
I tried removing the oil mist / odor filter while the vacuum was stalled
in the 1.?e-4 torr range to see if it was creating back pressure, but
that didn't do much. Since then, I've replaced the oil mist and odor
filter elements. This helps somewhat, but not entirely, at removing the
hot oil reek when pumping down the chamber from atmospheric pressure.

Things have become substantially worse. Now, pumping down from
atmospheric pressure works as expected, but within hours, pressure rises
to the 1.2 to 1.4e-4 torr level again. Venting and pumping fixes it
temporarily until this happens again in a few hours. Thoughts:

Virtual leak / sample outgassing: I'm doubting this. I keep the scope at
vacuum 24x7 and haven't changed out the samples in months. (I'm
primarily teaching people to use the scope by looking at samples already
prepared. I don't have curriculum together on sample prep, although we
do have a sputter coater and critical point dryer.)

Physical leak: That might explain why I'm not getting below 1.8e-5, but
I don't see it explaining why the pressure rises so much after a
successful pump down. I've also wiped down the chamber and gun O-rings
recently with no effect.

Backing pump oil mist / odor filter back pressure: Possible, as I
rebuilt the filter recently and could have caused a problem, but this
didn't start immediately after I rebuild the filter, and when I tried
removing the filter before, it never helped.

Backing pump oil age: I changed it a little over 4 months ago. I suppose
this is possible, but I'd not had this problem with this frequency when
I was running oil that had been in the pump for a very long time from
the former owner.

Air filter dessicant: it's starting to change color in places. I know I
should heat it to drive out the moisture. But even if moist air is
coming into the chamber when I vent it, I'd think that would slow my
pump down time, not cause a normal pump down and reappear as a problem
later.

I wouldn't know if this were a problem with the Penning gauge. Usually,
after the Penning turns on, I see some sinusoidal oscillation in
pressure during initial pump down, but then it damps out and behaves
predictably otherwise.

So I'm left thinking this is either a turbomolecular pump problem, or a
roughing pump problem. The turbopump does make a soft high pitched
whine, which sometimes changes pitch, but I've heard that is normal. The
roughing pump does change its sound periodically. Especially when
pumping down, it sounds like it is running rough. The scope didn't have
a maintenance contract at its former home, so I question how much
preventative maintenance was done. If I had to guess, I'd question if
the scope needs a rebuild. I've seen kits available. (Edwards E2M12 I
think.)

We can't exactly afford thousands of dollars to bring in a SEM tech.
Buying a rebuild kit for the roughing pump would be financially doable,
but I've heard this is a lot of work (Have any of you rebuilt a
multi-stage rotary vane pump? How difficult?), and I'd probably need an
offsite area to do the rebuild because our "microscopy lab" is in a
corner of a dusty workshop. (Yes, I know this isn't ideal for a SEM....)
If possible, I'd really like more of an indication that the roughing
pump is the problem before trying this.

Have any of you seen problems like this? Any thoughts on how to proceed?

Thanks,
Ryan

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Email: n00183308-at-ospreys.unf.edu
Name: Jason Saredy

Organization: University of North Florida

Title-Subject: [Filtered] JEOL JSM-35 Circuit Diagrams

Message: Hi all

I have circuit diagrams (for each component) and part list from a 35CF I was repairing years ago.
While the department ended up trashing the equipment, I still have diagrams. I have no idea whether
JEOL offers these in electronic formats, but I'd rather see the paper outside of a garbage heap.
Willing to eat mailing costs within the US, beautiful drawings in general and decent condition.

-Jason Saredy
N00183308-at-ospreys.unf.edu

PS for some reason the server keeps rejecting my email, school transferred services and domain names
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Email: jcsmtf-at-mail.missouri.edu
Name: Josh Schorp

Organization: University of Missouri

Title-Subject: [Filtered] Diffusion pump oil changing

Message: What is the proper way to change the oil in the diffusion pump on the Philips CM10/12? The
manual does not say how to release the vacuum in the pump. I removed the dipstick on the pump and
noticed that while the pressure was equalizing, most of the oil from the pump got sucked into the
tubes going to the buffer tank and the one to the rough pump. I know now that there was too much oil
in it, but is that the proper way to release the vacuum?

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Name: Philippe Pinard

Organization: RWTH Aachen University

Title-Subject: [Filtered] Monte Carlo Simulations for Electron Microscopy in Aachen (Germany)

Message: Dear fellow microscopists,

The Central Facility for Electron Microscopy of the RWTH Aachen University in collaboration with the
European Microbeam Analysis Society would like to invite you to the "Monte Carlo Simulations for
Electron Microscopy" workshop, that will be held at the RWTH Aachen University (Germany) from June
10th to 12th, 2014.

The aim of this workshop is to provide electron microscopists in-depth knowledge of Monte Carlo
simulation methods to assist them in their daily scientific work. Lectures from leading Monte Carlo
experts will cover both theoretical and practical aspects of current Monte Carlo programs. Tutorials
and live demonstrations will highlight common applications of Monte Carlo simulations to the field
of electron microscopy.

The registration for the workshop costs 150 euros for EMAS members or RWTH personnel and 200 euros
for other participants. The course is designed for any scientist (students, technicians,
researchers) having basic knowledge in EPMA, SEM and TEM.

For the full program, registration form and any other information about the workshop, please visit
our website:
http://www.gfe.rwth-aachen.de/montecarlo

Sincerely yours,
Philippe Pinard and Silvia Richter

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From affordable-meds14-at-telecom.kz Wed Apr 2 09:59:15 2014
Return-Path: {affordable-meds14-at-telecom.kz}
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Message-Id: {201404021459.s32ExB4l005618-at-ns.microscopy.com}

Hello Josh,
Bellow I add a copy of two paragraphs from the original 30 years old
manual for CM10/12. You have to follow all the steps.

----------------------------
6. 2. 1. ADMITTING AIR INTO THE PUMPING SYSTEM
Before air is let into the pumping system the OD pump should be allowed
to cool down for at least 1 hour after being switched off (either by
switching off the microscope or by removal of the OD pump fuse). Air
should first be admitted into the projection chamber (operating the
"cam air" softkey), then let air into the buffer tank and the diffusion
pump by very slowly and carefully loosening the dipstick 216 to 219,
Fig. 820.

Air is let into the preâ€”vacuum lines by opening valve V1
using the service key and the manual page. Air can also be admitted
into the pre- vacuum lines by removing Pirani P2. (Here should be P1, I
think.)
----------------------------

Best regards from Prague
Oldrich

--
Oldrich Benada
Institute of Microbiology AS CR, v.v.i.
Videnska 1083
142 20 Prague 4
Czech Republic

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} Title-Subject: [Filtered] Diffusion pump oil changing
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} Message: What is the proper way to change the oil in the diffusion
} pump on the Philips CM10/12? The manual does not say how to release
} the vacuum in the pump. I removed the dipstick on the pump and
} noticed that while the pressure was equalizing, most of the oil from
} the pump got sucked into the tubes going to the buffer tank and the
} one to the rough pump. I know now that there was too much oil in it,
} but is that the proper way to release the vacuum?
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8, 41 -- From benada-at-biomed.cas.cz Wed Apr 2 11:21:18 2014
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Email: kattymansouri-at-gmail.com
Name: Katayoun Mansouri

Organization: NCSU

Title-Subject: [Filtered] Precise measurement of particles from Rotary shadowing of freeze fracture
replicas

Message: I would like to measure the diameter of certain IMPs on protoplasmic fracture face (PF) of
a plant cell plasma membrane. I am fracturing at -160 degree C under a vacuum less than 10 to -7
mbar in a Cressington 308 freeze fracture machine. The platinum-carbon (Pt/C) is depositing from a
nominal 60 degree angle (the angle is relative to the specimen stage), and specimen is rotating at
maximum speed (the revolution per min is unknown but can be calculated). The rotary shadowing of
Pt/C is followed by carbon shadowing to stabilize the replica.
The deposited Pt/C is measured by a Quartz crystal and displayed on a Â“thickness monitorÂ”.
I would like to know you answers or comments about following questions:

1-With rotary shadowing what formula is used to calculate shadowed metal on a sphere particle?
2-How the angle that Pt/C is deposited from (e.g. 45 or 60 degree) incorporated to the formula?
3-Specimen rotation is used to uniform the shadowing on particle surface, however, dose it need to
be incorporated in the formula?
4-What is the formula for a unidirectional shadowing? Is it subtracting two times of the deposited
metal (read by Quartz crystal) from the diameter of shadowed particle?

Thanks,

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From mlequily-at-vnpt-hanoi.com.vn Wed Apr 2 21:47:59 2014
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Message-Id: {201404030247.s332lvhg026914-at-ns.microscopy.com}

Hi!

We have a Megaview II camera mounted on a Tecnai G20 instrument and are happy with it. Before SIS was bought by Olympus we got really good support. This is not a critic to Olympus, it is just that since then I never needed help anymore ;-)
Actually we never got any problem, I just needed help for optimization and the support was both friendly and efficient.
I find the original software better than the soft from FEI to control the camera.
I have no personnal interest but I can recommend it.

Stephane

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Email: see.wee.chee-at-gmail.com
Name: See Wee Chee

Title-Subject: [Filtered] Olympus TEM Cameras

Message: Hello everybody,

The side-mounted camera on our TEM has failed and we are looking for a replacement. We discovered
that Olympus is now in market with three side-mounted cameras. The specifications look pretty good
but we don't know of anyone that has one in service.

Has anyone here used an Olympus camera on their TEM before? Any feedback on performance, software
and support will be greatly appreciated. We are interested in the MegaView G2 and Veleta lines. Thanks!

See Wee

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Contents Retrieved from Microscopy Listserver Archives
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Greetings,

This is my first post to the list and while I have 40 years of microscopy experience I am new to the list so if my post is not in alignment with the rules I am very receptive to input so that I can be a productive list member.

Although it is not a TEM camera, we just put in a new Olympus research grade compound scope and elected to buy their higher end DP80 camera which was more costly than other options from third parties. That said, I have no regrets and the image quality is phenomenal. I am really glad that I elected to purchase it. This is a new system so I have no input on lifetime but I can say that I have never had such excellent images from a microscope before. Their service and support have been very good and the overall experience was very good. I would not hesitate to purchase another scope from Olympus and in fact we are getting ready to. If their TEM camera performs for you like the DP80 does for us I think you would be very happy. Hopefully, someone on the list with direct experience can provide feedback on the actual model you are looking at. You should also be able to work with one at a demo center which I would encourage you to do if you are not sure.

Sincerely,

James Neal-Kababick
Director
Flora Research Laboratories
An FDA and DEA Registered & Inspected Laboratory
Fellow AOAC International
Adjunct Faculty Bastyr University
Botanical Medicine Department
1-541-472-0980 phone
jimk-at-floraresearch.com
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From meds-affordable9-at-kes.ru Thu Apr 3 14:24:33 2014
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Greetings All,

I was hoping to get any information regarding immuno-gold labeling workshops/courses being offered this year in The U.S. I currently have quite a bit of experience in conventional biological TEM work but was hoping to expand my capacity to include immuno gold labeling. I have no experience with this technique and was hoping some sort of course or workshop might exist.

Kind Regards,

David Cugier, Associate Scientist II
Investigative Tox/Path-Preclinical Safety Division
Dept. R45M
AbbVie, Inc.
1 North Waukegan Road
North Chicago, Illinois 60064

Phone 847-938-6725
David.Cugier-at-Abbvie.com

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From pharmacy_discounted13-at-schaperpaint.com Fri Apr 4 21:17:49 2014
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Email: analytic-at-rawbw.com
Name: Margo Gill-Linscott

Organization: Northern California Society of Microscopy

Title-Subject: [Filtered] Spring Meeting

Message: Subject: [Filtered] Northern California Society for Microscopy Spring Meeting
April 17th

Northern California Society for Microscopy Spring Meeting

Thursday, April 17, 2014 6:00 PM to 9:00 PM
Please RSVP to info-at-ncsmicroscopy.org

Scotts of Palo Alto
#1 Town and Country Village
Palo Alto, California
http://scottsseafoodpa.com/

Featured Speakers Are:

Kent McDonald, PhD, Electron Microscopy Lab, UC Berkeley

Rapid Specimen Processing Methods for Cryofixed Samples

New methods for freeze substitution have been developed that
require only basic laboratory tools. Excellent freeze substitution results can be obtained in as
little as 90 min for cells of small volume. Rapid epoxy resin infiltration can be achieved by
centrifugation followed by polymerization at 100 C for 1.5Â–2 h. Total processing time from freezing
to blocks ready to section is about 6 h. Comparative results will be presented.

Ryan McGorty, PhD, UC San Francisco, School of Pharmacy

Three-dimensional super-resolution imaging of tissue samples

Single-molecule localization microscopy methods are often used to study samples at or very near the
coverslip surface with a spatial resolution of around 20nm. However, a number of challenges emerge
when trying to image even a few microns past the coverslip surface, particularly when trying to
accurately localize molecules in 3D. After first explaining the principle of the super-resolution
microscopy technique we use, I will discuss multiple methods we have recently developed that allow
us to image deeper into tissue samples.

No Cost to Current Members. $25 for Non-Members.

Bring your check for 2014 dues and enjoy a 3-course meal and great speakers.
Annual dues for regular members are *$40*; students, *$20*; and a corporate membership(which
includes one person) is *$100, *payable to *NCSM*

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Email: jcsmtf-at-mail.missouri.edu
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Organization: University of Missouri

Title-Subject: [Filtered] CM10 service key

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Title-Subject: [Filtered] Want to buy a Pico STM

Message: Does anyone have an Agilent or Molecular Imaging Pico STM they want to sell? If so, I have $$$

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Dear all,

A reminder that the registration deadline for our CEMOVIS course is
approaching (15.4.2014)

Best
Benoît Zuber

---------- Forwarded message ----------
X-from: {benoit.zuber.work-at-gmail.com}

Dear all,

We are pleased to announce

the 4th UniBe Practical Course on Cryo-Electron Microscopy of Vitreous
Sections (CEMOVIS, a.k.a cryoEM of frozen-hydrated sections)

11-13 August 2014 / 14-16 August 2014

Organiser : Benoît Zuber
Instructors: Benoît Zuber, Daniel Studer, Ioan Iacovache

The objective of the course is to teach the participants the practical
skills necessary to successfully apply CEMOVIS in their laboratories.
Essential background theory of CEMOVIS will be given and most of the
time will be spent practicing high-pressure freezing, cryo-sectioning,
and low-dose TEM imaging. 1 high-pressure freezing machine, 3
state-of-the-art cryo-ultramicrotomes and 1 cryo-electron microscope
will be dedicated to the course. Of special interest, participants
will learn to use the new tools that we developed to facilitate
cryosectioning (Studer et al 2014 J Struct Biol 185(1):125-8). The
3-day course is given twice, each time with a maximum of 4
participants so that they can be actively practicing at any time.

The course is intended for scientists whose research projects will
benefit from the use of CEMOVIS. Experience in either cryo-electron
microscopy or ultramicrotomy of resin-embedded specimens is a
prerequisite.

Information and registration:

http://www.ana.unibe.ch/events/cemovis/index.html

Registration deadline: 15.04.2014

Yours faithfully

Benoît Zuber

Prof. Benoît Zuber
Institute of Anatomy
University of Bern
Baltzerstrasse 2
CH-3000 Bern 9
Switzerland
Tel. +41 31 631 84 40
benoit.zuber-at-ana.unibe.ch
http://www.ana.unibe.ch/~exmo/

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REMINDER:

Dear Colleagues,
I appreciate if there are some dermatologist and skin researchers
using the Microscopy Listserver, who might find this information useful:

41st Annual Meeting of the SCUR (Society for Cutaneous Ultrastructure
Research ROME,ITALY; 2014) June 5-7,2014

NEW: 2nd Announcement

"VILLA MONDRAGONE", Monte Porzio Catone (RM, ITALY) JUNE 5-7, 2014
GENERAL TOPIC: "SKIN AGEING and CANCER: From ULTRASTRUCTURE to CLINIC"
INFORMATION (Flyer, all forms)
-at- http://www.scur.org/content/e1174/e1181/e1182/index_ger.html

EXTENDED DEADLINE for ABSTRACT SUBMISSION: Thursday, 10th April 2014

Call for papers (in dermatology, dermatopathology, skin research,
presenting results using all ancient and modern imaging techniques)

Registration and Accommodation information available.

The deadline to submit titles and abstracts is April 10th, 2014.
(late entries will be accepted on demand and need to be indicated by a
mail to the Local Organizers at
costanza-at-med.uniroma2.it; anapat-at-uniroma2.it; a.orlandi-at-uniroma2.it )

For more information, visit our website at:
http://www.scur.org/content/e1174/e1181/e1182/index_ger.html
or www.scur.org

Contact us (in case of requests for special information) by email
also to: W.Muss-at-salk.at

We look forward to seeing you at the 41st Ann. Meeting of the SCUR
in Monte Porzio Catone (RM), 20 km southeast of ROME city center
(FRASCATI, near airport CIAMPINO or FIUMICINO), a marvelous place
to stay!

NEW: Abstracts of 40th Annual Meeting of the SCUR = 6th Joint
Meeting of SCUR with SSSR, the Society for Skin Structure Research
(Japan)in SALZBURG, AUSTRIA, May 2013
-at- http://www.nature.com/jid/journal/v134/n4/index.html#Abstracts

Best regards,
Wolfgang Muss, PhD
........................................
SCUR Secretary
Wolfgang MUSS
EM-Lab, Univ. Inst. Pathology,
SALK-LK (Gen. Hospital) and PMU (Private Paracelsus Medical University) Salzburg
A-5020 SALZBURG-AUSTRIA

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From lseprupy-at-avangarddsl.ru Mon Apr 7 13:55:21 2014
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Greetings

This is a long shot, and may be of interest only to microscopists in the SF bay area.

Our EM technology training program is looking for individuals to serve on our advisory committee. We are required to have outside reviewers help us chart the direction and future of our program, we do this using an advisory committee.

We invite anyone interested in helping us do the right thing to join the committee. We have about two in person meetings on campus in Stockton each year, so the ability to be on campus for these meetings is a plus. However, even distant advisors can help us via email, skype, etc. We are particularly looking for a someone familiar with biological techniques, but everyone is welcome.

Please let me know if you are interested or need more information.

Thanks

Jon

Jonathan Krupp
Applied Science, Business & Technology
San Joaquin Delta College
5151 Pacific Ave.
Stockton, CA 95207
209-954-5284
jkrupp-at-deltacollege.edu

Find us on Facebook -at-
Electron Microscopy at SJ Delta College

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Email: veilcs-at-earthlink.net
Name: Ron Veil

Organization: Veil Electron Instrument Lab Customer Services

Title-Subject: [Filtered] RE: (Philips) CM10 service key

Message: the service key was given out to field service technicians when the CM was in production in
the '80s and '90s. The techs usually got these when they went to the factory training schools. The
service key had information burned onto an ePROM that one could use to manipulate the 'scope
"manually"; eg: to operate the vacuum sequence to check for leaks or proper operation. This is "old
technology" (~30 yrs) and both the ePROM series AND ePROM 'burners' are basically impossible to find.

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Title-Subject: [Filtered] SIS MegaViewII camera

Message: Does anyone have a old SIS MegaviewII camera that you would like to get rid of. I am
looking for parts to get the one we have working.

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Email: lucy.avati-at-sydney.edu.au
Name: Lucy Avati

Organization: University of Sydney

Title-Subject: [Filtered] Job vacancy: Light Microscopist / Senior Light Microscopist at the
University of Sydney

Message: Dear all,

The University of Sydney is currently seeking a Light Microscopist / Senior Light Microscopist. For
more information and to apply, visit sydney.edu.au/recruitment and search by the reference number
2440/1113A.

LIGHT MICROSCOPIST / SENIOR LIGHT MICROSCOPIST
SYDNEY MICROSCOPY & MICROANALYSIS
DEPUTY VICE-CHANCELLOR RESEARCH
REFERENCE NO. 2440/1113A
Â• Expertise in light/optical microscopy essential
Â• Full time fixed term to 31 December 2016, remuneration package: up to $95K p.a. which includes
leave loading and up to 17% super

Sydney Microscopy & Microanalysis (SMM) aims to be a world-leading facility for modern microscopy
and microanalysis, offering premier instruments, services and training to researchers from across
Sydney and around Australia, and providing strong leadership in the national and international
characterisation communities.

An exciting opportunity now exists to join SMM in the premier light optical microscopy position of
2014. In this role, you will work within SMMÂ’s existing facility and within the newly established
Cellular Imaging Facility at the UniversityÂ’s Charles Perkins Centre (CPC). The CPC is a new,
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From olommyey-at-nodosud.com.ar Tue Apr 8 13:54:13 2014
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Hi all,

I am searching for a method to preferentially stain lignin for SEM or TEM.
The object is to be able to determine if the lignin is present in
nano-size wood particles. Safranin can be used to stain lignin for LM but
does not contain a heavy element that will scatter electrons so that it
can be identified by EM.

Any ideas would be appreciated.

Debby

Debra Sherman
DS imaging LLC
Purdue Technology Center
1281 Win Hentschel Blvd
West Lafayette, IN 47906
E-mail: debby.sherman-at-dsimagingllc.com
www.dsimagingllc.com
Mobile: 765-418-8540

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Hi Debby,

If the lignin is at least partly exposed on the particle surface, you could use antibodies against lignin. With proper controls that will be a highly specific identification. See e.g. Visualizing Lignin Coalescence and Migration Through Maize Cell Walls Following Thermochemical Pretreatment, Bryon S. Donohoe, Stephen R. Decker, Melvin P. Tucker, Michael E. Himmel, Todd B. Vinzant
Biotechnology and Bioengineering, Vol. 101, No. 5, December 1, 2008

It does not have to be gold-labelled, but could be something like a solid phase immunoassay type of protocol: antibodies against lignin bound to a solid fase, which could be a film on a grid. Then incubate with the lignin suspension and see if you have specific binding on the film.

Will be happy to help. if required, with more details.

Cheers,

Jan

Jan Leunissen
Aurion
i: www.aurion.nl
Dept Geology
University of Otago
Dunedin, New Zealand

On 10/04/2014, at 1:37 pm, dsherman-at-purdue.edu wrote:

}
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} Hi all,
}
} I am searching for a method to preferentially stain lignin for SEM or TEM.
} The object is to be able to determine if the lignin is present in
} nano-size wood particles. Safranin can be used to stain lignin for LM but
} does not contain a heavy element that will scatter electrons so that it
} can be identified by EM.
}
} Any ideas would be appreciated.
}
} Debby
}
}
} Debra Sherman
} DS imaging LLC
} Purdue Technology Center
} 1281 Win Hentschel Blvd
} West Lafayette, IN 47906
} E-mail: debby.sherman-at-dsimagingllc.com
} www.dsimagingllc.com
} Mobile: 765-418-8540
}
}
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} 13, 31 -- Subject: lignin staining for TEM or SEM
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This was rejected when in html format, so reposting plain:

Hi Debby,

My colleague Colleen Macmillan, who has much experience staining for
lignin, suggests that the KMnO4 in the Maule stain may be electron-dense
enough. It certainly highlights cell membranes for TEM. I've pasted her
protocol below in case attachments won't go through:
_____________________________________________________
Mäule reaction lignin histology of plant sections

Samples: Easiest to place the (fresh) sections in a 48-well/multi-format
plate, then add and remove solutions from the sections using narrow-ended
plastic transfer pipettes.

Safety: Perform these reactions in a fume-hood, and wear nitrile gloves,
safety glasses, labcoat etc.

Mäule reaction protocol:
1. incubate sections (fresh is best) in KMnO-4 (1% aqueous) for 10 min
(if very thin sections then a couple of min is fine)
2. wash sections (ddH20)
3. acidify with conc. HCl (37%) for 1 min
4. wash again (ddH20)
5. incubate in NaHCO-3 (5% w/v; fresh) ~2-5 mins, or until colour
develops [then mount,
view, photograph etc.]

Indications:

red = syringyl lignin
brown = guaiacyl lignin

Based on pers. comm. from Armand Seguin + reference = Sibout, R., A.
Eudes, et al. (2005). "Cinnamyl alcohol dehydrogense C and D are the
primary genes involved in lignin biosynthesis in the floral stem of
Arabidopsis." Plant Cell 17(7): 2059-2076.

Mechanism: ŒChlorination of the syringyl nucleus leads to a pink
(lignifying cells) or red (lignified cells) color, whereas the guaiacyl
nucleus produces a light (lignifying cells) to dark (lignified cells)
brown color (Bland, 1966; Wardrop, 1981).¹ quote from US Patent Issued on
October 30, 2007, Inventor(s) Laigeng Li, Vincent Lee C. Chiang

Good luck,
cheers,
Rosemary White

Dr Rosemary White
CSIRO Plant Industry
GPO Box 1600
Canberra, ACT 2601
Australia

T 61 2 6246 5475
F 61 2 6246 5334
E rosemary.white-at-csiro.au

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Dear Debby,
dear all,

with regard to Rosemary White’s post I would like to point you to an indirect method to perhaps “localize” lignin in ultrathin sections (using UO2Ac-KMnO4-staining, see below)... There also has been made use of KMnO4 in the "double" staining.
Thanks to Rosemary for including the recipe for the Mäule-(Maeule)-Reaction!.

For your convenience I've copied and pasted part of the {Methods-section} , i.e. for TEM of Cell wall ultrastructure:
X-from:
American Journal of Botany 94(6): 912–925. 2007
REACTION TISSUE FORMATION AND STEM TENSILE MODULUS PROPERTIES IN WILD-TYPE AND P-COUMARATE-3-HYDROXYLASE
DOWNREGULATED LINES OF ALFALFA,MEDICAGO SATIVA (FABACEAE)

ANN M. PATTEN, MICHAE¨L JOURDES, ELVIE E. BROWN, MARIE-PIERRE LABORIE, LAURENCE B. DAVIN,
AND NORMAN G. LEWIS
Cf. also http://www.ncbi.nlm.nih.gov/pubmed/21636460 )

{ { Cell wall ultrastructure—
Identification of reaction vs. normal cell types was further confirmed using transmission electron microscopy (TEM).
Samples were harvested from a mature internode (IN20) from both WT and pC3H-I lines, as well as from branch tissue of black cottonwood as a control.
Tissues (ca. 5 mm2) were fixed in 2% paraformaldehyde and 1.25% glutaraldehyde in 50 mm piperazine-1,4-bis(2-ethanesulphonic acid) (PIPES) buffer (pH 7.2) overnight at 48C. Samples were dehydrated using a standard ethanol series, gradually infiltrated with LR White resin (London Resin, Reading, UK), and heat cured.
Thin sections were obtained using a diamond knife mounted to a Reichert Ultracut R ultramicrotome (Reichert-Jung GmbH, Heidelberg, Germany), and sections were mounted on formvar-coated 200-mesh nickel grids.

........Sections were stained with a 3:1 dilution of 4% (w/v) uranyl acetate and 1% (w/v) KMnO4 ........

and observed at 100 kV using a JEOL JEM-1200 EX transmission electron microscope (JEOL, Tokyo, Japan).} }

If you would like to get the .pdf for your personal use only(Copyright issue), please advise/request by reply to me.
Disclaimer: I haven't used the method so far, not working on wood or lignin localization....

Best wishes and regards,
Wolfgang

Von: Rosemary.White-at-csiro.au [mailto:Rosemary.White-at-csiro.au]
Gesendet: Donnerstag, 10. April 2014 07:26
An: Muß Wolfgang
Betreff: [Microscopy] Re: lignin for TEM/SEM? ==} KMnO4-in Mäule-reaction

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This was rejected when in html format, so reposting plain:

Hi Debby,

My colleague Colleen Macmillan, who has much experience staining for lignin, suggests that the KMnO4 in the Maule stain may be electron-dense enough. It certainly highlights cell membranes for TEM. I've pasted her protocol below in case attachments won't go through:
_____________________________________________________

Mäule reaction lignin histology of plant sections

Samples: Easiest to place the (fresh) sections in a 48-well/multi-format plate, then add and remove solutions from the sections using narrow-ended plastic transfer pipettes.

Safety: Perform these reactions in a fume-hood, and wear nitrile gloves, safety glasses, labcoat etc.

Mäule reaction protocol:
1. incubate sections (fresh is best) in KMnO4 (1% aqueous) for 10 min (if very thin sections then a couple of min is fine)
2. wash sections (ddH20)
3. acidify with conc. HCl (37%) for 1 min
4. wash again (ddH20)
5. incubate in NaHCO3 (5% w/v; fresh) ~2-5 mins, or until colour develops [then mount, view, photograph etc.]

Indications:

red = syringyl lignin
brown = guaiacyl lignin

Based on pers. comm. from Armand Seguin + reference = Sibout, R., A. Eudes, et al. (2005).
"Cinnamyl alcohol dehydrogense C and D are the primary genes involved in lignin biosynthesis in the floral stem of
Arabidopsis." Plant Cell 17(7): 2059-2076.

Mechanism: ŒChlorination of the syringyl nucleus leads to a pink (lignifying cells) or red (lignified cells) color, whereas the guaiacyl nucleus produces a light (lignifying cells) to dark (lignified cells) brown color (Bland, 1966; Wardrop, 1981).¹
quote from US Patent Issued on October 30, 2007, Inventor(s) Laigeng Li, Vincent Lee C. Chiang

Good luck,
cheers,
Rosemary White

Dr Rosemary White
CSIRO Plant Industry
GPO Box 1600
Canberra, ACT 2601
Australia

T 61 2 6246 5475
F 61 2 6246 5334
E rosemary.white-at-csiro.au
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Name: Richard Powell

Organization: Nanoprobes, Incorporated

Title-Subject: [Filtered] Re: [Microscopy] TEM ImmunoGold workshops or courses offered in the USA?

Message: Hello David:

One of the short courses at Microscopy & Microanalysis 2014, "Immunolabeling Technology for Light
and Electron Microscopy" will cover immunogold labeling in detail (disclosure: I will be one of the
instructors, and I work for Nanoprobes, a company that makes immunogold probes):

http://www.microscopy.org/MandM/2014/program/short_courses.cfm

Some of the other companies that make immunogold products also hold workshops and courses; I suggest
checking the web sites of manufacturers to see if there is an upcoming workshop or course near you.

Hope this helps,

Rick Powell
Nanoprobes, Incorporated

Greetings All,

I was hoping to get any information regarding immuno-gold labeling workshops/courses being offered
this year in The U.S. I currently have quite a bit of experience in conventional biological TEM
work but was hoping to expand my capacity to include immuno gold labeling. I have no experience
with this technique and was hoping some sort of course or workshop might exist.

Kind Regards,

David Cugier, Associate Scientist II
Investigative Tox/Path-Preclinical Safety Division
Dept. R45M
AbbVie, Inc.
1 North Waukegan Road
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Hi,

We have an opening for an Electron Microscopy service engineer in the Center for Cellular Imaging and NanoAnalytics (C-CINA) of the Biozentrum of the University of Basel (http://c-cina.org). We operate several TEM and SEM electron microscopes, including a FEI Titan Krios with a Gatan K2 Summit direct electron detector, FEI Polara, CM200FEG, T12-Autoloader, CM10, Helios FIB-SEM and a Quanta200FEG with a Gatan 3View.

We have an opening for an

=========================
EM Service Engineer
=========================

Your responsibilities will include the maintenance and repair of the electron microscopes. We do not have any factory maintenance contracts but may request additional help from the factory when needed. Responsibilities of this position are maintenance and repair of the electron microscopes including; problem diagnosis, ordering replacement parts from the factory, and repair. This includes: the Titan Krios Autoloader, cryo-stages, FEG and high voltage supplies, vacuum systems, mechanics, electronics and software backups.

In addition, upon interest, you would be welcome to participate in method development and research projects, if you were interested. We have an excellent mechanical and electronics workshop available, and are also developing methods in our group. Our research concerns high-resolution structure determination of biological molecules.

Salary and living conditions in Basel are attractive. The lab language is English.

Your profile
Experience in EM service for TEM, ideally also for SEM and FIB. Hardware repair experience.

We offer
This is a full-time position that is funded directly by the Rektorat of the University of Basel. It is a permanent position in a University setting in one of the most beautiful countries in Europe. We offer a welcoming working environment in an international and multidisciplinary group covering biochemistry, electron microscopy, structural biology, and data analysis. We are an equal opportunity employer.

Basel is an international city with people from 150 nations. Located at the border where three countries meet - Switzerland-Germany-France. It is Europe’s most important life sciences hub. Basel provides a high standard of living and a rich and varied cultural atmosphere.

For further information, please contact (CONFIDENTIALITY IS ASSURED):

Henning Stahlberg, Director
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Prof. for Structural Biology and Biophysics, Biozentrum,
WRO-1058, Mattenstrasse 26
University of Basel, CH-4058 Basel, Switzerland
Tel: +41-61-387 32 62 ; http://c-cina.org
mailto:Henning.Stahlberg-at-unibas.ch

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Dear Listers,

Just in case you missed last week's webinar on standardizing color
integrity in your brightfield color images and on your monitors using
the new ChromaCal system, the webinar is now available on line::
http://scientific.datacolor.com/watch-recording-apr-2-webinar-ensuring-color-integrity-datacolor-chromacal/?afftid=701G0000000tVqU

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Email: escanley-at-gmail.com
Name: Ellen Scanley

Organization: Southern Connecticut State University

Title-Subject: [Filtered] Flash Drive virus/malware protection

Message: Hi Listserve,

I am wondering what types of things people are
doing to make sure that their flash drives are
virus and malware free?

Our microscopes are not networked for cyber-security
purposes but we do use flash drives to download data
and sometimes to upload files to the microscope computers.

Thanks for suggestions !

Best Regards,
Ellen

Ellen Scanley, MD, PhD
Southern Connecticut State University
ConnSCU - Center for Nanotechnology
501 Crescent St.
New Haven, CT 06515

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17, 28 -- From microscopylistserver-noreply-at-microscopy.com Fri Apr 11 17:42:24 2014
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From express_drugstore9-at-tedpro.com Sat Apr 12 04:36:22 2014
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I would be hesitant in using flash drives. You could set up a system to scan them in another computer before plugging them into the microscope computer. However, even if you officially have a protocol in place to scan the drives before they go into the microscope computer, it wouldn't take but a single infection to ruin your day.

I think a better solution would be to attach the instrument computer to another computer that would serve as a firewall between the microscope and the broader network. Users could save their files to the intermediary and then use their method of choice to retrieve them. They could use a thumb drive or share them over the network. FEI, at least, uses such an approach for their microscopes and we have adopted the approach for other instrumentation running on old computers.

Warren

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Organization: Southern Connecticut State University

Title-Subject: [Filtered] Flash Drive virus/malware protection

Message: Hi Listserve,

I am wondering what types of things people are doing to make sure that their flash drives are virus and malware free?

Our microscopes are not networked for cyber-security purposes but we do use flash drives to download data and sometimes to upload files to the microscope computers.

Thanks for suggestions !

Best Regards,
Ellen

Ellen Scanley, MD, PhD
Southern Connecticut State University
ConnSCU - Center for Nanotechnology
501 Crescent St.
New Haven, CT 06515

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26, 35 -- From: "Straszheim, Warren E [BIOTC]" {wesaia-at-iastate.edu}
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26, 35 -- Subject: RE: [Microscopy] viaWWW:Flash Drive virus/malware protection
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From drugstore_affordable3-at-harrisnesbitt.com Sun Apr 13 04:37:28 2014
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In our facility, we have two dedicated, sterile flash drives to move data around within the facility, but only use a server or Google Drive for data leaving the facility. The best way to make sterile flash drives to to buy new drives, and then perform a long format on a computer with a fresh OS install.

In my experience, if users are bringing in their own drives to transfer data, it is inevitable that your equipment will become infected. To make our systems secure over the internet, the server is fire-walled to only allow access via the university subnet, and the routers are whitelisted to only allow access via ports 22, 80, and 443. We also have the internet browsers locked down to only allow access to Google Drive to keep people from surfing the internet on the computers.

We also have a wireless guest network on the router, such that users can still get the internet on their own devices, but cannot get access to the protected NAT.

We've had this system in place for over a year and have had very good success with it so far.

Hope this helps,
Ben Smith

Benjamin E. Smith, Ph.D.
Samuel Roberts Noble Microscopy Laboratory
University of Oklahoma
Norman, OK 73019
E-mail: benjamin.smith-at-ou.edu
Voice 405-325-4391
FAX 405-325-7619
http://www.microscopy.ou.edu/
________________________________________
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Name: Ellen Scanley

Organization: Southern Connecticut State University

Title-Subject: [Filtered] Flash Drive virus/malware protection

Message: Hi Listserve,

I am wondering what types of things people are
doing to make sure that their flash drives are
virus and malware free?

Our microscopes are not networked for cyber-security
purposes but we do use flash drives to download data
and sometimes to upload files to the microscope computers.

Thanks for suggestions !

Best Regards,
Ellen

Ellen Scanley, MD, PhD
Southern Connecticut State University
ConnSCU - Center for Nanotechnology
501 Crescent St.
New Haven, CT 06515

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Hello Ellen,
In our facility, we have microscope PC on a local network and one
other PC running Linux OS (Debian Wheezy). This PC is connected via
SAMBA to the microscope PC (read-only shared folders with guest's
images). Every user has a possibility to copy images from microscope PC
onto her/his flash drive on Linux box. No uploads to microscope PC are
permitted for normal user (guest).

Best regards Oldrich

--
Oldrich Benada
Institute of Microbiology AS CR, v.v.i.
Videnska 1083
142 20 Prague 4
Czech Republic

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} I am wondering what types of things people are
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} Our microscopes are not networked for cyber-security
} purposes but we do use flash drives to download data
} and sometimes to upload files to the microscope computers.
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} Thanks for suggestions !
}
} Best Regards,
} Ellen
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4, 43 -- From benada-at-biomed.cas.cz Mon Apr 14 09:23:55 2014
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OK, help me with this.

Does anyone have some sage advise about how microscopy and biotechnology can be integrated?

I have a microscopy training program, another nearby campus has a biotech program. We would like to find some common ground so our students can benefit from learning both.

This may seem like a silly question, after all, where would any science be without microscopes :).

But we are putting together a poster to show new students how these areas can overlap, so if you have any good ideas, send them my way.

Thanks

Jon

Jonathan Krupp
Applied Science, Business & Technology
San Joaquin Delta College
5151 Pacific Ave.
Stockton, CA 95207
209-954-5284
jkrupp-at-deltacollege.edu

Find us on Facebook -at-
Electron Microscopy at SJ Delta College

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Email: margaret.bisher-at-gmail.com
Name: Margaret Bisher

Organization: University of Pittsburgh School of Medicine

Title-Subject: [Filtered] Offer: Microscopy Books

Message: Trying to thin out my library and I was wondering if anyone would want some/all of these books?

Principle and Practice of Electron Microscope Operation
Low Temperature Methods in Biological Electron Microscopy
Electron Diffraction: An Introduction for Biologists
Design of the Electron Microscopy Laboratory
Image Analysis, Enhancement and Interpretation
- all of these are from the series: Practical Methods in Electron Microscopy

And I also have Vacuum Methods in Electron Microscopy, by Wilbur C. Bigelow.

The books are free, all I ask is that you pay the shipping costs.

Thank you, Margaret Bisher

University of Pittsburgh
School of Medicine
Department of Cell Biology
S346 Biomedical Science Tower
Pittsburgh, PA 15213

email: meb187-at-pitt.edu
phone: 412-648-9565

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Title-Subject: [Filtered] FlashDrive virus/malware protection

Message: Ellen,

We have installed USB Drive AntiVirus on our computers that scans flash drives and supposedly
prevents them from infecting the computers.

David

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Title-Subject: [Microscopy] Microscopy & Biotechnology question

Message: Jon,
Perhaps the following may serve as an inspiration to you and your colleagues:
http://blog.neuinfo.org/index.php/news-events/general-information/vasculature-morphogenesis-synopsis-of-three-related-articles-by-halina-witkiewicz-phil-oh-and-jan-schnitzer.

Regards,
Halina

} From: jkrupp-at-deltacollege.edu [jkrupp-at-deltacollege.edu]
Sent: Monday, April 14, 2014 2:11 PM
To: Witkiewicz, Halina

OK, help me with this.

Does anyone have some sage advise about how microscopy and biotechnology can be integrated?

I have a microscopy training program, another nearby campus has a biotech program. We would like to
find some common ground so our students can benefit from learning both.

This may seem like a silly question, after all, where would any science be without microscopes :).

But we are putting together a poster to show new students how these areas can overlap, so if you
have any good ideas, send them my way.

Thanks

Jon

Jonathan Krupp
Applied Science, Business & Technology
San Joaquin Delta College
5151 Pacific Ave.
Stockton, CA 95207
209-954-5284
jkrupp-at-deltacollege.edu

Find us on Facebook -at-
Electron Microscopy at SJ Delta College

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From drugs-express7-at-mail2moon.com Mon Apr 14 23:05:12 2014
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Jon,

Drosophila embryos.
One of our profs works on shrimp development - fertilization through
juveniles, and they use both molecular biology methods chasing the
relevant genes, Western blots and all that, and confocal microscopy to
see exactly where the genes are being expressed. Which cells and where
within the cells. Something molecular techniques don't tell you.
And, since he teaches the confocal class, he's used the same methods to
demonstrate the various homeobox genes in Drosophila embryos. Do the
molecular biology to get the information it provides, then image the
embroyos. Really nifty segmental patterns.
(Make "what does molecular biology/microscopy tell me?" and "how does
the one relate to the other?" part of the assignment. Don't tell them,
let them discover the relationships.)

Another does similar things with C. elegans. Two model organisms well
suited for class work, with lots of information Out There.

You could do similar things with cells in culture, following gene
expression through protein synthesis and transport between cellular
compartments. Run the techniques in parallel.

You're right, though - the techniques integrate well.

Phil

} OK, help me with this.
}
} Does anyone have some sage advise about how microscopy and biotechnology can be integrated?
}
} I have a microscopy training program, another nearby campus has a biotech program. We would like to find some common ground so our students can benefit from learning both.
}
} This may seem like a silly question, after all, where would any science be without microscopes :).
}
} But we are putting together a poster to show new students how these areas can overlap, so if you have any good ideas, send them my way.
}
} Thanks
}
} Jon
}
} Jonathan Krupp
} Applied Science, Business& Technology
} San Joaquin Delta College
} 5151 Pacific Ave.
} Stockton, CA 95207
} 209-954-5284
} jkrupp-at-deltacollege.edu
}
} Find us on Facebook -at-
} Electron Microscopy at SJ Delta College
--
Philip Oshel
Microscopy Facility Supervisor
Biology Department
024C Brooks Hall
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576

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Does anyone have a favorite freeze-substitution recipe that enhances the staining of the plasma membrane and tonoplast?
I usually use Osmium and UA in acetone. Is there something else I can add to the nasty cocktail to enhance the membranes?
thanks for any help,
Beth

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Email: zhouhw33-at-gmail.com
Name: Hongwen Zhou

Organization: Applied Materials

Title-Subject: [Filtered] Vacuum leak in JEOL ARM200F TEM

Message: We have a JEOL ARM200F TEM in our lab. The system has two vacuum-related issues, which lead
to only ~ 30% up time since we purchased it last year. The first one is that the chamber vacuum had
crashed at least five times when we were filling the liquid-N2 dewar. The second one is that the
samples are contaminated (by carbon) very quickly when we take images and/or do EELS/EDS analysis
under STEM mode. All samples are plasma-cleaned before inserting them into the ARM. And, we have not
observed such high contamination rate for the same samples in other TEMs. We detected two leaking
spots (by Helium leak test) on the goniometer and suspect those might be the root-cause of carbon
contamination. JEOL recommends us beam-shower the samples before STEM imaging/analysis, but that
destroys some of the samples and changes the structure of the materials in some cases.

I wonder if anyone had done any study on the correlation between carbon contamination (under STEM)
and low level vacuum leak before. Is it a pre-requisite to beam-shower the samples to obtain atomic
resolution images and high-quality EELS/EDS results? Thank you in advance for your input.

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Organization: Max Planck Institute of Biophysics

Title-Subject: [Filtered] E-mail to the list with job advertisement

Message: Dear Sir or Madam:

Could you please send the following job advertisement to the list?

Many thanks.

Yours sincerely,

Monika Hobrack
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Hi

I think you will have realised that you have an "O" ring problem on two
items that sit in the specimen area. It is extremely important to solve
these two problems before trying to characterise the vacuum system and the
contamination.

Another point, a leak in the specimen area may also cause etching of
specimens, reducing their cross section due to gas ionisation. For example
a holey carbon film may be reduced to the structure of a string vest in
minutes due to etching.

So, fix the problems that you have found, and the contamination problem may
well go away.

Good luck

Steve

Steve Chapman FRMS
Protrain for Consultancy and Courses in Electron Microscopy
Mobile +44 (0) 7711 606967 Fax +44 (0)1280 814007
www.emcourses.com

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Email: zhouhw33-at-gmail.com
Name: Hongwen Zhou

Organization: Applied Materials

Title-Subject: [Filtered] Vacuum leak in JEOL ARM200F TEM

Message: We have a JEOL ARM200F TEM in our lab. The system has two
vacuum-related issues, which lead to only ~ 30% up time since we purchased
it last year. The first one is that the chamber vacuum had crashed at least
five times when we were filling the liquid-N2 dewar. The second one is that
the samples are contaminated (by carbon) very quickly when we take images
and/or do EELS/EDS analysis under STEM mode. All samples are plasma-cleaned
before inserting them into the ARM. And, we have not observed such high
contamination rate for the same samples in other TEMs. We detected two
leaking spots (by Helium leak test) on the goniometer and suspect those
might be the root-cause of carbon contamination. JEOL recommends us
beam-shower the samples before STEM imaging/analysis, but that destroys some
of the samples and changes the structure of the materials in some cases.

I wonder if anyone had done any study on the correlation between carbon
contamination (under STEM) and low level vacuum leak before. Is it a
pre-requisite to beam-shower the samples to obtain atomic resolution images
and high-quality EELS/EDS results? Thank you in advance for your input.

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Dear All,
maybe there is someone out there before going into Easter holidays to share a bit of knowledge.
I just founds a strange detail on a specimen (the upper leaf surface of a cucumber plant):
See images here:
www.electronmicroscopy.info/customers/cucumber/index.htm
There is also a PDF with EDS data, but it is not very specific since I put a whole lot of Platinum on the surface because of a
nanoflight movie I did with this surface:
www.electronmicroscopy.info/customers/cucumber/cucumber_eds.pdf
A little bit of Sulfur and Ca is showing up, with Carbon.
Any idea what this might be?

Best wishes, happy Easter,
Stefan

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Hongwen;
We have had the column vacuum crashed on only one occasion, and that was when an inexperienced user filled the ACD to overfilling, and did not realize it was overflowing, so he poured and poured, which we believe froze the "O" ring that seals the top of the ACD. Aside from that, the column has never crashed. Contamination was a problem when we were heating the ACD every night. When we keep the ACD cold 24 hours a day, which can be easily done on the ARM, contamination from the column is eliminated. Contamination in a high current STEM (the ARM delivers ~10X the beam current density of a 2010F) is always a bigger problem than it is in conventional microscopes. We still see specimen-born contamination, and plasma cleaning does not seem to help as much as we would like. The best way to clean dirty specimens is baking them. We use the baking system that came with our NION UltraSTEM for samples that cannot be successfully cleaned by plasma cleaning.

John Mardinly, ASU

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

Email: zhouhw33-at-gmail.com
Name: Hongwen Zhou

Organization: Applied Materials

Title-Subject: [Filtered] Vacuum leak in JEOL ARM200F TEM

Message: We have a JEOL ARM200F TEM in our lab. The system has two vacuum-related issues, which lead to only ~ 30% up time since we purchased it last year. The first one is that the chamber vacuum had crashed at least five times when we were filling the liquid-N2 dewar. The second one is that the samples are contaminated (by carbon) very quickly when we take images and/or do EELS/EDS analysis under STEM mode. All samples are plasma-cleaned before inserting them into the ARM. And, we have not observed such high contamination rate for the same samples in other TEMs. We detected two leaking spots (by Helium leak test) on the goniometer and suspect those might be the root-cause of carbon contamination. JEOL recommends us beam-shower the samples before STEM imaging/analysis, but that destroys some of the samples and changes the structure of the materials in some cases.

I wonder if anyone had done any study on the correlation between carbon contamination (under STEM) and low level vacuum leak before. Is it a pre-requisite to beam-shower the samples to obtain atomic resolution images and high-quality EELS/EDS results? Thank you in advance for your input.

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realname - Naveen Rathi
Email - mpsnaveen0208-at-gmail.com
ORGANIZATION - Ecole Centrale de Nantes
EDUCATION - Graduate College
LOCATION - Nantes,France
SUBJECT_OF_QUESTION - 3D microscopy
QUESTION - Is there any robust and fastest way to segment the image in
3D using the Image j software by removing all the artifacts ?

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Email: thomas.burrage-at-dhs.gov
Name: Thomas G. Burrage

Organization: DHS S&T Plum Island Animal Disease Center

Title-Subject: [Filtered] EM Fellowship announcement

Message: Electron Microscopy Fellowship Appointment

Description:
The U.S. Department of Homeland Security (DHS) Science and Technology (S&T) Directorate is seeking
a motivated scientist (post-master’s or post-baccalaureate) with prior experience and skill sets
associated with electron microscopy techniques, or with a strong desire to pursue a scientific
career in electron microscopy projects associated with transboundary animal disease viruses (FMDV,
ASFV, CSFV). This fellowship is with the Transboundary Animal Disease Countermeasures Branch at the
Plum Island Animal Disease Center (PIADC) and is offered through the PIADC Research Participation
Program. The position is available immediately and is funded for 1 year, with the opportunity for
renewal.
PIADC is the only U.S. laboratory facility performing research, development and diagnosis of foreign
animal diseases of highest threat to the U.S. This critical national asset is located off the
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NY, and Old Saybrook, CT. One of the missions of PIADC is to develop technologies to help mitigate
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or deliberately introduced into the United States.
At PIADC, in a bio-safety level-3 (BSL-3) laboratory environment, the scientist will perform
laboratory procedures to visualize and analyze biopsy, tissue culture derived materials using
various microscopic procedures ranging from simple light microscopy to transmission electron
microscopy (TEM). Training will be provided. The scientist will prepare specimens and samples,
operate standard and specialized laboratory equipment, maintain tissue cultures, stay up-to-date
regarding technical developments, conduct literature searches on identified topics relevant to the
research, and follow and ensure adherence to strict safety procedures and safety checks.
Common methodologies employed include but are not limited to: tissue and cell preparation with
fixatives and embedding medias, ultrathin sectioning, paraffin-based cryomicrotomy and
immunohistochemistry; virus isolation, propagation and titration; preparation of vaccines; ELISAs;
evaluation of vaccine potency and efficacy in swine, cattle and mice; and use of several techniques
of recombinant DNA technology, real time PCR, protein expression in bacteria and in tissue culture,
light and fluorescence microscopy.
Qualifications:
The candidate must have demonstrated excellent oral and written communication skills and record
keeping skills, as the fellow will be working with agricultural select agents. The applicant must
be a U.S. Citizen and must be able to obtain a security clearance.
How to Apply:
Information about the application process and application forms are available on the program Web
site: http://www.orau.gov/piadc
For Additional Information:
For additional information about the PIADC Research Participation Program, please see
http://www.orau.gov/piadc.

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From popular-pharmacy7-at-childfocus.org Mon Apr 21 04:05:12 2014
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Email: fetterr-at-janelia.hhmi.org
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Organization: Howard Hughes Medical Institute

Title-Subject: [Filtered] Cressington 328C

Message: Has anyone used the Cressington 328EB Carbon unit to C coat support films on slot grids?
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Thanks for relevant answers.

Richard Fetter
Janelia Farm Research Campus
HHMI

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From pharmacy-discounted2-at-kmsf.org Mon Apr 21 10:14:09 2014
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Hello Fellow Microscopist,

A researcher in Florida is looking for a facility to perform freeze
fracture on E. coli membrane vesicles. The are 1) looking at the size but
2) more importantly, looking at the orientation if they membrane is right
side out or flipped inside out. They reference a 1974 paper in Journal
Bacteriology, Vol 117, No. 2.

My Core lab is more equipped to examine and model the membranes by
cryo-TEM and may try that but wondering if anyone is doing freeze fracture
anymore. And if so, where?

Best,
Karen Kelley
University of Florida
Interdisciplinary Center for Biotechnology Research

==============================Original Headers==============================
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Dr. John Rash from Colorado State University, is the best person I know who does freeze fracture. You can find more information from his website -

http://www.cvmbs.colostate.edu/rashlab/

Zhaojie Zhang
Director, Jenkins Microscopy Facility
University of Wyoming

-----Original Message-----
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Hello Fellow Microscopist,

A researcher in Florida is looking for a facility to perform freeze fracture on E. coli membrane vesicles. The are 1) looking at the size but
2) more importantly, looking at the orientation if they membrane is right side out or flipped inside out. They reference a 1974 paper in Journal Bacteriology, Vol 117, No. 2.

My Core lab is more equipped to examine and model the membranes by cryo-TEM and may try that but wondering if anyone is doing freeze fracture anymore. And if so, where?

Best,
Karen Kelley
University of Florida
Interdisciplinary Center for Biotechnology Research

==============================Original Headers==============================
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15, 34 -- From ZZhang-at-uwyo.edu Mon Apr 21 15:25:59 2014
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From pharmacy-discounted11-at-kam-telecom.ru Tue Apr 22 00:33:05 2014
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I have a client who wants to have a viscous liquid cryo-microtomed for TEM.

We don't have the equipment to provide such service and the nature of the liquid is still unknown to us. Does anyone who know who can provides this service so I can refer the inquiry to them?

Thanks........

Frank Karl
ARDL

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Anyone ever examine/analyze gunpowder or primer samples by
SEM? Any hints or cautions?

Thanks.

--
________
Larry D. Hanke, P.E.
Principal Engineer
Materials Evaluation and Engineering, Inc.
(763) 449-8870

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4, 16 -- From hanke-at-mee-inc.com Tue Apr 22 09:40:33 2014
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Not gunpowder, but all kinds of explosives powders - RDX, HMX, HNS, PETN and so forth. We make shaped charges for perforating oil and gas wells.

I usually gold coat to make the powders conductive and improve secondary electron yield, and use an aluminum stub for best thermal conductivity. I have a Lexan shield between me and the coater but have never, in the 20+ years I've been doing this, had any explosive decompose or deflagrate in the coater.

Could be wrong but I think most smokeless powders have a graphite coating, so they may already be conductive.

Hope this helps.

Regards,
Andrew T. Werner
Chief Metallurgist, Perforating
Schlumberger SRC
14910 Airline Road
Rosharon, TX 77583

"I Shoot the hippopotamus with bullets made of platinum,
because if I use leaden ones his hide is sure to flatten 'em"
The Bad Child's Book of Beasts - Hilaire Belloc

-----Original Message-----
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Sent: Tuesday, April 22, 2014 9:49 AM
To: Andrew Werner

Anyone ever examine/analyze gunpowder or primer samples by
SEM? Any hints or cautions?

Thanks.

--
________
Larry D. Hanke, P.E.
Principal Engineer
Materials Evaluation and Engineering, Inc.
(763) 449-8870

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19, 38 -- From werner1-at-slb.com Tue Apr 22 11:15:24 2014
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Hi Larry

I have not worked with what the customer called gun powder, but I guess the
material had similar properties.

I carried out a good deal of consultancy work with the then Nobel Explosives
Division of ICI (they have made explosives since the days of galleons!).
The specimens were mounted on a conventional stub using the smallest
possible quantity. There was no reason to coat the specimens. I used basic
specimen safety measures on a tungsten hairpin sourced instrument- low kV
( {2) lower emission current (30mA) and lower than normal spot sizes (i.e. a
spot size suitable for 40,000X when I was working below 10,000X). My safety
argument was that the amount of material was very small, and being contained
in a vacuum, I thought I was pretty safe.

The client was frightened to carry out the work themselves but were happy
with the results I provided for then on several 3 day visits.

Good luck

Steve

Steve Chapman FRMS
Protrain for Consultancy and Courses in Electron Microscopy
Mobile +44 (0) 7711 606967 Fax +44 (0)1280 814007
www.emcourses.com

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Anyone ever examine/analyze gunpowder or primer samples by SEM? Any hints or
cautions?

Thanks.

--
________
Larry D. Hanke, P.E.
Principal Engineer
Materials Evaluation and Engineering, Inc.
(763) 449-8870

==============================Original Headers==============================
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Title-Subject: [Filtered] CM10 error noise

Message: Today I just installed our newly rebuilt diffusion pump on our cm10. I ran the vacuum for
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Contents Retrieved from Microscopy Listserver Archives
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Dear Hongbing,

Your supervisor is quite right to warn you of the perils of using
Fourier transforms. I've seen some terrible abuse of FFTs, even in the
highest impact factor journals where the refereeing should have been
better.

I believe your request relates to measuring displacements and rotations
of the crystal lattices either side of an interface. Probably the best
article I would refer you to is Hytch, Snoeck & Kilaas (Ultramicroscopy
74 (1998) 131â€”146) who coined the term geometric phase analysis (GPA -
not to be confused with geometric phases in quantum mechanics). They
explain how to use the the FFT of a HRTEM image to recover information
about the displacement, R(x,y), and rotation, omega(x,y), of the lattice
from each of the peaks in the FFT. The strains and shears associated
with interfaces and steps can be seen as phase ramps. Differentiating
these will give local strains and shears.

There are certain caveats to using this technique however, which Martin
Hytch & Tobias Plamann address in a later paper (Ultramicroscopy 87
(2001) 199â€“212). The problem is that, in some crystals, the peaks (and
troughs) in a lattice image do not necessarily track the positions of
the atomic columns. This occurs if there are a) strong thickness
gradients, b) non-centrosymmetric crystals. For both, the local beam
tilt (if using a large condenser aperture) and crystal tilts (buckling
due to thin-film stresses) lead to the lattice fringes shifting across
the unit cell leading to phantom strains (in the case of centrosymmetric
crystals the lattice fringes shift by a factor of pi radians and the
peaks become troughs and vice versa).

From a practical point of view you need good quality HRTEM images so I
would recommend using small condenser apertures and, if possible, energy
filtering the image with a Gatan Imaging Filter or in-column filter.

Analysis using GPA can be done either using post-analysis software, e.g.
MatLab (I tend to use IDL), or by using a Digital Micrograph script that
can be purchased from HREM Research (www.hremresearch.com).*

I hope that helps.

Yours, Jon

Note: I have no links, commercial or otherwise, with Gatan, MatLab, IDL
or HREM Research. These are recommendations based on my past
experiences.

P.S. If you need a pdf of the articles I refer to, I can send you an
electronic copy. Please do ask.

} Title-Subject: [Filtered] High resolution TEM inverse fast Fourier
} transformation
}
} Message: Dear all,
} I am a PhD student in material science from Queen's University in
} Canada. I am trying to use HRTEM
} to characterize the grain boundaries and phase interfaces and
} dislocations generated after
} deformation in Zr alloy. I have captured some HRTEM images but it is
} very hard to interpret them. My
} supervisor told me that it should be very careful to explain them. And
} as it says in textbook
} written by William, we should use simulation software to interpret the
} image. However, in open
} literature, inverse fast Fourier transformation(IFFT) was widely used
} to index the dislocations and
} misfits between two phases. And based on my own experience, the IFFT
} image is related to the size
} and position of the mask which was applied. Could any one give me any
} idea about the advantages and
} disadvantages of IFFT method to analyze the HRTEM data. My
} appreciation for your suggestion and
} discussion.
} Best Regards
} Hongbing Yu

==============================Original Headers==============================
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From meds_popular1-at-bashtel.ru Wed Apr 23 10:10:15 2014
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I would be happy to be corrected if I was wrong but I always thought that an explosion was a very fast combustion.
And that a combustion needs oxygen, which is by definition absent from a high vacuum environment.
Can I convince somebody with my argument?

Stephane

X-from: "hanke-at-mee-inc.com" {hanke-at-mee-inc.com}
To: nizets2-at-yahoo.com
Sent: Tuesday, April 22, 2014 4:44 PM

Anyone ever examine/analyze gunpowder or primer samples by
SEM? Any hints or cautions?

Thanks.

--
________
Larry D. Hanke, P.E.
Principal Engineer
Materials Evaluation and Engineering, Inc.
(763) 449-8870

==============================Original Headers==============================
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13, 46 -- From: Stephane Nizet {nizets2-at-yahoo.com}
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From best_drugs3-at-tpnet.pl Thu Apr 24 06:35:53 2014
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Hi Stephane

Yes I 100% agree and you will see that I took this into account in my email.
I see no reason to go into fancy techniques when the simplest of procedures
will do the job. I did outline those that I had used without a problem
during my 6 days of investigations.

I did not carry out EDX and understand that if I had needed to do so I may
have required higher than the 2kV that I had used.

Kind regards

----------------------------------------------------------------------------
---------------------------------------
I would be happy to be corrected if I was wrong but I always thought that an
explosion was a very fast combustion.
And that a combustion needs oxygen, which is by definition absent from a
high vacuum environment.
Can I convince somebody with my argument?

Stephane

X-from: "hanke-at-mee-inc.com" {hanke-at-mee-inc.com}
To: nizets2-at-yahoo.com
Sent: Tuesday, April 22, 2014 4:44 PM

Anyone ever examine/analyze gunpowder or primer samples by
SEM? Any hints or cautions?

Thanks.

--
________
Larry D. Hanke, P.E.
Principal Engineer
Materials Evaluation and Engineering, Inc.
(763) 449-8870

==============================Original Headers==============================
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Contents Retrieved from Microscopy Listserver Archives
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Hi Stephane,

I believe that most explosives contain their own oxidizers to enable the
rapid reaction. Hence the presence of peroxide in many of the homemade
concoctions. Anyone want to clarify the issue?

Cheers,
Henk

On 4/24/2014 7:35 AM, nizets2-at-yahoo.com wrote:
}
}
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}
} I would be happy to be corrected if I was wrong but I always thought that an explosion was a very fast combustion.
} And that a combustion needs oxygen, which is by definition absent from a high vacuum environment.
} Can I convince somebody with my argument?
}
} Stephane
}
}
}
} X-from: "hanke-at-mee-inc.com" {hanke-at-mee-inc.com}
} To: nizets2-at-yahoo.com
} Sent: Tuesday, April 22, 2014 4:44 PM
} Subject: [Microscopy] SEM/EDS of Gunpowder
}
}
}
}
}
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
} To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
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} ----------------------------------------------------------------------------
}
} Anyone ever examine/analyze gunpowder or primer samples by
} SEM? Any hints or cautions?
}
} Thanks.
}

--

Hendrik O. Colijn

*C*enter for *E*lectron *M*icroscopy & *A*nalysi*S*

1305 Kinnear Rd, Suite 100, Columbus, OH 43212

colijn.1-at-osu.edu {mailto:colijn.1-at-osu.edu} 614.643.3458 Office

cemas.osu.edu

"Time is that quality of nature which keeps things from happening all at
once. Lately it doesn't seem to be working."

==============================Original Headers==============================
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That is my understanding as well. In an explosion, there is not enough time to rely upon mass transfer of oxygen to fuel the reaction. The reaction is a rearrangement of atoms already present.

The question will be what energy threshold needs to be exceeded to start the reaction. Low voltages and beam currents are a good approach. High explosives may be safer because they need quite a kick from a detonator to get started.

Of course, it is also helpful to limit the material examined so if something goes off it won't damage the instrument.

I'd be interested if someone could address those issues.

-----Original Message-----
X-from: colijn.1-at-osu.edu [mailto:colijn.1-at-osu.edu]
Sent: Thursday, April 24, 2014 7:16 AM

Hi Stephane,

I believe that most explosives contain their own oxidizers to enable the rapid reaction. Hence the presence of peroxide in many of the homemade concoctions. Anyone want to clarify the issue?

Cheers,
Henk

On 4/24/2014 7:35 AM, nizets2-at-yahoo.com wrote:
}
}
} I would be happy to be corrected if I was wrong but I always thought that an explosion was a very fast combustion.
} And that a combustion needs oxygen, which is by definition absent from a high vacuum environment.
} Can I convince somebody with my argument?
}
} Stephane
}
} X-from: "hanke-at-mee-inc.com" {hanke-at-mee-inc.com}
} To: nizets2-at-yahoo.com
} Sent: Tuesday, April 22, 2014 4:44 PM
} Subject: [Microscopy] SEM/EDS of Gunpowder
}
}
}
}
}
} ----------------------------------------------------------------------
} ------ 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
} ----------------------------------------------------------------------
} ------
}
} Anyone ever examine/analyze gunpowder or primer samples by SEM? Any
} hints or cautions?
}
} Thanks.
}

--

Hendrik O. Colijn

*C*enter for *E*lectron *M*icroscopy & *A*nalysi*S*

1305 Kinnear Rd, Suite 100, Columbus, OH 43212

colijn.1-at-osu.edu {mailto:colijn.1-at-osu.edu} 614.643.3458 Office

cemas.osu.edu

"Time is that quality of nature which keeps things from happening all at once. Lately it doesn't seem to be working."

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Stephane,

Combustion does require fuel and oxidizer. However, explosives and propellants such as smokeless powder are molecules containing hydrogen, carbon, nitrogen, and oxygen in a relatively stable configuration, that can be disturbed by energy input (shock, heat etc.) an decompose into carbon dioxide / monoxide, water vapor, and nitrogen.

That is, these compounds contain both fuel and oxidizer, and the explosion is essentially the reconfiguration of the components into more stable compounds, with a concomitant release of energy (heat and noise).

If you google TNT you will find that it is a 6 carbon benzene ring with one hydrogen, one CH3 group, and three NO2 groups. The reaction can be written (please excuse lack of subscripts):

C7H5N2O6 ---} 1.5 N2 + 2.5 H2O + 3.5 CO + 3.5 C. If it had a little more oxygen the last CO and C would be fully oxidized to CO2.

Anyway, it takes a "spark" - some unstabilizing input - to kick off the reaction. And since this has little to do with microscopy I'll shut up now.

Regards,
Andrew

-----Original Message-----
X-from: nizets2-at-yahoo.com [mailto:nizets2-at-yahoo.com]
Sent: Thursday, April 24, 2014 6:43 AM
To: Andrew Werner

I would be happy to be corrected if I was wrong but I always thought that an explosion was a very fast combustion.
And that a combustion needs oxygen, which is by definition absent from a high vacuum environment.
Can I convince somebody with my argument?

Stephane

X-from: "hanke-at-mee-inc.com" {hanke-at-mee-inc.com}
To: nizets2-at-yahoo.com
Sent: Tuesday, April 22, 2014 4:44 PM

Anyone ever examine/analyze gunpowder or primer samples by
SEM? Any hints or cautions?

Thanks.

--
________
Larry D. Hanke, P.E.
Principal Engineer
Materials Evaluation and Engineering, Inc.
(763) 449-8870

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Contents Retrieved from Microscopy Listserver Archives
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I've never been afraid of explosive damage, but the idea of releasing those potentially corrosive gases and moisture in my SEM has always given me the heebie-jeebies (surprisingly, the spell checker has the correct version of heebie-jeebies... but it didn't have microtome go figure.)

Stay safe.........
Frank

-----Original Message-----
X-from: wesaia-at-iastate.edu [mailto:wesaia-at-iastate.edu]
Sent: Thursday, April 24, 2014 9:54 AM
To: Frank Karl

That is my understanding as well. In an explosion, there is not enough time to rely upon mass transfer of oxygen to fuel the reaction. The reaction is a rearrangement of atoms already present.

The question will be what energy threshold needs to be exceeded to start the reaction. Low voltages and beam currents are a good approach. High explosives may be safer because they need quite a kick from a detonator to get started.

Of course, it is also helpful to limit the material examined so if something goes off it won't damage the instrument.

I'd be interested if someone could address those issues.

-----Original Message-----
X-from: colijn.1-at-osu.edu [mailto:colijn.1-at-osu.edu]
Sent: Thursday, April 24, 2014 7:16 AM

Hi Stephane,

I believe that most explosives contain their own oxidizers to enable the rapid reaction. Hence the presence of peroxide in many of the homemade concoctions. Anyone want to clarify the issue?

Cheers,
Henk

On 4/24/2014 7:35 AM, nizets2-at-yahoo.com wrote:
}
}
} I would be happy to be corrected if I was wrong but I always thought that an explosion was a very fast combustion.
} And that a combustion needs oxygen, which is by definition absent from a high vacuum environment.
} Can I convince somebody with my argument?
}
} Stephane
}
} X-from: "hanke-at-mee-inc.com" {hanke-at-mee-inc.com}
} To: nizets2-at-yahoo.com
} Sent: Tuesday, April 22, 2014 4:44 PM
} Subject: [Microscopy] SEM/EDS of Gunpowder
}
}
}
}
}
} ----------------------------------------------------------------------
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} of America To Subscribe/Unsubscribe --
} http://www.microscopy.com/MicroscopyListserver
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}
} Anyone ever examine/analyze gunpowder or primer samples by SEM? Any
} hints or cautions?
}
} Thanks.
}

--

Hendrik O. Colijn

*C*enter for *E*lectron *M*icroscopy & *A*nalysi*S*

1305 Kinnear Rd, Suite 100, Columbus, OH 43212

colijn.1-at-osu.edu {mailto:colijn.1-at-osu.edu} 614.643.3458 Office

cemas.osu.edu

"Time is that quality of nature which keeps things from happening all at once. Lately it doesn't seem to be working."

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

Gunpowder - Its Composition

Gunpowder ordinarily consists of three components-potassium nitrate, also called saltpeter (KNO3), charcoal (C), and sulfur (S). Proportions of these components may vary, and typically are 65 to 75 percent potassium nitrate, 15 to 20 percent charcoal, and 10 to 15 percent sulfur.

Interestingly, stoichiometrically,1 the reaction proportions are 84 to 8 to 8. The reaction is considered to generally be,

10 KNO3 + 8 C + 3 S â†’ 2 K2CO3 + 3 K2SO4 + 6 CO2â†‘ + 5 N2â†‘

Gunpowder - What Produces the Bang?

Most chemical reactions do not include a bang. Why does the ignition of gunpowder-after all, simply a chemical reaction-do so? It is because of the sudden generation of pressure within an enclosed space. Gunpowder in a closed tube creates enough pressure it violently shatters the container (perhaps made of steel) and generates a "shock wave."2 How?

The chemical reactants-all solid-do not take up much volume. On the other hand, the products generated include 11 parts of carbon dioxide and nitrogen-gases. Now the solid products are slightly less in volume than the solid reactants, so it might seem the pressure should be decreasing, not increasing. However, the volume of the gas products immensely greater, since the atoms in a gas are very far apart. It is the sudden conversion of a small amount of solid material into gas that produces tremendous pressure--suddenly bursting the container-that produces the characteristic explosion with its associated Bang! Anyone who has observed a fireworks display knows that at the end of a beautiful aerial flower his ears may be assaulted by an occasional series of sharp Bangs. Bang-bang-bang-bang-bang-BANG!

Regards,
Steve

Steve Buckingham
Surface Scientist
Surface Science
____________________________________________________________________

Novelis Global Research & Technology Center
1950 Vaughn Road
Kennesaw, GA 30144

Cell:Â Â Â 770-547-3381
Office: 770-795-6767
steve.buckingham-at-novelis.com
www.novelis.com

Not just aluminum, Novelis Aluminum.

-----Original Message-----
X-from: werner1-at-slb.com [mailto:werner1-at-slb.com]
Sent: Thursday, April 24, 2014 10:09 AM
To: Stephen Buckingham

Stephane,

Combustion does require fuel and oxidizer. However, explosives and propellants such as smokeless powder are molecules containing hydrogen, carbon, nitrogen, and oxygen in a relatively stable configuration, that can be disturbed by energy input (shock, heat etc.) an decompose into carbon dioxide / monoxide, water vapor, and nitrogen.

That is, these compounds contain both fuel and oxidizer, and the explosion is essentially the reconfiguration of the components into more stable compounds, with a concomitant release of energy (heat and noise).

If you google TNT you will find that it is a 6 carbon benzene ring with one hydrogen, one CH3 group, and three NO2 groups. The reaction can be written (please excuse lack of subscripts):

C7H5N2O6 ---} 1.5 N2 + 2.5 H2O + 3.5 CO + 3.5 C. If it had a little more oxygen the last CO and C would be fully oxidized to CO2.

Anyway, it takes a "spark" - some unstabilizing input - to kick off the reaction. And since this has little to do with microscopy I'll shut up now.

Regards,
Andrew

-----Original Message-----
X-from: nizets2-at-yahoo.com [mailto:nizets2-at-yahoo.com]
Sent: Thursday, April 24, 2014 6:43 AM
To: Andrew Werner

I would be happy to be corrected if I was wrong but I always thought that an explosion was a very fast combustion.
And that a combustion needs oxygen, which is by definition absent from a high vacuum environment.
Can I convince somebody with my argument?
Â
Stephane
Â
Â

X-from: "hanke-at-mee-inc.com" {hanke-at-mee-inc.com}
To: nizets2-at-yahoo.com
Sent: Tuesday, April 22, 2014 4:44 PM

Anyone ever examine/analyze gunpowder or primer samples by
SEM? Any hints or cautions?

Thanks.

--
________
Larry D. Hanke, P.E.
Principal Engineer
Materials Evaluation and Engineering, Inc.
(763) 449-8870

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Well, I said I'd shut up but a couple of additional points:

You are right, no time for mass transfer, the explosive molecule is broken down and rearranged into smaller, simpler molecules with the release of heat.

The energy threshold is sticky - it is why we have rules about cutting detonating cord with a very sharp blade and not "sawing" on it - friction can set it off, but it has to be enough, and concentrated/localized.

High explosives can go "low order" - they can deflagrate instead of detonate. High order detonation means the reaction front moves faster than the sound speed in the material; low order means the shock wave (sound) can outrun the reaction. High order puts the energy out faster/sooner. But high explosives aren't necessarily harder to initiate than low.

You are absolutely right about limiting the amount of energetic material, for a couple of reasons. One it total energy involved - deflagration of a few milligrams will contaminate the vacuum system but not damage the microscope chamber. The other is what is called "run up to detonation" and (roughly), works like - starting one spot burning, the reaction is relatively slow, but picks up speed as heat accumulates and finally gets fast enough to outrun the sound wave - it goes high order. Limiting the amount of material obviates this possibility.

Look, I'm a Metallurgist who works with explosives people and happens to use microscopes - not really a Microscopist or Shock Physicist or Chemical Engineer. But explosives are fascinating and useful - we use a lot more in mining and oilfield than they do in military applications every year. So, if anyone is interested in them, get Paul Cooper's book (with Stan Kurowski) Introduction to the Technology of Explosives - ISBN 0-471-18635-X - it is very accessible and fascinating.

Regards,
Andrew

-----Original Message-----
X-from: wesaia-at-iastate.edu [mailto:wesaia-at-iastate.edu]
Sent: Thursday, April 24, 2014 8:53 AM
To: Andrew Werner

That is my understanding as well. In an explosion, there is not enough time to rely upon mass transfer of oxygen to fuel the reaction. The reaction is a rearrangement of atoms already present.

The question will be what energy threshold needs to be exceeded to start the reaction. Low voltages and beam currents are a good approach. High explosives may be safer because they need quite a kick from a detonator to get started.

Of course, it is also helpful to limit the material examined so if something goes off it won't damage the instrument.

I'd be interested if someone could address those issues.

-----Original Message-----
X-from: colijn.1-at-osu.edu [mailto:colijn.1-at-osu.edu]
Sent: Thursday, April 24, 2014 7:16 AM

Hi Stephane,

I believe that most explosives contain their own oxidizers to enable the rapid reaction. Hence the presence of peroxide in many of the homemade concoctions. Anyone want to clarify the issue?

Cheers,
Henk

On 4/24/2014 7:35 AM, nizets2-at-yahoo.com wrote:
}
}
} I would be happy to be corrected if I was wrong but I always thought that an explosion was a very fast combustion.
} And that a combustion needs oxygen, which is by definition absent from a high vacuum environment.
} Can I convince somebody with my argument?
}
} Stephane
}
} X-from: "hanke-at-mee-inc.com" {hanke-at-mee-inc.com}
} To: nizets2-at-yahoo.com
} Sent: Tuesday, April 22, 2014 4:44 PM
} Subject: [Microscopy] SEM/EDS of Gunpowder
}
}
}
}
}
} ----------------------------------------------------------------------
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}
} Anyone ever examine/analyze gunpowder or primer samples by SEM? Any
} hints or cautions?
}
} Thanks.
}

--

Hendrik O. Colijn

*C*enter for *E*lectron *M*icroscopy & *A*nalysi*S*

1305 Kinnear Rd, Suite 100, Columbus, OH 43212

colijn.1-at-osu.edu {mailto:colijn.1-at-osu.edu} 614.643.3458 Office

cemas.osu.edu

"Time is that quality of nature which keeps things from happening all at once. Lately it doesn't seem to be working."

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colijn.1-at-osu.edu wrote:
}
}
} I believe that most explosives contain their own oxidizers to enable the
} rapid reaction. Hence the presence of peroxide in many of the homemade
} concoctions. Anyone want to clarify the issue?
}
}

A gunpowder explosion is a deflagration (rapid burning, yes, but
with the oxidizer in the composition) instead of a detonation,
which is what defines "high explosives". If you google those
two terms, you'll get a complete explanation.

Rick Mott

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Steve,

Exactly! This is what is known these days as "black powder" and was originally known as gunpowder before the advent of smokeless powder. Black powder is an intimate mixture of the three components, mixed, dampened, granulated, and dried.

Modern small arms propellants (smokeless powder) are generally nitrocellulose based (there are single base and double base powders, the double base contain some nitroglycerine), granulated, and coated with deterrent coatings that encourage progressive burning - controlled deflagration - to tailor the pressure-time curve to the projectile/barrel length combination at hand.

You can make nitrocellulose easily by nitrating cotton or paper with nitric and sulfuric acids. Note that this is something to do in controlled conditions, with proper cooling and safeguards - but my 7th grade Science Teacher, Miss Wyle, showed us and actually may be the person ultimately responsible for the course my life has taken (she was great).

Anyway, I wish it were Friday and I could continue this, but I've got to get back to work.

Regards,
Andrew

-----Original Message-----
X-from: Stephen Buckingham [mailto:steve.buckingham-at-novelis.com]
Sent: Thursday, April 24, 2014 9:11 AM
To: Andrew Werner
Cc: microscopy-at-microscopy.com

Stephane,

Combustion does require fuel and oxidizer. However, explosives and propellants such as smokeless powder are molecules containing hydrogen, carbon, nitrogen, and oxygen in a relatively stable configuration, that can be disturbed by energy input (shock, heat etc.) an decompose into carbon dioxide / monoxide, water vapor, and nitrogen.

That is, these compounds contain both fuel and oxidizer, and the explosion is essentially the reconfiguration of the components into more stable compounds, with a concomitant release of energy (heat and noise).

If you google TNT you will find that it is a 6 carbon benzene ring with one hydrogen, one CH3 group, and three NO2 groups. The reaction can be written (please excuse lack of subscripts):

C7H5N2O6 ---} 1.5 N2 + 2.5 H2O + 3.5 CO + 3.5 C. If it had a little more oxygen the last CO and C would be fully oxidized to CO2.

Anyway, it takes a "spark" - some unstabilizing input - to kick off the reaction. And since this has little to do with microscopy I'll shut up now.

Regards,
Andrew

-----Original Message-----
X-from: nizets2-at-yahoo.com [mailto:nizets2-at-yahoo.com]
Sent: Thursday, April 24, 2014 6:43 AM
To: Andrew Werner

I would be happy to be corrected if I was wrong but I always thought that an explosion was a very fast combustion.
And that a combustion needs oxygen, which is by definition absent from a high vacuum environment.
Can I convince somebody with my argument?
Â
Stephane
Â
Â

X-from: "hanke-at-mee-inc.com" {hanke-at-mee-inc.com}
To: nizets2-at-yahoo.com
Sent: Tuesday, April 22, 2014 4:44 PM

Anyone ever examine/analyze gunpowder or primer samples by
SEM? Any hints or cautions?

Thanks.

--
________
Larry D. Hanke, P.E.
Principal Engineer
Materials Evaluation and Engineering, Inc.
(763) 449-8870

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werner1-at-slb.com wrote:
}
}
} Modern small arms propellants (smokeless powder) are generally nitrocellulose based (there are single base and double base powders, the double base contain some nitroglycerine), granulated, and coated with deterrent coatings that encourage progressive burning - controlled deflagration - to tailor the pressure-time curve to the projectile/barrel length combination at hand.
}
} You can make nitrocellulose easily by nitrating cotton or paper with nitric and sulfuric acids. Note that this is something to do in controlled conditions, with proper cooling and safeguards - but my 7th grade Science Teacher, Miss Wyle, showed us and actually may be the person ultimately responsible for the course my life has taken (she was great).
}
}

Right, sorry, I was thinking about black powder, not modern
smokeless powder which is what "gunpowder" currently means.
Andrew was typing his more detailed explanations while I was
still sending mine.

This is a bit OT, but:

http://explosives.mst.edu/media/academic/explosives/documents/camp/2014ExplosivesCamp.pdf

if you have a kid 16 or older (or if you're young enough to pass
for a high-school kid, which definitely excludes me), check this out!

Rick Mott (guilty of making NI3 and other noisy fun stuff as
a teenager)

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Hi All

In relation to the comments about column contamination, the fumes from the
"explosive" instruments pump were disgusting. A column clean, new filters,
and fresh fluids in the pumps, as expected, made no difference. This column
was in real trouble after years of working with explosives.

The best solution we could use to reduce "operator" contamination was to
dump the RP outlet to the outside world; sorry greens!

Regards

Steve

Steve Chapman FRMS
Protrain for Consultancy and Courses in Electron Microscopy
Mobile +44 (0) 7711 606967 Fax +44 (0)1280 814007
www.emcourses.com

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-----Original Message-----
X-from: frank_karl-at-ardl.com [mailto:frank_karl-at-ardl.com]
Sent: 24 April 2014 15:09
To: protrain-at-emcourses.com

That is my understanding as well. In an explosion, there is not enough time
to rely upon mass transfer of oxygen to fuel the reaction. The reaction is a
rearrangement of atoms already present.

The question will be what energy threshold needs to be exceeded to start the
reaction. Low voltages and beam currents are a good approach. High
explosives may be safer because they need quite a kick from a detonator to
get started.

Of course, it is also helpful to limit the material examined so if something
goes off it won't damage the instrument.

I'd be interested if someone could address those issues.

-----Original Message-----
X-from: colijn.1-at-osu.edu [mailto:colijn.1-at-osu.edu]
Sent: Thursday, April 24, 2014 7:16 AM

Hi Stephane,

I believe that most explosives contain their own oxidizers to enable the
rapid reaction. Hence the presence of peroxide in many of the homemade
concoctions. Anyone want to clarify the issue?

Cheers,
Henk

On 4/24/2014 7:35 AM, nizets2-at-yahoo.com wrote:
}
}
} I would be happy to be corrected if I was wrong but I always thought that
an explosion was a very fast combustion.
} And that a combustion needs oxygen, which is by definition absent from a
high vacuum environment.
} Can I convince somebody with my argument?
}
} Stephane
}
} X-from: "hanke-at-mee-inc.com" {hanke-at-mee-inc.com}
} To: nizets2-at-yahoo.com
} Sent: Tuesday, April 22, 2014 4:44 PM
} Subject: [Microscopy] SEM/EDS of Gunpowder
}
}
}
}
}
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}
} Anyone ever examine/analyze gunpowder or primer samples by SEM? Any
} hints or cautions?
}
} Thanks.
}

--

Hendrik O. Colijn

*C*enter for *E*lectron *M*icroscopy & *A*nalysi*S*

1305 Kinnear Rd, Suite 100, Columbus, OH 43212

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Email: ravi.thakkar369-at-gmail.com
Name: Ravi Thakkar

Title-Subject: [Filtered] Ultrastructural pathology : Unknown object in muscle fiber

Message: Hi,

We found some electron dense material in the skeletal muscle fiber at subsarcolemmal as well as
intermyofibrillar spaces.
It can be seen in mitochondria and even in endomysium also. Higher magnification show it is not
filamentous inclusion.
Then what it could be.?

Clue : Skeletal muscle biopsy from 6 years male, Histopathology suggests Conginetal muscular dystrophy.

Images : https://www.flickr.com/photos/97321550-at-N08/13991844032/

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To Ravi:

have just had a very short view of your images on FLICKR.com (hopefully I have seen all).

My impressions so far(not to teach you!):

i) ultrastructural preservation seems to be compromised, not knowing whether due to delay in fixation, incorrect fixative [regarding mosmol, pH and / or composition] or really (true) caused by the / a disease.
ii) most, if not all mitochondria contained in the images show swelling and dysmorphic mitochondrial texture (enlarged, empty mitoch. matrix, only few remnants of cristae mitoch.) [causes: as before]
iii) most of the "dark, e-dense material" suggested to be products of mitochondrial degeneration, since usually within the former mitochondrial membrane(s).
iv) the cylindric lamellar structure in Tv32 IMHO is a degenerated mitochondrium (with a central, so called "glycogen body", said to be end-product of mitochondrial degeneration - but this never seen to be scientifically confirmed), but perhaps could be assigned also to SER/t-tubulus canal. The e-dense, round area could be {morphological equivalent} of mitochondrial energy supply (e. g./i.e. [phosphor-]lipidic material normally to be transferred into the mitochondrium via its membrane, but in the condition shown, disturbed due to mitochondrial degeneration (vice verse, means: primarily or secondarily disturbed transfer/transport mechanism).
This circumstance / possibility is more detailed and can be seen in Tv10 & Tv9, which reminds me on a (juvenile muscle biopsy) case with ATP-synthase or another enzyme deficiency.
v) if the e-dense material would be "normal" (neutral) lipids in the specimen, as seen with / after classical TEM-spec. processing, such would have been eliminated (empty vacuoles). Therefore it would be a good idea to tell us a little bit more about your routine/special processing of the biopsy.

Hope this helps anyway, unfortunately in a hurry for special reason, so I cannot comment on possible entities or similar cases/morphologies....
perhaps I will return to the thread tomorrow...

Best wishes and regards
Wolfgang

Wolfgang MUSS
EM-Lab
Univ.Inst. Pathology
SALK-LKH (Gen. Hosp.) and PMU SALZBURG
SALZBURG, AUSTRIA

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} Email: ravi.thakkar369-at-gmail.com
} Name: Ravi Thakkar
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} Title-Subject: [Filtered] Ultrastructural pathology : Unknown object in
} muscle fiber
}
} Message: Hi,
}
} We found some electron dense material in the skeletal muscle fiber at
} subsarcolemmal as well as intermyofibrillar spaces.
} It can be seen in mitochondria and even in endomysium also. Higher
} magnification show it is not filamentous inclusion.
} Then what it could be.?
}
} Clue : Skeletal muscle biopsy from 6 years male, Histopathology
} suggests Conginetal muscular dystrophy.
}
} Images : https://www.flickr.com/photos/97321550-at-N08/13991844032/
}
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So is the fear that the SEM Vacuum chamber cam blow up since the gun
powder can fuel its own combustion? So does this mean current guns and
ammo technology will still be good weapons to use in outer space?
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} werner1-at-slb.com wrote:
} }
} }
} } Modern small arms propellants (smokeless powder) are generally nitrocellulose based (there are single base and double base powders, the double base contain some nitroglycerine), granulated, and coated with deterrent coatings that encourage progressive burning - controlled deflagration - to tailor the pressure-time curve to the projectile/barrel length combination at hand.
} }
} } You can make nitrocellulose easily by nitrating cotton or paper with nitric and sulfuric acids. Note that this is something to do in controlled conditions, with proper cooling and safeguards - but my 7th grade Science Teacher, Miss Wyle, showed us and actually may be the person ultimately responsible for the course my life has taken (she was great).
} }
} }
}
} Right, sorry, I was thinking about black powder, not modern
} smokeless powder which is what "gunpowder" currently means.
} Andrew was typing his more detailed explanations while I was
} still sending mine.
}
} This is a bit OT, but:
}
}
} http://explosives.mst.edu/media/academic/explosives/documents/camp/2014ExplosivesCamp.pdf
}
} if you have a kid 16 or older (or if you're young enough to pass
} for a high-school kid, which definitely excludes me), check this out!
}
} Rick Mott (guilty of making NI3 and other noisy fun stuff as
} a teenager)
}
} ==============================Original Headers==============================
} 5, 18 -- From rmott-at-pulsetor.com Thu Apr 24 10:05:42 2014
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} 5, 18 -- To: microscopy-at-microscopy.com
} 5, 18 -- Subject: Re: [Microscopy] Fw: SEM/EDS of Gunpowder
} 5, 18 -- References: {201404241445.s3OEjBij000870-at-ns.microscopy.com}
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2, 32 -- From mmilewski-at-comcast.net Thu Apr 24 15:10:17 2014
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I think the concern is that the energetic material could potentially decompose into it's reaction products, liberating some heat and gas - CO2, H2O, N2, CO - that might be a pain for the vacuum system to deal with.

But yes, current guns and ammunition will work in vacuum. You would need less volatile lubricants on moving parts, or solid-state anti-friction coatings (hard chrome or spray moly might work), and the barrel might be subject to excessive copper fouling - the bullet jacket riding directly on the lands might gall - but we moly coat match bullets now, so I can't immediately think of any insurmountable difficulties. Except - you need a good backstop (here on Earth a substantial dirt berm works well), and in the absence of a significant gravitational field, recoil will send the shooter tumbling / spinning. And - with no wind to read and compensate, where is the challenge?

Andrew

-----Original Message-----
X-from: mmilewski-at-comcast.net [mailto:mmilewski-at-comcast.net]
Sent: Thursday, April 24, 2014 3:21 PM
To: Andrew Werner

So is the fear that the SEM Vacuum chamber cam blow up since the gun
powder can fuel its own combustion? So does this mean current guns and
ammo technology will still be good weapons to use in outer space?
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
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} ----------------------------------------------------------------------------
}
} werner1-at-slb.com wrote:
} }
} }
} } Modern small arms propellants (smokeless powder) are generally nitrocellulose based (there are single base and double base powders, the double base contain some nitroglycerine), granulated, and coated with deterrent coatings that encourage progressive burning - controlled deflagration - to tailor the pressure-time curve to the projectile/barrel length combination at hand.
} }
} } You can make nitrocellulose easily by nitrating cotton or paper with nitric and sulfuric acids. Note that this is something to do in controlled conditions, with proper cooling and safeguards - but my 7th grade Science Teacher, Miss Wyle, showed us and actually may be the person ultimately responsible for the course my life has taken (she was great).
} }
} }
}
} Right, sorry, I was thinking about black powder, not modern
} smokeless powder which is what "gunpowder" currently means.
} Andrew was typing his more detailed explanations while I was
} still sending mine.
}
} This is a bit OT, but:
}
}
} http://explosives.mst.edu/media/academic/explosives/documents/camp/2014ExplosivesCamp.pdf
}
} if you have a kid 16 or older (or if you're young enough to pass
} for a high-school kid, which definitely excludes me), check this out!
}
} Rick Mott (guilty of making NI3 and other noisy fun stuff as
} a teenager)
}
} ==============================Original Headers==============================
} 5, 18 -- From rmott-at-pulsetor.com Thu Apr 24 10:05:42 2014
} 5, 18 -- Received: from p3plsmtpa08-05.prod.phx3.secureserver.net (p3plsmtpa08-05.prod.phx3.secureserver.net [173.201.193.106])
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} 5, 18 -- Date: Thu, 24 Apr 2014 11:05:26 -0400
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} 5, 18 -- To: microscopy-at-microscopy.com
} 5, 18 -- Subject: Re: [Microscopy] Fw: SEM/EDS of Gunpowder
} 5, 18 -- References: {201404241445.s3OEjBij000870-at-ns.microscopy.com}
} 5, 18 -- In-Reply-To: {201404241445.s3OEjBij000870-at-ns.microscopy.com}
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2, 32 -- Subject: Re: [Microscopy] Re: Fw: SEM/EDS of Gunpowder
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12, 35 -- From werner1-at-slb.com Thu Apr 24 15:38:15 2014
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12, 35 -- From: Andrew Werner {werner1-at-slb.com}
12, 35 -- To: "mmilewski-at-comcast.net" {mmilewski-at-comcast.net}
12, 35 -- CC: "microscopy-at-microscopy.com" {microscopy-at-microscopy.com}
12, 35 -- Subject: RE: [Microscopy] Fw: SEM/EDS of Gunpowder
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Boy has this gotten off-topic, but fun nonetheless!

I would say that the reactions (deflagration?) will still occur, so ammo
should work in outer space. However, you may want to remember Newton's
3rd Law! Also the combustion products will significantly affect the
local vacuum.

Cheers,
Henk

On 4/24/2014 4:12 PM, mmilewski-at-comcast.net wrote:
}
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} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
} To Subscribe/Unsubscribe --http://www.microscopy.com/MicroscopyListserver
} On-Line Helphttp://www.microscopy.com/MicroscopyListserver/FAQ.html
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}
} So is the fear that the SEM Vacuum chamber cam blow up since the gun
} powder can fuel its own combustion? So does this mean current guns and
} ammo technology will still be good weapons to use in outer space?
} } ----------------------------------------------------------------------------
} } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
} } To Subscribe/Unsubscribe --http://www.microscopy.com/MicroscopyListserver
} } On-Line Helphttp://www.microscopy.com/MicroscopyListserver/FAQ.html
} } ----------------------------------------------------------------------------
} }
} } werner1-at-slb.com wrote:
} } }
} } }
} } } Modern small arms propellants (smokeless powder) are generally nitrocellulose based (there are single base and double base powders, the double base contain some nitroglycerine), granulated, and coated with deterrent coatings that encourage progressive burning - controlled deflagration - to tailor the pressure-time curve to the projectile/barrel length combination at hand.
} } }
} } } You can make nitrocellulose easily by nitrating cotton or paper with nitric and sulfuric acids. Note that this is something to do in controlled conditions, with proper cooling and safeguards - but my 7th grade Science Teacher, Miss Wyle, showed us and actually may be the person ultimately responsible for the course my life has taken (she was great).
} } }
} } }
} }
} } Right, sorry, I was thinking about black powder, not modern
} } smokeless powder which is what "gunpowder" currently means.
} } Andrew was typing his more detailed explanations while I was
} } still sending mine.
} }
} } This is a bit OT, but:
} }
} }
} } http://explosives.mst.edu/media/academic/explosives/documents/camp/2014ExplosivesCamp.pdf
} }
} } if you have a kid 16 or older (or if you're young enough to pass
} } for a high-school kid, which definitely excludes me), check this out!
} }
} } Rick Mott (guilty of making NI3 and other noisy fun stuff as
} } a teenager)
} }
} } ==============================Original Headers==============================
} } 5, 18 -- Fromrmott-at-pulsetor.com Thu Apr 24 10:05:42 2014
} } 5, 18 -- Received: from p3plsmtpa08-05.prod.phx3.secureserver.net (p3plsmtpa08-05.prod.phx3.secureserver.net [173.201.193.106])
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Email: matthew.hannah-at-phe.gov.uk
Name: Matthew Hannah

Organization: Public Health England

Title-Subject: [Filtered] embedding Cytodex3 beads

Message: Hi, has anyone any experience of embedding Cytodex3 (or other dextran beads) in plastic for
ultrathin sectioning and TEM? I have someone growing (polarized) cells on them and would like to
have a look at the ultrastructure. I have tried a number of resins and protocols (although nothing
longer than two days so far) but the plastic does not appear to infiltrate the dextran. Any tricks
or tips gratefully received. Thanks

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Title-Subject: [Filtered] 3rd Annual OHSU-FEI Bioimaging at the Nanoscale Symposium

Message: Dear EM community,
FEI Company and the Oregon Health and Sciences University (OHSU) are pleased to announce that
registration is now open for the Bioimaging at the Nanoscale Symposium. The symposium will be hosted
by the OHSU Center for Spatial Systems Biology and the FEI-OHSU Living Lab for Cell Biology, in
Portland, Oregon, June 4th-6th, 2014. This symposium will showcase cutting edge microscopy
technologies for imaging biological systems at multiple resolution scales. Presentations will cover
several imaging modalities that include molecular electron microscopy (EM), super-resolution
microscopy (SRM), and correlative light and electron microscopy (CLEM). World-renowned experts from
these fields will discuss the most recent developments in imaging technologies and how the new tools
have helped transform our understanding of biology.
On June 4, FEI will host a full day of multiple workshops utilizing the suite of FEI microscopes
currently installed at OHSU (Titan Krios, Tecnai with iCorr, Helios NanoLab 650). These workshops,
run by FEI and OHSU personnel, will give microscopists and non-microscopists an opportunity to learn
about current and cutting edge applications from FEI.
Attendees are strongly encouraged to both learn from FEI and share your experiences through poster
and platform presentations.
Registration is free and includes complimentary breakfast, lunch and coffee for all participants.
Invited speakers include:
Â• Abraham Koster (Leiden U.)
Â• Gregor Heiss (FEI Co.)
Â• Robert Campbell (University of Alberta)
Â• James Jeffers (Intel Co.)
Â• Michael Chapman (OHSU)
Â• Catherine Galbraith (OHSU)
Â• James Galbraith (OHSU)
Â• Andrew Ward (Scripps)
Â• Gabe Lander (Scripps)
Â• Adam Frost (U. of Utah)
Â• Missy Wong (OHSU)
Â• Doug Keene (Shriners Hospital for Children)
Â• Danielle Jorgens (OHSU)
Â• Anke Mulder (OHSU)
Â• Gregory Scott (OHSU)
Â• Claudia Lopez (MMC -at- OHSU)
Â• Stefanie Kaech Petrie (ALMC -at- OHSU)

For registration click here:
http://marketing.fei.com/acton/fs/blocks/showLandingPage/a/4019/p/p-012d/t/page/fm/0

Site address:
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From discounted_meds8-at-ne.jp Sat Apr 26 03:49:14 2014
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Hi everyone,

I was wondering whether it is safe to section 400 nm thick section with ultra diamond knife, or whether I have to use a histo knife

thank you

John

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Hi John,
We routinely use old Ultra diamond knives to cut 1 um thick sections. But I must say I love my Histo diamond knife. If you are cutting a lot of thick sections a Histo diamond knife equals true happiness and I encourage you to buy one if you can afford it - 6mm will rock your world;-)
best,
Beth

==============================Original Headers==============================
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2, 21 -- From: "Beth Richardson" {beth-at-plantbio.uga.edu}
2, 21 -- Subject: [Microscopy] 400 nm sections -what kind of knife
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I have to agree with Beth. I used to use older ultra knives and live with
some scratches, but someone brought in a 7 mm Histo knife that I made fun
of ... and now I wouldn't part with it for anything. I can cut 1
micrometer sections on it and then, *without moving it or changing
anything else*, silver sections.

Aloha,
Tina

} I was wondering whether it is safe to section 400 nm thick section with ultra diamond knife, or whether I have to use a histo knife

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

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From drugstore-cheapest11-at-prudentialdoss.com Sun Apr 27 03:14:13 2014
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Dear John,
agree too with Tina & Beth, also with using "older ultra knives" prior to "resharpening".
A separate {HISTO} -knife or special diamond dedicated for semithin sectioning would add some personal joy and happiness, I guess.
at least one {Histo} knife (6-7 mm sectioning edge) should be standard in the prep./cutting-armamentarium when doing sectioning resin blocks for LM prior to TEM.
It certainly will pay off.

Best wishes and good luck,
Wolfgang

Wolfgang MUSS
EM-Lab
Univ. Inst. Pathology
SALK-LKH (Gen. Hospital) &
PMU (Paracelsus Med. Univ.) SALZBURG
Austria

} -----Ursprüngliche Nachricht-----
} Von: tina-at-pbrc.hawaii.edu [mailto:tina-at-pbrc.hawaii.edu]
} Gesendet: Samstag, 26. April 2014 22:45
} An: Muß Wolfgang
} Betreff: [Microscopy] Re: 400 nm sections -what kind of knife
}
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} I have to agree with Beth. I used to use older ultra knives and live
} with some scratches, but someone brought in a 7 mm Histo knife that I
} made fun
} of ... and now I wouldn't part with it for anything. I can cut 1
} micrometer sections on it and then, *without moving it or changing
} anything else*, silver sections.
}
} Aloha,
} Tina
}
} } I was wondering whether it is safe to section 400 nm thick section
} with ultra diamond knife, or whether I have to use a histo knife
}
} ***********************************************************************
} *****
} * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu
} *
} * Biological Electron Microscope Facility * (808) 956-6251
} *
} * University of Hawaii at Manoa *
} http://www.pbrc.hawaii.edu/bemf*
} ***********************************************************************
} *****
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} Re by Beth Richardson: beth-at-plantbio.uga.edu Sa 26.04.2014 18:02
}
} Hi John,
} We routinely use old Ultra diamond knives to cut 1µm thick sections.
} But I must say I love my Histo diamond knife. If you are cutting a lot
} of thick sections a Histo diamond knife equals true happiness and I
} encourage you to buy one if you can afford it - 6mm will rock your
} world;-)
} best,
} Beth
}
}
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Dear friends,
Iâ€™m preparing a poster (A1 size) on transmission electron microscopy (TEM) topic to decorate our characterisation centre. We have two TEMs a Jeol 2000FX and a FEI Tecnai F20 along with typical optical microscopes and SEMs, FIB plus surface analysis equipment. Our major interests are materials and physical sciences teaching and research. My manager suggested some entry level information in the poster for public visitors including 16yrs old and their parents. I used to go to conferences with research posters. It wasnâ€™t as difficult as this.
Can you give me some inspirations please? Any suggestions, text or websites etc are all appreciated.
Best regards,
Zhou

Dr Zhaoxia Zhou
Experimental Officer
Loughborough Materials Characterisation Centre (LMCC)
Department of Materials
Loughborough University
Leicestershire
LE11 3TU

Tel: 01509 223163
Fax: 01509 234225

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Hello, Ravi,

I agree with Wolfgang that your material has been poorly fixed, which complicates the interpretation of your images. The material in question appears to be coagulated Z-band material, which is extremely electron-dense fibers that sometimes appear in disrupted myocytes. I used to study muscular dystrophy in myotonic mice, and would come across myocytes with disrupted Z-band material, some of which had the coagulation patches which you are showing.

Ed Haller

Edward Haller, Lab Manager
University of South Florida
Department of Integrative Biology
Electron Microscopy Core
SCA 110
4202 East Fowler Avenue
Tampa, FL 33620
(813)974-2676
ehaller-at-usf.edu
Office: ISA 1046
http://biology.usf.edu/ib/research/facilities/

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Email: ravi.thakkar369-at-gmail.com
Name: Ravi Thakkar

Title-Subject: [Filtered] Ultrastructural pathology : Unknown object in muscle fiber

Message: Hi,

We found some electron dense material in the skeletal muscle fiber at subsarcolemmal as well as
intermyofibrillar spaces.
It can be seen in mitochondria and even in endomysium also. Higher magnification show it is not
filamentous inclusion.
Then what it could be.?

Clue : Skeletal muscle biopsy from 6 years male, Histopathology suggests Conginetal muscular dystrophy.

Images : https://www.flickr.com/photos/97321550-at-N08/13991844032/

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Cryo-EM Specialist - YALE UNIVERSITY SCHOOL OF MEDICINE

A Cryo-EM specialist is sought by the Center for Cell and Molecular
Imaging (CCMI) at Yale University School of Medicine. CCMI provides
faculty members, postdoctoral fellows, and students with access to
instrumentation and expertise for cryoelectron microscopy and other
imaging technologies. The successful candidate should have a PhD degree in
biology, physics, engineering, or a related field, and should be
experienced in cryo-EM specimen preparation and image acquisition for
single particles and electron tomography. He/she will acquire data and
assist users in the wide variety of projects that are carried out in the
highly collaborative environment of Yale©ös Cryo-EM facility. He/she will
also seek to optimize the instruments and procedures for data collection.
The Yale facility has two Tecnai F20 electron microscopes, one of which is
equipped with a K2 electron-counting camera, as well as a T12 microscope,
Vitrobot, and other sample-preparation devices. Applications, including a
cover letter, curriculum vitae, and the names of three referees, should
be sent to Dr. Xinran Liu, Director of the EM Core Facility at Yale School
of Medicine (xinran.liu-at-yale.edu) and are due by June 1, 2014.
Applications from women and minority scientists are encouraged. Yale is an
Affirmative Action/Equal Opportunity Employer.

Center for Cellular and Molecular Imaging
http://medicine.yale.edu/ccmi/em

==============================Original Headers==============================
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Hi Zhaoxia,

One metric I use when communicating to laymen is "the thickness of the
human hair" which is about 100 um. Everyone has a feeling for how thick a
human hair is, but when you say nanometer they don't have a clue about
that. So if you have a 100 nm particle, you say, "If a human hair were as
thick as this room, then this object would be about 1 centimeter across!"
(Well, OK, I say half an inch because the US is still in the SI
backwater...)

For TEMs and SEMs, I make the analogy to an optical microscope (which a
some people have experience with) or a magnifying glass, if that fails.
The reason why we use electrons is because they are "smaller than light"
and they let us see "different things inside."

Finally, the most important thing: pretty pictures. Be sure to have RGB
pictures of something dazzling that makes them want to pore over it and see
what that is. Ideally, it should have some parallels with objects people
see in the big world. For example, I once used three plasmon images of a
FIB lamella to make an RGB which looked like stained glass. The scientific
content was pretty low, but people loved looking at the picture and it got
them interested. I also have a picture of a sample prepared for AFM that
looks like the Sauron's tower from Lord of the Rings -- I'll probably use
that soon. I find that what you and I consider pretty pictures are not
always the same as what a layman considers pretty. After all, they haven't
spent the last N years looking at microscopic objects.

Hope this helps!

Zack Gainsforth
Space Sciences Laboratory, UC Berkeley
7 Gauss Way
Berkeley, CA 94720
cell: 626-437-9186 |--//cell: 626-437-9186--|
zackg-at-ssl.berkeley.edu

On Mon, Apr 28, 2014 at 4:13 AM, |--Z.Zhou-at-lboro.ac.uk--| wrote:

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--| Dear friends,
--| I'm preparing a poster (A1 size) on transmission electron microscopy (TEM)
--| topic to decorate our characterisation centre. We have two TEMs a Jeol
--| 2000FX and a FEI Tecnai F20 along with typical optical microscopes and
--| SEMs, FIB plus surface analysis equipment. Our major interests are
--| materials and physical sciences teaching and research. My manager suggested
--| some entry level information in the poster for public visitors including
--| 16yrs old and their parents. I used to go to conferences with research
--| posters. It wasn't as difficult as this.
--| Can you give me some inspirations please? Any suggestions, text or
--| websites etc are all appreciated.
--| Best regards,
--| Zhou
--|
--| Dr Zhaoxia Zhou
--| Experimental Officer
--| Loughborough Materials Characterisation Centre (LMCC)
--| Department of Materials
--| Loughborough University
--| Leicestershire
--| LE11 3TU
--|
--| Tel: 01509 223163
--| Fax: 01509 234225

==============================Original Headers==============================
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Hi Zhou

I did this once years ago. We decided that one point we wanted to get across was the amazing scale of magnification provided by TEM.
We found that people, especially the general public (and I also) are astounded by what high magnifications actually mean in real terms. We routinely deal with numbers like 1000x, and 150,000x without thinking. Also what does nanometre mean? We simply did some back of the envelope calculations. Then we could say things like.

We had a picture of the earth. If the earth was 1 mm diameter then the building we are in would be 1 nm long.

If we were looking at really high magnification at some sample at 500,000x (possible with the F20?) then it is equivalent to orbiting the earth and only just being able to read the text on this poster.

Our lab works on wool. So we had a micrograph of some intermediate filaments (7.6 nm diameter) and then indicated if the filament pictured on the poster was real size then the wool fibre containing it would be 640 m across.

I am sure there are other samples where a similar exercise can be done. Once people "get" what scale really means then it is easier to wow them with the technical aspects of how you manage this miracle of magnification.

Good luck
Duane

AgResearch Limited | Lincoln Research Centre | New Zealand
Dr Duane P Harland, Senior Scientist
T +64 3 321 8710Â E duane.harland-at-agresearch.co.nz

-----Original Message-----
X-from: Z.Zhou-at-lboro.ac.uk [mailto:Z.Zhou-at-lboro.ac.uk]
Sent: Monday, 28 April 2014 11:16 p.m.
To: Harland, Duane

Dear friends,
Iâ€™m preparing a poster (A1 size) on transmission electron microscopy (TEM) topic to decorate our characterisation centre. We have two TEMs a Jeol 2000FX and a FEI Tecnai F20 along with typical optical microscopes and SEMs, FIB plus surface analysis equipment. Our major interests are materials and physical sciences teaching and research. My manager suggested some entry level information in the poster for public visitors including 16yrs old and their parents. I used to go to conferences with research posters. It wasnâ€™t as difficult as this.
Can you give me some inspirations please? Any suggestions, text or websites etc are all appreciated.
Best regards,
Zhou

Dr Zhaoxia Zhou
Experimental Officer
Loughborough Materials Characterisation Centre (LMCC) Department of Materials Loughborough University Leicestershire
LE11 3TU

Tel: 01509 223163
Fax: 01509 234225

==============================Original Headers==============================
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Email: mikroskop-at-gmail.com
Name: Jack Hietpas

Title-Subject: [Filtered] ultramicrotome section preparation for SEM

Message: Hello Microscopists/ Microtomists - I am currently making ultramicrotome sections
(500nm-1um, cross- and longitudinal-sections) of individual hair samples that are embedded in
Embed-812. I am interested in high magnification imaging of the hair structure using SEM which
should be simple enough.... however the sections are constantly folding and wrinkling once they are
transferred and dried onto a sample support for SEM examination. The wrinkling usually negates
proper examination. I was wondering if there are any tricks-of-the-trade that could over come this
issue?
Below is my current method:

1. Cut sections with ultramicrotome

2. Transfer sections (using an eyelash probe) to a gold-coated microscope coverglass (functioning as
a sample support) that contains a micro-drop of water.

3. While the section is still floating on the water micro-drop, it is flattened using chloroform
vapor. This works very well.

4. Either allow the water micro-drop to evaporate or gently wick the water away using a pointed
piece of filter paper. IT IS HERE WHERE THE SECTIONS GET WRINKLED!

5. Gold-coat and examine using SEM.

Any suggestions on how to remove the water and have the thin section lay flat on the coverglass
surface would be greatly appreciated. Perhaps a completely new substrate would work better! Is
this an issue when using a TEM grid?

Thanks for your time and any information you can provide,

-Jack

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Having worked almost exclusively with tungsten SEMs, the example I have
given for years is that taking 4x5 Polaroids at 100,000x would require a
line of photos lined up for 100m (longer than a US football field) to show
the area between two lines on a ruler that are 1mm apart. Even most
Americans are at least somewhat familiar with a mm (about 1/32") and if they
weren't, I'd show them my ruler.

The TEM (especially the new ones) are at least an order of magnitude better,
so at a million X, the mm lines are a kilometer apart (.8 miles in the US).

Ken Converse
owner

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

-----Original Message-----
X-from: Duane.Harland-at-agresearch.co.nz [mailto:Duane.Harland-at-agresearch.co.nz]

Sent: Monday, April 28, 2014 5:23 PM
To: kenconverse-at-qualityimages.biz

Hi Zhou

I did this once years ago. We decided that one point we wanted to get across
was the amazing scale of magnification provided by TEM.
We found that people, especially the general public (and I also) are
astounded by what high magnifications actually mean in real terms. We
routinely deal with numbers like 1000x, and 150,000x without thinking. Also
what does nanometre mean? We simply did some back of the envelope
calculations. Then we could say things like.

We had a picture of the earth. If the earth was 1 mm diameter then the
building we are in would be 1 nm long.

If we were looking at really high magnification at some sample at 500,000x
(possible with the F20?) then it is equivalent to orbiting the earth and
only just being able to read the text on this poster.

Our lab works on wool. So we had a micrograph of some intermediate filaments
(7.6 nm diameter) and then indicated if the filament pictured on the poster
was real size then the wool fibre containing it would be 640 m across.

I am sure there are other samples where a similar exercise can be done. Once
people "get" what scale really means then it is easier to wow them with the
technical aspects of how you manage this miracle of magnification.

Good luck
Duane

AgResearch Limited | Lincoln Research Centre | New Zealand
Dr Duane P Harland, Senior Scientist
T +64 3 321 8710Â E duane.harland-at-agresearch.co.nz

-----Original Message-----
X-from: Z.Zhou-at-lboro.ac.uk [mailto:Z.Zhou-at-lboro.ac.uk]
Sent: Monday, 28 April 2014 11:16 p.m.
To: Harland, Duane

Dear friends,
Iâ€™m preparing a poster (A1 size) on transmission electron microscopy (TEM)
topic to decorate our characterisation centre. We have two TEMs a Jeol
2000FX and a FEI Tecnai F20 along with typical optical microscopes and SEMs,
FIB plus surface analysis equipment. Our major interests are materials and
physical sciences teaching and research. My manager suggested some entry
level information in the poster for public visitors including 16yrs old and
their parents. I used to go to conferences with research posters. It
wasnâ€™t as difficult as this.
Can you give me some inspirations please? Any suggestions, text or websites
etc are all appreciated.
Best regards,
Zhou

Dr Zhaoxia Zhou
Experimental Officer
Loughborough Materials Characterisation Centre (LMCC) Department of
Materials Loughborough University Leicestershire
LE11 3TU

Tel: 01509 223163
Fax: 01509 234225

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Hi Jack

Try without water and colllect section gently with adhesive tape and good
luck!

yorgos

Dr Yorgos Nikas
Athens Innovative Microscopy
Skra 36 Voula 16673 GREECE

Tel/fax +30 210 8957677
mobile +30 6945 107477
www.eikonika.net www.aim.cat
************************************

----- Original Message -----
X-from: {microscopylistserver-noreply-at-microscopy.com}
To: {eikonika-at-otenet.gr}
Sent: Tuesday, April 29, 2014 1:09 AM

Ian,
thanks for your help. It had been on the point... ;-)

Here is another link a found after searching for "broad mite":
http://www.google.de/imgres?imgurl=http%3A%2F%2Fwww.sel.barc.usda.gov%2Facari%2Fimages%2Fbroad%2Fegg.jpg&imgrefurl=http%3A%2F%2Fwww.sel.barc.usda.gov%2Facari%2Fcontent%2Fbroad%2Fa.html&h=331&w=419&tbnid=aqG3xtaawrHITM%3A&zoom=1&docid=lvTbzUBHnO6QkM&ei=TYBfU-uUMYeHtAaw14HwBg&tbm=isch&client=firefox-a&iact=rc&uact=3&dur=547&page=1&start=0&ndsp=67&ved=0CHwQrQMwDA

Best wishes,
Stefan

-----------------------------------------------------
Stefan Diller - Scientific Photography
Arndtstrasse 22
D - 97072 Wuerzburg Germany
++49-931-7848700 Phone
++49-931-7848701 Fax
++49-175-7177051 Mobile

Websites:
www.nanoflight.info
www.stefan-diller.com
www.electronmicroscopy.info
www.elektronenmikroskopie.info
www.zwillingsprojekt.de
www.assisi.de
Anfahrt: http://Mail.map24.com/Stefan.Diller
-----------------------------------------------------

Am 29.04.14 06:22, schrieb Ian Hallett:
} Stefan
} You are almost right the object is the egg of a mite (possibly from images we have taken a broad mite).
} Ian
} Ian Hallett
} Plant and Food Research
} Auckland New Zealand
}
} On Fri, Apr 18, 2014 at 6:33 AM, {stefan.diller-at-t-online.de {mailto:stefan.diller-at-t-online.de} } wrote:
}
}
}
}
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
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} On-Line {http://www.microscopy.com/MicroscopyListserverOn-Line} Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
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}
} Dear All,
} maybe there is someone out there before going into Easter holidays to share a bit of knowledge.
} I just founds a strange detail on a specimen (the upper leaf surface of a cucumber plant):
} See images here:
} www.electronmicroscopy.info/customers/cucumber/index.htm {http://www.electronmicroscopy.info/customers/cucumber/index.htm}
} There is also a PDF with EDS data, but it is not very specific since I put a whole lot of Platinum on the surface because of a
} nanoflight movie I did with this surface:
} www.electronmicroscopy.info/customers/cucumber/cucumber_eds.pdf
} {http://www.electronmicroscopy.info/customers/cucumber/cucumber_eds.pdf}
} A little bit of Sulfur and Ca is showing up, with Carbon.
} Any idea what this might be?
}
} Best wishes, happy Easter,
} Stefan
}
} --
}
}
} -----------------------------------------------------
} Stefan Diller - Scientific Photography
} Arndtstrasse 22
} D - 97072 Wuerzburg Germany
} ++49-931-7848700 {tel:%2B%2B49-931-7848700} Phone
} ++49-931-7848701 {tel:%2B%2B49-931-7848701} Fax
} ++49-175-7177051 {tel:%2B%2B49-175-7177051} Mobile
}
} Websites:
} www.nanoflight.info {http://www.nanoflight.info/}
} www.stefan-diller.com {http://www.stefan-diller.com/}
} www.electronmicroscopy.info {http://www.electronmicroscopy.info/}
} www.elektronenmikroskopie.info {http://www.elektronenmikroskopie.info/}
} www.zwillingsprojekt.de {http://www.zwillingsprojekt.de/}
} www.assisi.de {http://www.assisi.de/}
} Anfahrt: http://Mail.map24.com/Stefan.Diller {http://mail.map24.com/Stefan.Diller}
} -----------------------------------------------------
}
} ==============================Original Headers==============================
} 6, 20 -- From stefan.diller-at-t-online.de {mailto:stefan.diller-at-t-online.de} Thu Apr 17 13:22:15 2014
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8, 22 -- From stefan.diller-at-t-online.de Tue Apr 29 05:40:06 2014
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Have you tried warming the slide on a hot plate or slide warmer? One
warm enough to keep the sections expanded (or to expand them with heat
instead of chloroform) - what one would use to warm the sections when
doing toluidine blue staining. 60 to 90 deg C.

Phil

} Email: mikroskop-at-gmail.com
} Name: Jack Hietpas
}
} Title-Subject: [Filtered] ultramicrotome section preparation for SEM
}
} Message: Hello Microscopists/ Microtomists - I am currently making ultramicrotome sections
} (500nm-1um, cross- and longitudinal-sections) of individual hair samples that are embedded in
} Embed-812. I am interested in high magnification imaging of the hair structure using SEM which
} should be simple enough.... however the sections are constantly folding and wrinkling once they are
} transferred and dried onto a sample support for SEM examination. The wrinkling usually negates
} proper examination. I was wondering if there are any tricks-of-the-trade that could over come this
} issue?
} Below is my current method:
}
} 1. Cut sections with ultramicrotome
}
} 2. Transfer sections (using an eyelash probe) to a gold-coated microscope coverglass (functioning as
} a sample support) that contains a micro-drop of water.
}
} 3. While the section is still floating on the water micro-drop, it is flattened using chloroform
} vapor. This works very well.
}
} 4. Either allow the water micro-drop to evaporate or gently wick the water away using a pointed
} piece of filter paper. IT IS HERE WHERE THE SECTIONS GET WRINKLED!
}
} 5. Gold-coat and examine using SEM.
}
} Any suggestions on how to remove the water and have the thin section lay flat on the coverglass
} surface would be greatly appreciated. Perhaps a completely new substrate would work better! Is
} this an issue when using a TEM grid?
}
} Thanks for your time and any information you can provide,
}
} -Jack
}
} Login Host: 63.167.71.243
} Listserver Email Form V - 20120416
} ---------------------------------------------------------------------------
}
}
}
}

--
Philip Oshel
Microscopy Facility Supervisor
Biology Department
024C Brooks Hall
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576

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Just in case we haven't hammered the topic "can explosives and gunpowder work in a vacuum" into scrap metal, here's a link to a Wired article that covers a Mythbusters shooting in a vacuum.

http://www.wired.com/2014/02/hidden-physics-mythbusters-bullet-baloney/

It's at the end of the article, but the recoil on the bent rifle barrel was interesting too!

Stay safe...........
Frank

____________

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Hi, Jack,

First of all, I would not coat the coverslip until after drying the sections. Second, you might try a technique posted by another researcher on the listserver a few years ago:

"I cut1Âµ sections (and 5Âµ sections) with Diatome diamond knives mostly samples embedded in Polybed 812. I would transfer them onto a glass slide with a wooden stick or a loop. I would put the slide on a hotplate 60ËšC and then invert a large glass Petri dish over the slide. I would place a cotton swab dipped in acetone under the dish with the slide. The acetone vapor + the heat would flatten the sections and as the drop of water evaporated the sections would anneal to the slide. It worked great for me."

Good luck
Dean Abel
Biological Sciences University of Iowa, Iowa City IA USA

I tried Dean's technique, and it works with some of my samples. What I usually do is to work with glass slides, put a 3/4 to 1 inch diameter drop of water on them with my sections on the water, and use a flame from a Bunsen burner to dry the drop of water down, passing the slide in and out of the top of the flame (I use a Touch-O-Matic burner, which gives me a small, controlled flame), and holding the slide at about a 30 degree angle so that the water dries out from under the sections. You must be careful with this technique to not get the water so hot that it boils, though!. Usually, if the technique is done right, and your slides are clean and your water is clean, the sections will tend to go to the top edge of the drop of water (uphill side of the slide), and the water will dry out from under the sections, leaving the sections perfectly flat. You need to play around with the size of your flame and how hot you get your slide, but once you get your conditions right, this technique is very consistent. After all of the water is dried off the slide, I pass the slide above the flame for a few more seconds to firmly adhere the sections to the slide before allowing the slide to cool, and before staining (if doing toluidine blue staining). In your case, once the sections are flat, you can coat them, and they are ready for the SEM. Take a diamond scribe and score your slide, break it small enough for your SEM stage, coat your sample, and you're ready for viewing.

Ed

Edward Haller, Lab Manager
University of South Florida
Department of Integrative Biology
Electron Microscopy Core
SCA 110
4202 East Fowler Avenue
Tampa, FL 33620
(813)974-2676
ehaller-at-usf.edu
Office: ISA 1046
http://biology.usf.edu/ib/research/facilities/

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Email: mikroskop-at-gmail.com
Name: Jack Hietpas

Title-Subject: [Filtered] ultramicrotome section preparation for SEM

Message: Hello Microscopists/ Microtomists - I am currently making ultramicrotome sections
(500nm-1um, cross- and longitudinal-sections) of individual hair samples that are embedded in
Embed-812. I am interested in high magnification imaging of the hair structure using SEM which
should be simple enough.... however the sections are constantly folding and wrinkling once they are
transferred and dried onto a sample support for SEM examination. The wrinkling usually negates
proper examination. I was wondering if there are any tricks-of-the-trade that could over come this
issue?
Below is my current method:

1. Cut sections with ultramicrotome

2. Transfer sections (using an eyelash probe) to a gold-coated microscope coverglass (functioning as
a sample support) that contains a micro-drop of water.

3. While the section is still floating on the water micro-drop, it is flattened using chloroform
vapor. This works very well.

4. Either allow the water micro-drop to evaporate or gently wick the water away using a pointed
piece of filter paper. IT IS HERE WHERE THE SECTIONS GET WRINKLED!

5. Gold-coat and examine using SEM.

Any suggestions on how to remove the water and have the thin section lay flat on the coverglass
surface would be greatly appreciated. Perhaps a completely new substrate would work better! Is
this an issue when using a TEM grid?

Thanks for your time and any information you can provide,

-Jack

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Title-Subject: [Filtered] FIB Technician Position in Applied Materials Company

Message: Our TEM/FIB lab is located in Portland, Oregon and is equipped with a state-of-the-art FEI
Osiris TEM and a Helios 600 FIB/SEM. FIB technician position will focus on TEM sample preparation
of MRAM and FinFET multi-layer films by hands-on operation of ex-situ or in-situ lift-out systems as
needed. Successful candidate will utilize their SEM/FIB experience to locate and successfully catch
sub-micron features (as small as {100 nm) with } 90% success rate for cross-sectional TEM observations.

Term:
Must be available for a minimum of 6 months. Term could be extended to 12 months. It is highly
possible to turn into a regular full time position for exceptional candidates.

Work Schedule:
Requires flexibility to accommodate customersÂ’ urgent TEM/FIB requests on swing shifts.

Education:
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Science, Chemistry, or Physics, etc.

Experience:
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prep techniques. Hands-on experience on manual TEM sample preparation a plus.

Skills:
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procedures.
Must be able to work in a fast paced, multi-tasking environment where priorities may change often.
Capable of utilizing time management skills and meeting expected completion dates.
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Title-Subject: [Filtered] Richardson's Stain for Plastic Sections

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Title-Subject: [Filtered] Slit sense organ

Message: Are there any cross sectional images of slit sense organs as described in

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Hi, Vickie-

I use Richardson's all the time. Try making up fresh components:

1% methylene blue in 1% sodium borate
and
1% Azure II in distilled water

I keep the components in dark brown bottles for several years. When I need
some, I mix them 1:1 (5 ml each) and suck the stain up into a 10ml syringe
fitted with a 0.45 or 0.2 micrometer filter. This way, when you stain
slides it's always freshly filtered. I stain for 40 sec at 60C. It seems
to keep OK in the syringe for weeks to months, and the individual
components in the brown bottles for years.

Aloha,
Tina

} Message: I am trying to stain plastic sections (LR White, Lowicryl and Embed 812 with Richardson's
} stain (1960's)and I have precipitate in the solution.
}
} The precipitates are too large for filtration. Does anyone know what will solubilize them and allow
} for the tissues/cells to take up stain within these plastic ultrathin sections?
}
} I look on the light microscope to see what kinds of cells I have as a preliminary region for grids
} on the TEM.
}
} Thanks!
} Vickie
}
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****************************************************************************
* Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu *
* Biological Electron Microscope Facility * (808) 956-6251 *
* University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf*
****************************************************************************

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Good morning all,
thanks Tina, for your really appreciated reply with the practical statements,

dear Vickie,

for me (I am using a "modified Richardson's", following the recipe of Humphrey and Pittman 1974 = AMbF*-stain for semithin plastic sections) the question primarily is:
why you have {precipitates - too large for filtration - in your solution} ?.

I haven't had any disturbing precipitates in my staining solutions (Toluidine blue, Richardson's, Humphrey and Pittman etc),
neither in stock or in working solutions.

If you want we can discuss (also offline) possible causes of your failing and perhaps sharing practical aspects of semithin plastic section staining
[which I routinely used until now from 1983 for resins Epon 812(original), Epon-substitute Glycidether 100, LX112**, EMBED 812**, also SPURR's**.. making standardized stock and working solutions since then] could/would be also interesting scientifically (:-).

[NB: * AMbF= Azure II-Methylene blue-basic Fuchsin; ** experiences on externally provided resin blocks/sections.
Disclaimer: No affiliation with or financial interest/support from supplying companies/dealers]

best regards
Wolfgang

Wolfgang MUSS
EM-Lab,
Pathology, SALK-LKH (Gen. Hosp.) & PMU SALZBURG
AUSTRIA

Von: tina-at-pbrc.hawaii.edu [mailto:tina-at-pbrc.hawaii.edu]
Gesendet: Mittwoch, 30. April 2014 03:47
An: Muß Wolfgang
Betreff: [Microscopy] Re: Richardson's Stain for Plastic Sections

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Hi, Vickie-

I use Richardson's all the time.
Try making up fresh components:

1% methylene blue in 1% sodium borate
and
1% Azure II in distilled water

I keep the components in dark brown bottles for several years. When I need
some, I mix them 1:1 (5 ml each) and suck the stain up into a 10ml syringe
fitted with a 0.45 or 0.2 micrometer filter. This way, when you stain
slides it's always freshly filtered. I stain for 40 sec at 60C. It seems
to keep OK in the syringe for weeks to months, and the individual
components in the brown bottles for years.

Aloha,
Tina

} Message: I am trying to stain plastic sections (LR White, Lowicryl and Embed 812 with Richardson's
} stain (1960's)and I have precipitate in the solution.
}
} The precipitates are too large for filtration. Does anyone know what will solubilize them and allow
} for the tissues/cells to take up stain within these plastic ultrathin sections?
}
} I look on the light microscope to see what kinds of cells I have as a preliminary region for grids
} on the TEM.
}
} Thanks!
} Vickie
}
} Login Host: 141.217.26.94
} Listserver Email Form V - 20120416
} ---------------------------------------------------------------------------
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} Do not reply to this message it is from
} the Microscopy Listserver NO-REPLY forwarding
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} Microscopy-at-Microscopy.com
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****************************************************************************
* Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu *
* Biological Electron Microscope Facility * (808) 956-6251 *
* University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf*
****************************************************************************

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From meds-best8-at-skyww.net Wed Apr 30 02:46:45 2014
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Hi Vickie

let's take for granted that your lab has a centrifuge.
I am confident that these large precipitates can be easily pelleted.
I would still filter the supernatant though.

Regards,
Stephane

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Email: vakimler-at-med.wayne.edu
Name: Vickie Kimler

Organization: Wayne State University School of Medicine

Title-Subject: [Filtered] Richardson's Stain for Plastic Sections

Message: I am trying to stain plastic sections (LR White, Lowicryl and Embed 812 with Richardson's
stain (1960's)and I have precipitate in the solution.

The precipitates are too large for filtration. Does anyone know what will solubilize them and allow
for the tissues/cells to take up stain within these plastic ultrathin sections?

I look on the light microscope to see what kinds of cells I have as a preliminary region for grids
on the TEM.

Thanks!
Vickie

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realname - james kammall
Email - parsa319-at-yahoo.com
EDUCATION - Graduate College
SUBJECT_OF_QUESTION - Gilsonite ashes
QUESTION - Dear Sir
I have a project in Gilsonite mine.As you konw Gilsonite,Asphaltum is a
natural, resinous hydrocarbon.
Indeed, the mine owner is going to refine raw Gilsonite and product pure
Gilsonite and
related products. I need to access to knowledge and technology that
enable us to remove Ash
and other additional material from Gilsonite.
Please guide me How to remove Ash from raw gilsonite or refine raw
gilsonite.

Thanks a lot.

James Kammli

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Email: shard-at-uwm.edu
Name: Steve Hardcastle

Organization: UW Milwaukee

Title-Subject: [Filtered] EPI Lamps BH2 Olympus

Message: The lamp/power supply for our epi-lamp for BH2 microscope is kaput.

What are the best alternatives for replacement?
Do the LED lamp sources fit and work ($400)?

steve

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Name: Kerry South

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Title-Subject: 2015 Meeting

Message:

2015 MICROSCOPY NEW ZEALAND CONFERENCE
2-6 February 2015, Dunedin

This biennial conference for microscopists will be held in Dunedin from
Monday the 2nd to Wednesday the 4th of February 2015. It will be
immediately followed by workshops scheduled for the 5th and 6th of
February.

Online registration and abstract submission are now open please see the
event website: Microscopy2015.otago.ac.nz

This event will be hosted by the Otago Centre for Electron Microscopy and
the Otago Centre for Confocal Microscopy at the University of Otago on
behalf of Microscopy New Zealand.

Questions? Interested in becoming an exhibitor or sponsor?
Kerry South, Event Co-ordinator
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From popular-pharmacy13-at-zsttk.ru Wed Apr 30 23:32:02 2014
Return-Path: {popular-pharmacy13-at-zsttk.ru}
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Message-Id: {201405010432.s414W0XU031962-at-ns.microscopy.com}

Has anyone tried the Ultra Bed low viscosity embedding resin? I stumbled
onto this on the EMS site (cat #14310). Looks like it could be a good
replacement for Spurr's - 2 part instead of 4 part, lower viscosity than
the current version of Spurr's.

Phil
--
Philip Oshel
Microscopy Facility Supervisor
Biology Department
024C Brooks Hall
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576

==============================Original Headers==============================
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Listers,

Our met lab produces wonderful EBSD-quality specimens using standard 32 mm epoxy pots. However, I've got some specimens of porous ceramic I want to heat-treat after polishing: breaking out the specimens from epoxy would leave residual polymer in the pores, which I don't want. (Assuming I can break them from the epoxy without shattering the specimens.)

Any thoughts on how to get samples to release from epoxy pots? I'm considering embedding in superglue or wax prior to giving them to the met lab, and then using acetone to float them free from the epoxy pots later. Any thoughts on if this will work? Any better plans? The samples need to be small (10x5x0.5 mm), so I don't want to make our tech hand-polish them if I can avoid it.

Thanks
Chad

---------------------
Chad M. Parish, Ph.D.
Research and Development Staff Member
Materials Science and Technology Division
Oak Ridge National Laboratory
Phone: 1 865 574 0092
Email: parishcm-at-ornl.gov

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8, 36 -- Subject: Metallography: release agent for epoxy pots?
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On Apr 28, 2014, at 4:14 AM, Z.Zhou-at-lboro.ac.uk wrote:

} Dear friends,
} I’m preparing a poster (A1 size) on transmission electron microscopy
} (TEM) topic to decorate our characterisation centre. We have two
} TEMs a Jeol 2000FX and a FEI Tecnai F20 along with typical optical
} microscopes and SEMs, FIB plus surface analysis equipment. Our major
} interests are materials and physical sciences teaching and research.
} My manager suggested some entry level information in the poster for
} public visitors including 16yrs old and their parents. I used to go
} to conferences with research posters. It wasn’t as difficult as this.
} Can you give me some inspirations please? Any suggestions, text or
} websites etc are all appreciated.
} Best regards,
} Zhou
}
Dear Zhou,
You might also want to check out Project MICRO at http://www.microscopy.org/education/projectMICRO
.
Yours,
Bill

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Message: The Central States Microscopy and Microanalysis Society is gathering on Thursday May 22nd,
2014 at MRI Global in Kansas City, Missouri for our spring meeting entitled Â“MacGyver MicroscopyÂ”.
Hopefully everyone remembers the TV show MacGyver, and how ingenious he was repurposing common items
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From cheapest-pharmacy1-at-singnet.com.sg Fri May 2 04:07:47 2014
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Email: margaret.bisher-at-gmail.com
Name: Margaret Bisher

Organization: University of Pittsburgh School of Medicine

Title-Subject: [Filtered] Dendra Fluorophore

Message: Does anyone have an experience using this? I was wondering if it is possible that Dendra
fluorophore might be quenched when trafficked into the acidic compartment of the cytoplasm.

Would you also have any literature reference regarding this?

Thank you, Margaret Bisher

Margaret Bisher
University of Pittsburgh
School of Medicine
Department of Cell Biology
S346 Biomedical Science Tower
Pittsburgh, PA 15213

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Dear All,
anybody out there who can email me a PDF copy of an x-ray examination protocol (dosimetric measurement) for the JEOL 35C SEM?
Best would be, from somewhere in Germany...

Thanks,
Stefan

--

-----------------------------------------------------
Stefan Diller - Scientific Photography
Arndtstrasse 22
D - 97072 Wuerzburg Germany
++49-931-7848700 Phone
++49-931-7848701 Fax
++49-175-7177051 Mobile

Websites:
www.nanoflight.info
www.stefan-diller.com
www.electronmicroscopy.info
www.elektronenmikroskopie.info
www.zwillingsprojekt.de
www.assisi.de
Anfahrt: http://Mail.map24.com/Stefan.Diller
-----------------------------------------------------

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Hi

Some times ago, as we polished ourself single cristal slices for surface
science studies, we embed them in a polyster resin instead of epoxy, to
be able to get them free after polishing. It was a bi-component resin
for metallography (Sody 33, sold by ESCIL in France), which polymerise
at room temperature and is disolved by aceton. I suppose you could get
similar products in the USA by Sturers, Buhler or others.
It's less hard and has more retraction than epoxy, but it works.

After polishing, one put the resin block in aceton during the night, and
one find it as a jely the following morning. It's then easy to pick up
the sample and after 1 or 2 washing in aceton, the samples were clean
enough to be mounted on the sample holder for our study in UHV. They
were not porous, but as far I remember, I think that washing them in
aceton in a semi closed vessel at 40°C in an ultrasonic bath should do
the job for a good cleaning.

An other way would be to try to dissolve your epoxy using the "Strip R"
solvent mix sold by Epotech. But it's not a very nice product, toxic for
the user and complicate to evacuate after use...

Hope it helps

Jacques

J. Faerber
IPCMS-DSI
Institut de Physique et Chimie des Matériaux de Strasbourg
Département Surfaces et Interfaces
23, rue de Loess ; BP43
67034 Strasbourg CEDEX 2
France

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

Le 02/05/2014 01:23, parishcm-at-ornl.gov a écrit :
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} Listers,
}
} Our met lab produces wonderful EBSD-quality specimens using standard 32 mm epoxy pots. However, I've got some specimens of porous ceramic I want to heat-treat after polishing: breaking out the specimens from epoxy would leave residual polymer in the pores, which I don't want. (Assuming I can break them from the epoxy without shattering the specimens.)
}
} Any thoughts on how to get samples to release from epoxy pots? I'm considering embedding in superglue or wax prior to giving them to the met lab, and then using acetone to float them free from the epoxy pots later. Any thoughts on if this will work? Any better plans? The samples need to be small (10x5x0.5 mm), so I don't want to make our tech hand-polish them if I can avoid it.
}
} Thanks
} Chad
}
} ---------------------
} Chad M. Parish, Ph.D.
} Research and Development Staff Member
} Materials Science and Technology Division
} Oak Ridge National Laboratory
} Phone: 1 865 574 0092
} Email: parishcm-at-ornl.gov
}
}
}
}
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} 8, 36 -- Subject: Metallography: release agent for epoxy pots?
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10, 26 -- From jacques.faerber-at-ipcms.u-strasbg.fr Mon May 5 03:27:18 2014
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From drugstore-best9-at-bstar.net Mon May 5 09:01:56 2014
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-----------------------------------------------BIO-Rad Radiance 2100 Laser Scanning Confocal Microscope------------------------

This is a complete and fully functional system. It has always been under service contract, since its purchase in 2005. The reason we are selling the microscope is to make space for other systems.

The confocal system has 7 laser lines (405 nm, 457 nm, 477 nm, 488 nm, 514 nm, 543 nm, and 637 nm). It is attached to a Nikon E800 upright microscope, which has a motorized XY, and a motorized Z stage. The microscope also includes four objectives 10X, 20X, 40X (oil) and 60X (oil).

Asking price: $35,000 (this is essentially the price for a Nikon E800 microscope).

Please contact me offline (307-766-3038, zzhang-at-uwyo.edu), if you are interested, or have any questions.

Thanks,

Zhaojie Zhang
University of Wyoming

==============================Original Headers==============================
8, 33 -- From ZZhang-at-uwyo.edu Mon May 5 14:48:47 2014
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8, 33 -- To: "microscopy-at-microscopy.com" {microscopy-at-microscopy.com}
8, 33 -- Subject: FOR SALE: BIO-Rad Radiance 2100 Laser Scanning Confocal Microscope
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Colleagues

Just a reminder. Using the Listserver to "sell" items
is against our rules. You may post items which are to
be donated, or to be given away for packing and shipping costs.
This forum is not a site to sell used equipment.

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From drugstore_popular7-at-sti-travel.com Tue May 6 02:06:22 2014
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Email: 12hy1-at-queensu.ca
Name: Hongbing Yu

Organization: Queen's University

Title-Subject: [Filtered] TEM Diffraction pattern calibration

Message: Dear all,
I am a PhD student from Queen's University. We have a new Tecnai Osiris TEM in our lab. We tried to
calibrate the diffraction pattern with Standard Mo3O specimen. It was found that the long edge
corresponds to the long g vector(001). According to TEM textbook edited by William, there is no
rotation between diffraction pattern and image. However when we go to HRTEM, we just found that the
FFT for HRTEM rotates 90 degree relative to the diffraction pattern (the sample we use is a HCP
structure metal). Could anyone give me some idea to figure out the 'conflict'?

Best wishes

Hongbing

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Email: 12hy1-at-queensu.ca
Name: Hongbing Yu

Organization: Queen's University

Title-Subject: [Filtered] Accutroller Model 902 required

Message: Dear all,
We need a Accutroller Model 902 for gatan heating holder. If anyone who has a spare used one one for
sale, please let me know and we can pay for it.
Kind regards

Hongbing

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Hello all,

I am in need of a specimen holder for an oldie. A JEOL 100S TEM. It's a two part holder, an it's old enough that I'm hoping someone has one collecting dust on a shelf somewhere.

Thanks in advance!

--Justin A. Kraft

==============================Original Headers==============================
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On May 6, 2014, at 5:29 AM, microscopylistserver-
noreply-at-microscopy.com wrote:

} Name: Hongbing Yu
}
} Organization: Queen's University
}
} Title-Subject: [Filtered] TEM Diffraction pattern calibration
}
} Message: Dear all,
} I am a PhD student from Queen's University. We have a new Tecnai
} Osiris TEM in our lab. We tried to
} calibrate the diffraction pattern with Standard Mo3O specimen. It
} was found that the long edge
} corresponds to the long g vector(001). According to TEM textbook
} edited by William, there is no
} rotation between diffraction pattern and image. However when we go
} to HRTEM, we just found that the
} FFT for HRTEM rotates 90 degree relative to the diffraction pattern
} (the sample we use is a HCP
} structure metal). Could anyone give me some idea to figure out the
} 'conflict'?
}
} Best wishes
}
} Hongbing
}
Dear Hongbing,
Most scopes arrange the lens currents so that the image and
diffraction modes have no rotation for as much of the magnification/
camera length range as practical. For the mags at the very low and
very high ends the manual should list the rotations for both image and
diffraction, from which it is easy to determine the rotation between
the two modes. If that gives 90 deg, there is no 'conflict'.
Yours,
Bill

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Hi colleagues,

This is probably a topic discussed at this list-server before: TEM magnetic materials. The University of Delaware has substantial research in this area and multiple groups need TEM support for magnetic material characterizations. We do take some precautions but still occasionally run into issues. Instead of specifying more details, I would like to know what options or policies your labs have or implement in dealing with magnetic particles, thin films, or foils from bulk magnets?

Many thanks in advance!

Best regards,

*******************************************************
Chaoying Ni, PhD
Associate Professor, Materials Science and Engineering
Director, W. M. Keck Center for Advanced Microscopy and Microanalysis
http://eml.masc.udel.edu; http://www.mseg.udel.edu

Facility location:
Interdisciplinary Science and Engineering Laboratory
221 Academic Street, Newark, DE 19716
http://www.udel.edu/iselab
(302) 831-6359 (Phone); (302) 831-4545 (Fax)

Mailing address:
University of Delaware
201 duPont Hall, Newark, DE 19716
*******************************************************

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9, 27 -- Subject: TEM magnetic materials - multiuser facility operation rules or
9, 27 -- policy options
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X-from: boris.reznik-at-kit.edu ()

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Email: boris.reznik-at-kit.edu
Name: Boris

Organization: KIT

Title-Subject: [Filtered] SAED-Software

Message: Dear all,

we are looking for a suitable software allowing to calculate SAED spot patterns.
The aim of our investigation is to determine the zone axis of minerals.
Using an experimental SAED pattern three diffraction vectors and angles between them are measured.
These parameters should be compared with the calculated one by taking into account the lattice
constants, space group and extinction conditions of the studied crystal.

Do you know a software allowing to solve this task?

With regards
Boris

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Good morning listers,

We recently acquired a Fischinone dual-axis holder (model 2040) for
room-temperature tomography. To our surprise, this holder can only be
tilted (+/- 60 degree) for the very center of grid without touching
pole-pieces. We have used a 2020 holder so far and are extremely happy
with it. We use it on a FEI Tecnai F20 with STEM configuration (X-twin
lenses).

Does anyone encounter this issue before? If yes, how did you get around it?

Thank you in advance for your help.

Xinran Liu

________________________________________

Xinran Nick Liu, M.D. & Ph.D.
Director of Electron Microscopy Core Facility
Research Scientist in Cell Biology
Yale University School of Medicine
333 Cedar Street, SHM I-E16A
New Haven, CT 06520
Office: 203.785.4050
Lab: 203.785.5390
http://medicine.yale.edu/ccmi/em

==============================Original Headers==============================
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Â
I use Single Crystal. It works very well for many cases. It is not really the tool for indexing a completely unknown crystal. It has some zone searching features, but that can still require elbow grease.

Pros: If you have a pretty good idea what the crystal is, or you know it is one of a few (I usually do EDS first so I do have a decent idea what I'm looking at) then this is hard to beat because you can rotate the crystal live. (Also great for getting a feel of the reciprocal space of a given crystal BEFORE your TEM session so you go quickly to a good zone.) You can easily see relative goniometer positions. If you know what a specific reflection is, you can tilt the crystal orthogonal to it to find a specific zone axis, so it has been useful in recognizing that sometimes there are similar looking zones and with other software you may get fooled. For ALCHEMI, you can see when you are aligned along a certain column of atoms easily. You can adjust the unit cell parameters easily for comparison between a couple phases.

Cons: Kinematic only. Searching for complete unknown zones is a bit tricky. I recommend you get on a good zone axis when taking your patterns, don't just try to index a [15,3,7] zone (although, I have managed it in the past).

http://www.crystalmaker.com/singlecrystal/

Zack

(Disclosure: no financial interest but I have done some beta testing.)

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Reply: microscopylistserver-noreply-at-microscopy.com microscopylistserver-noreply-at-microscopy.com(mailto:microscopylistserver-noreply-at-microscopy.com)

Dear Xinran,

The X-twin lens was designed not as a 'STEM' configuration but to
maximise EDX performance (without compromising STEM probe size). This
involved modifications to the pole pieces to allow an EDX detector to be
moved closer to the specimen and hence improve the solid angle, as well
as an extra shield on the lower pole piece to reduce backscatter and
stray X-rays (http://dx.doi.org/10.1017/S1431927605505348).

Unfortunately this does mean that these modifications reduce the tilt
range of most holders when compared to a SuperTWIN lens (more standard
for a 'STEM' configuration). The 2040 holder is quite a bit thicker than
the 2020, and will more than likely lead to pole-touch in a narrower
pole piece gap. For your information the 2040 has no tilt restrictions
in a SuperTWIN lens, but maximum useful tilt angle is ~+/- 70 degrees
due to holder shadowing.

As this a geometrical problem I don't see an easy solution. Use the 2020
and take the specimen out and rotate it 90 degrees between dual-axis
tilting would be my suggestion! The loss of information between 60-70
degrees will severely affect reconstruction (much more so than 70-80).
We mostly use our 2040 to orient specimen features perpendicular to the
tilt axis prior to tomography (or as a really high tilt DT holder for
crystallography).

Matthew

On 8/05/2014 12:36 AM, xinran.liu-at-yale.edu wrote:
} Good morning listers,
}
} We recently acquired a Fischinone dual-axis holder (model 2040) for
} room-temperature tomography. To our surprise, this holder can only be
} tilted (+/- 60 degree) for the very center of grid without touching
} pole-pieces. We have used a 2020 holder so far and are extremely happy
} with it. We use it on a FEI Tecnai F20 with STEM configuration (X-twin
} lenses).
}
} Does anyone encounter this issue before? If yes, how did you get around it?
}
} Thank you in advance for your help.
}
} Xinran Liu
}
} ________________________________________
}
} Xinran Nick Liu, M.D. & Ph.D.
} Director of Electron Microscopy Core Facility
} Research Scientist in Cell Biology
} Yale University School of Medicine
} 333 Cedar Street, SHM I-E16A
} New Haven, CT 06520
} Office: 203.785.4050
} Lab: 203.785.5390
} http://medicine.yale.edu/ccmi/em
}
--
Dr M.Weyland, Associate Professor & Titan Manager
--------------------------------------------------------------------------
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Email: bsmit112-at-jhmi.edu
Name: Barbara Smith

Organization: Johns Hopkins University

Title-Subject: [Filtered] Sorvall JB4 Microtome

Message: We recently acquired a Sorvall JB4 microtome that is supposed to do both paraffin and
plastic sectioning. Unfortunately, it came without the paraffin chuck and the blade holder. Does
anyone know of a group or have such an accessory that they would be willing to part with? Or have
any suggestions on how to modify? Thank you.

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I am required to eliminate all Windows XP systems and replace them with
Windows 7 systems. I have been unable to determine how this may affect a
JEOL1400 or if new software and drivers are required. Has anyone
completed this OS upgrade on a JEOL1400?
Thanks,
Larry

--
Larry Ackerman, Specialist
Electron Microscopy Lab
Manager, Diabetes & Endocrinology Research Center Microscopy Core
UCSF, Dept. of Anatomy, Rm S657
513 Parnassus Ave., Box 0452
San Francisco, CA 94143

larry.ackerman-at-ucsf.edu

415-999-4758

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We got around this by getting the IT guys to set up a separate VLAN for
our old XP systems. We have at least 3 systems that cannot be upgraded.
This seems to work fine - data can be uploaded to the stand-alone server
and people can then download it to their computers. This could be an
interim solution even if you do eventually convert all to 7.

cheers,
Rosemary

Rosemary White
CSIRO Plant Industry
GPO Box 1600
Canberra, ACT 2601
Australia

T 61 2 6246 5475
F 61 2 6246 5334M 61 2 468 966 713
E rosemary.white-at-csiro.au

On 8/05/14 11:16 AM, "larry.ackerman-at-ucsf.edu" {larry.ackerman-at-ucsf.edu}
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From drugstore_reasonable14-at-js-llp.com Thu May 8 00:39:30 2014
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Name: Roger Ristau

Organization: Univ of Connecticut

Title-Subject: [Filtered] cleaning evaporator

Message: Does anyone have an effective method for cleaning metal deposition from the inside of an
evaporator bell-jar?

Cheers
Roger Ristau
University of Connecticut

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Email: CGoldsmith-at-cdc.gov
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Organization: Centers for Disease Control and Prevention

Title-Subject: [Filtered] Osmium Tetroxide SOP

Message: Our Safety Office would like for us to write up an SOP in case of a spill of osmium
tetroxide. I thought that there is no need to reinvent the wheel, and would like to ask for other
laboratories who already have an SOP written up if they would share it with me, offline at
CGoldsmith-at-cdc.gov.

Thank you,

Cynthia Goldsmith, MGS
Infectious Diseases Pathology Branch
Centers for Disease Control and Prevention
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I use Bon Ami, a non-abrasive kitchen cleaner, a damp paper towel, and
scrub. Nothing else.

Phil

} Email: roger.ristau-at-uconn.edu
} Name: Roger Ristau
}
} Organization: Univ of Connecticut
}
} Title-Subject: [Filtered] cleaning evaporator
}
} Message: Does anyone have an effective method for cleaning metal deposition from the inside of an
} evaporator bell-jar?
}
} Cheers
} Roger Ristau
} University of Connecticut
--
Philip Oshel
Microscopy Facility Supervisor
Biology Department
024C Brooks Hall
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576

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All,

We have a Phillips (fei) XL40 ESEM (circa 2000) with the latest
microscope control software. The installed EDAX Phoenix EDS system is
failing. The x-ray detector (EDAX detecting unit PV7761/53ME) seems to
be functioning properly. An EDAX Genesis system has been located and we
would appreciate any information members of the group have regarding:

1) Compatibility of the installed x-ray detector with EDAX Genesis.

2) Configuring the serial interface between EDAX Genesis and fei's
microscope control.

3) Any gotchas...

Thank you,

Owen Mills
Michigan Tech

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Also, Phil's message suggesting using Bon Ami, damp paper towels and scrubbing is in line with my experience.

I know it is obvious, but do not leave water or cleaner residue in the chamber.

Dan

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Email: roger.ristau-at-uconn.edu
Name: Roger Ristau

Organization: Univ of Connecticut

Title-Subject: [Filtered] cleaning evaporator

Message: Does anyone have an effective method for cleaning metal deposition from the inside of an evaporator bell-jar?

Cheers
Roger Ristau
University of Connecticut

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We have a Leitz LEO 1450 SEM that we are trying to get up and running. Does
anyone know where we can get a manual? Or, pay to have one copied?

Gordon Whitlock
Glow Research
Phoenix, AZ USA
cell 480-710-6469
gordonwhitlock-at-glowresearch.org

Gordon Whitlock
VP Sales
Trion Technology
1025 S. 52nd St.
Tempe, AZ. 85281
USA 480-968-8818 X-15
cell 727-214-7189

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This may not help you with current cleaning but will with subsequent ones....

After the bell jar has been cleaned, we have had good luck using a thin layer of dish soap on the inside of the bell jar which allows the next cleaning to relatively easy as the metal coating will release much easier with warm water. The procedure we use is to clean the bell jar with warm water to remove the prior coating, then using a small drop of dish soap and rubber gloves, smooth it around the inside of the bell jar until it is a white film and allow to dry for a little while. Then replace the bell jar and pump down to outgas... I usually run a "dummy" coating to make sure all is well...

Regards,
Scott

Scott A. Payne
Assistant Director
Electron Microscopy Center
North Dakota State University
Phone: 701.231.8234

Mailing Address Physical Address/Courier
USDA-ARS NCSL, USDA-ARS NCSL
1605 Albrecht Blvd 1307 North 18th St.
Fargo ND 58102-2765 Fargo, ND 58102

http://www.ndsu.edu/em_lab/

-----Original Message-----
X-from: oshel1pe-at-cmich.edu [mailto:oshel1pe-at-cmich.edu]
Sent: Thursday, May 08, 2014 9:31 AM
To: Payne, Scott

I use Bon Ami, a non-abrasive kitchen cleaner, a damp paper towel, and scrub. Nothing else.

Phil

} Email: roger.ristau-at-uconn.edu
} Name: Roger Ristau
}
} Organization: Univ of Connecticut
}
} Title-Subject: [Filtered] cleaning evaporator
}
} Message: Does anyone have an effective method for cleaning metal
} deposition from the inside of an evaporator bell-jar?
}
} Cheers
} Roger Ristau
} University of Connecticut
--
Philip Oshel
Microscopy Facility Supervisor
Biology Department
024C Brooks Hall
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576

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18, 34 -- From Scott.Payne-at-ndsu.edu Thu May 8 10:24:07 2014
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Fullam used to sell a product called "Victawet" that you evaporated onto the surface to make cleaning easier.

Dan

-----Original Message-----
X-from: Scott.Payne-at-ndsu.edu [mailto:Scott.Payne-at-ndsu.edu]
Sent: Thursday, May 08, 2014 9:37 AM
To: Crane, Dan - OSHA

This may not help you with current cleaning but will with subsequent ones....

After the bell jar has been cleaned, we have had good luck using a thin layer of dish soap on the inside of the bell jar which allows the next cleaning to relatively easy as the metal coating will release much easier with warm water. The procedure we use is to clean the bell jar with warm water to remove the prior coating, then using a small drop of dish soap and rubber gloves, smooth it around the inside of the bell jar until it is a white film and allow to dry for a little while. Then replace the bell jar and pump down to outgas... I usually run a "dummy" coating to make sure all is well...

Regards,
Scott

Scott A. Payne
Assistant Director
Electron Microscopy Center
North Dakota State University
Phone: 701.231.8234

Mailing Address Physical Address/Courier
USDA-ARS NCSL, USDA-ARS NCSL
1605 Albrecht Blvd 1307 North 18th St.
Fargo ND 58102-2765 Fargo, ND 58102

http://www.ndsu.edu/em_lab/

-----Original Message-----
X-from: oshel1pe-at-cmich.edu [mailto:oshel1pe-at-cmich.edu]
Sent: Thursday, May 08, 2014 9:31 AM
To: Payne, Scott

I use Bon Ami, a non-abrasive kitchen cleaner, a damp paper towel, and scrub. Nothing else.

Phil

} Email: roger.ristau-at-uconn.edu
} Name: Roger Ristau
}
} Organization: Univ of Connecticut
}
} Title-Subject: [Filtered] cleaning evaporator
}
} Message: Does anyone have an effective method for cleaning metal
} deposition from the inside of an evaporator bell-jar?
}
} Cheers
} Roger Ristau
} University of Connecticut
--
Philip Oshel
Microscopy Facility Supervisor
Biology Department
024C Brooks Hall
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576

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I agree that a thin layer of detergent is good, but my experience is
that dish-washing detergent is better than dish soap.

And this gives me the opportunity to get off track for a moment. The
main purpose of shaving foam is to make the skin wet. So I shave after
showering with no shaving soap - it works just fine. However the mirror
tends to get steamed up during the shower. The solution to this problem
is to apply a little dish-washing fluid to the mirror. Then it does not
steam up.
--
Alwyn Eades
Department of Materials Science and Engineering
Lehigh University

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Dear Boris,

to this target I use CaRIne Crystallography Software

http://carine.crystallography.pagespro-orange.fr/

Ciao,

Elena

Elena BELLUSO

Dip. di Scienze della Terra, Universita' degli Studi di Torino

Via Valperga Caluso n. 35, 10125 TORINO - Italy

tel: (39) 011 670 51 35 - fax: (39) 011 670 51 28

e-mail: elena.belluso-at-unito.it

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I spray the inside of the bell jar or cylinder with unscented White Rain
hair spray. Warm water will dissolve the hairspray and release the metal,
whether evaporated or sputtered. Using unscented avoids unneeded volatiles
in the vacuum system.

Ken Converse
owner

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

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X-from: jae5-at-lehigh.edu [mailto:jae5-at-lehigh.edu]
Sent: Thursday, May 08, 2014 11:59 AM
To: kenconverse-at-qualityimages.biz

I agree that a thin layer of detergent is good, but my experience is
that dish-washing detergent is better than dish soap.

And this gives me the opportunity to get off track for a moment. The
main purpose of shaving foam is to make the skin wet. So I shave after
showering with no shaving soap - it works just fine. However the mirror
tends to get steamed up during the shower. The solution to this problem
is to apply a little dish-washing fluid to the mirror. Then it does not
steam up.
--
Alwyn Eades
Department of Materials Science and Engineering
Lehigh University

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From pharmacy_canadian7-at-airtelbroadband.in Thu May 8 13:38:58 2014
Return-Path: {pharmacy_canadian7-at-airtelbroadband.in}
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We use Jif (known as Cif in some places). It is a non-abrasive cleaner probably exactly like Bon Ami.

After years (decades in my lab's history) of using Wenol to clean parts of our TEM gun etc., we were first surprised then tried it on the advice of a senior FEI engineer.

The main ingredients are calcium carbonate particles and a couple of surfactants. As long as the scented versions are avoided, it leaves no residue if rinsed properly since all ingredients are very water soluble.

Duane

Duane Harland, Agresearch, New Zealand

________________________________________
X-from: oshel1pe-at-cmich.edu [oshel1pe-at-cmich.edu]
Sent: 09 May 2014 02:39
To: Harland, Duane

I use Bon Ami, a non-abrasive kitchen cleaner, a damp paper towel, and
scrub. Nothing else.

Phil

} Email: roger.ristau-at-uconn.edu
} Name: Roger Ristau
}
} Organization: Univ of Connecticut
}
} Title-Subject: [Filtered] cleaning evaporator
}
} Message: Does anyone have an effective method for cleaning metal deposition from the inside of an
} evaporator bell-jar?
}
} Cheers
} Roger Ristau
} University of Connecticut
--
Philip Oshel
Microscopy Facility Supervisor
Biology Department
024C Brooks Hall
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576

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Email: willis.robert-at-epa.gov
Name: Bob Willis

Organization: US EPA

Title-Subject: [Filtered] Bruker EDS-Tescan interface problem

Message: I have a Quantax EDS system attached to a Tescan MIRA3 FE-SEM. The two PCs are
communicating with each other, but the Quantax is not able to take control of the SEM or read SEM
parameters (mag, HV, etc). It is capable of collecting an image if the SEM is put in external
scanning mode.

Thanks for any suggestions.

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Email: roger.ristau-at- uconn.edu
Name: Roger Ristau

Organization: Univ of Connecticut

Title-Subject: [Filtered] cleaning evaporator

Message: Who would have thought that such an innocent question would bring so many interesting
solutions. Thanks to all for great ideas. If the improvement in my results is as good as suggested,
I may quit my day job and go into the "window washing" business!

Cheers
Roger Ristau
University of Connecticut

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It has been a long time since I cleaned a bell jar, but I seem to
recall using Bon Ami, as others suggest. However, to facilitate
removal of subsequently deposited materials I seem to remember
evaporating a fair size piece of rock salt onto the inside surface
right after cleaning. This is a non-organic material with low vapor
pressure, and, as I recall, did a pretty good job as a release agent
for the next cleaning,
--
Wilbur C. Bigelow, Professor Emeritus
Materials Sci. & Engr., Univ. of Michigan
Ann Arbor, Michigan 48109-2136
e-mail: bigelow-at-umich.edu;
Fx:734-763-4788; Ph:734-975-0858
Address mail to: 2911 Whittier Court
Ann Arbor, MI 48104-6731

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Hi All
If you are interested in Outreach and putting on a microscopy workshop at
a local school or community then you will be interested in the programs on
offer this year at the M&M meeting in Hartford in August. Microscopy in
the Classroom has a number of talks by experienced speakers and
ProjectMICRO is putting on a hands-on workshop for delegates and teachers.
All participants will take away experiences and ideas to enhance their
program. I have put flyers for these two sessions in a dropbox that you
can download at
https://www.dropbox.com/sh/fkyls0q5jny69ah/AACrHMOijcXm6fF9EXbLYtP7a

Also we are planning another Family Affair for delegates families and
friends. Each year we try to bring your favourites and something
different. This year....

The story so far
A secret agent, James B Atom, wrote a message very very very small. The
first part of the message can be read with a light microscope. The second
part of the message can only be read using a scanning electron microscope.
The message gets split up and passed on and part comes to you. Your
mission, should you take it, is to decipher the message, and bring it to
headquarters (the Outreach booth). To assist you in this, we have the use
of the devices needed in the classroom and the Exhibition Hall.

Hope to see you all in August
Elaine

Dr. Elaine C. Humphrey
STEHM Technologist and Lab Manager
Bob Wright Science Centre A015
Advanced Microscopy Facility
University of Victoria, Canada

Lab: 250-853-3968
website: http://www.stehm.uvic.ca

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For those of you who do color brightfield, this might be an
interesting seminar.

"Achieving Color Integrity in Brightfield Images: A Color Science
Method for Color Calibration"
When: Tuesday, May 13, 2014 -at- 10:00am
Where: Health Science Research Facility, University of Vermont
College of Medicine, Burlington, VT

Hosted by the Microscopy Imaging Center, Datacolor will present a
seminar at the University of Vermont College of Medicine that reviews
opportunities to maintain color integrity in brightfield imaging,
beginning with sources of color variability and offering a new
approach to making color brightfield images publication ready without
using Photoshop.

Abstract: Transmitted light brightfield microscope images reveal the
impact of an enormous number of color variables outside the specimen
itself. This seminar will review the sources of many of these
variables, and discuss mitigating approaches to controlling color
variability in brightfield images. A novel color science-based
solution offers an empirical approach to calibrating images to a
standard for greater consistency, comparability and efficiency than
existing methods, from image capture through display. Additionally,
it automatically records the audit trail, a considerable advantage
when presenting images to reviewers.

For further details, visit www.ChromaCal.com

Best regards,
Barbara Foster, President & Chief Consultant
Microscopy/Microscopy Education*
www.MicroscopyEducation.com

*A subsidiary of The Microscopy & Imaging Place, Inc.
7101 Royal Glen Trail, Suite A
McKinney, TX 75070
P: 972-924-5310
F: 214-592-0277

MME is currently scheduling courses for now and through Fall 2014.
Call us today for a free training evaluation.

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Email: gary-at-gaugler.com
Name: Dr Gary Gaugler

Organization: Microtechnics, Inc.

Title-Subject: [Filtered] FEI Quanta 600 ESEM

Message: Greetings. I'm considering purchasing a Quanta ESEM (gen 1, W) and wonder if there are
downsides to this for general SEM work? Usually I have done high vac imaging using SFEG SEMs. The
Quanta looks to be really different from high vac only tools and also from variable pressure tools.
Can it really do a good job of different specimens and in different modes up to about 100KX? Would
appreciate hearing from any users about hints and experiences.

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Organization: Microscopy Society of Northeastern Ohio, MSNO

Title-Subject: [Filtered] Spring MSNO meeting - May 21st Meeting at John Carroll University

Message: Spring MSNO meeting - May 21st Meeting at John Carroll University

We have updated our website with the full program information.
http://www.msneo.org/meetings.html
If you plan to join please register ASAP.

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We replaced a venerable JEOL 840A (W-gun) with a FEI Quanta FEG 250 about 4 years ago. We have not been disappointed.

For several years we had been imaging nanoparticles at 50kx on the 840 and figured we really needed to make the switch to a field emission gun to get better resolution. Most application engineers told us we were doing quite well considering our W gun, but there was no doubt about the performance improvement to come from the FE gun (1 nm vws. 3 nm resolution).

I suppose the Quanta with tungsten gun should be good, but it may be a stretch to get satisfying images at 100kx. If you want, I can send you some comparison images.

Of course, even though we can reach 100kx rather easily on the Quanta-FEG, our users also spend a lot of time below 1kx. (I have to wonder why they are using this scope instead of a W alternative.)

Warren
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Email: gary-at-gaugler.com
Name: Dr Gary Gaugler

Organization: Microtechnics, Inc.

Title-Subject: [Filtered] FEI Quanta 600 ESEM

Message: Greetings. I'm considering purchasing a Quanta ESEM (gen 1, W) and wonder if there are
downsides to this for general SEM work? Usually I have done high vac imaging using SFEG SEMs. The
Quanta looks to be really different from high vac only tools and also from variable pressure tools.
Can it really do a good job of different specimens and in different modes up to about 100KX? Would
appreciate hearing from any users about hints and experiences.

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Email: patricia.bozzano-at-gmail.com
Name: PATRICIA

Title-Subject: [Filtered] ISO 17025 Acreditation

Message: Dear Listers,
We are working to get ISO 17025 accreditation for our scanning electron microscopy laboratory.
I do appreciate so much if someone who reached the accreditation can help us with the sample
selection to calculate the accuracy of the method.
Thanks you in advance,
Yours,
Patricia

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Email: joanne.etheridge-at-monash.edu
Name: Joanne Etheridge

Organization: Monash University

Title-Subject: [Filtered] Professor of Structural Cryo-Electron Microscopy - position available

Message: Position Available:

The Monash-Warwick Professor of Structural Cryo-Electron Microscopy

A joint position with
Monash University, Australia (80%)and Warwick University, UK (20%)

http://jobs.monash.edu.au/jobDetails.asp?sJobIDs=523391

Deadline has some flexibility.

For enquiries, please contact:

Professor James Whisstock
ARC Federation Fellow
Department of Biochemistry
Monash University
Clayton, Melbourne, VIC, 3800
Australia
Email: james.whisstock-at-monash.edu

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I would be interested in this information too so if you wouldn't mind replying to me as well or to the board it would be greatly appreciated. Thanks.

Sincerely,

James Neal-Kababick
Director
Flora Research Laboratories
An FDA and DEA Registered & Inspected Laboratory
Fellow AOAC International
Adjunct Faculty Bastyr University
Botanical Medicine Department
1-541-472-0980 phone
jimk-at-floraresearch.com
www.floraresearch.com

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Sent: Sunday, May 11, 2014 7:42 PM
To: James Neal-Kababick

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Email: patricia.bozzano-at-gmail.com
Name: PATRICIA

Title-Subject: [Filtered] ISO 17025 Acreditation

Message: Dear Listers,
We are working to get ISO 17025 accreditation for our scanning electron microscopy laboratory.
I do appreciate so much if someone who reached the accreditation can help us with the sample selection to calculate the accuracy of the method.
Thanks you in advance,
Yours,
Patricia

==============================Original Headers==============================
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Email: pwebster-at-usc.edu
Name: Paul Webster

Organization: University of Southern California

Title-Subject: [Filtered] Third Party Service for EM

Message: Dear All,

I know this subject comes up often, and I have the same feelings about the subject that are often
expressed on this server. However I can't find the material in the archives. My apologies in advance
for this question.

So, here goes: Does anyone have any experiences with service being provided through third party
companies such as the Remi Group.

Past replies have highlighted the lack of specific support for electron microscopes, the difficulty
in getting priority service, the need to pay for parts etc.

Do these concerns still apply or have the third party providers improved their service? Comments
from anyone who is or has been covered by such a company will be welcome, however,replies from
anyone with an opinion are encouraged (positive and negative). We are preparing our reply to our
bean counters.

Best regards,

Paul Webster, Ph.D.
University of Southern California

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Email: coreimaging-at-umaryland.edu
Name: Ru-ching Hsia

Organization: University of Maryland, Baltimore

Title-Subject: [Filtered] workshop announcement

Message: Dear Colleagues,

We would like to draw your attention that the University of Maryland Baltimore will be hosting the
First Annual Current EM Techniques Workshop on June 2 and 3, 2014. The purpose of this annual
workshop is to introduce participants to new electron microscopy techniques and instruments via live
demonstration and open discussion. The focus of this yearÂ’s workshop is cryo-EM sample preparation
and cryo-EM. The workshop will include oral presentations and tips-and-tricks discussion forums
during the morning session followed by live instrument demonstration in the afternoon session. Demo
instruments will feature high pressure freezer, freeze substitution systems, a cryo-ultramicrotome,
a grid plunger, cryo-TEM and cryo-SEM. A dinner event will be co-sponsored by the Chesapeake
Microscopy and Microanalysis society (CMMS) on June 2nd providing opportunities for social and
scientific exchange among workshop participants and CMMS members.

More information regarding the workshop and registration can be found on our website:
http://www.dental.umaryland.edu/Core-imaging/2010web/events/CurrentEMWorkshop.html .

The registration will be open until May 30th.

For other inquiries send email to: UMBCurrentEMWorkshop-at-umaryland.edu

We look forward to see you in Baltimore.

Core Imaging Facility
University of Maryland, Baltimore

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From discounted-medications5-at-vripack.com Mon May 12 21:23:37 2014
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Dear Paul,

There are not just worlds, but Universes of difference between service
from the third-party providers (some of whom are very good and some are
not so much), and "cost reduction offers" from the middlemen trying to
make a buck by inserting themselves between the end-users and OEM (or
third-party) service providers.

Be very careful and read the finest print on whatever service offering
you are considering...

Valery Ray
=================================
PBS&T, MEO Engineering Co., Inc.
290 Broadway, Suite 298
Methuen, MA 01844 USA
Phone: +1-978-296-5063 - leave a message with call-back number
US Mobile: +1-978-305-0479
Skype: pbstmeo
E-mail: vray-at-partbeamsystech.com
Web: www.partbeamsystech.com
Web: www.freudlabs.com

On 5/12/2014 7:54 PM, microscopylistserver-noreply-at-microscopy.com wrote:
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} Email: pwebster-at-usc.edu
} Name: Paul Webster
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} Organization: University of Southern California
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} Title-Subject: [Filtered] Third Party Service for EM
}
} Message: Dear All,
}
} I know this subject comes up often, and I have the same feelings about the subject that are often
} expressed on this server. However I can't find the material in the archives. My apologies in advance
} for this question.
}
} So, here goes: Does anyone have any experiences with service being provided through third party
} companies such as the Remi Group.
}
} Past replies have highlighted the lack of specific support for electron microscopes, the difficulty
} in getting priority service, the need to pay for parts etc.
}
} Do these concerns still apply or have the third party providers improved their service? Comments
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From canadian-meds11-at-airpurifyingsystems.com Tue May 13 04:23:45 2014
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First, I want to thank the members of this list for all the help keeping our donated SEM running for the past year. The advice and connections I've made have been extremely valuable.

Now I'm asking a different kind of question. Some background:

Pumping Station: One is a non-profit "hackerspace" or "makerspace" located in Chicago. We are an organization that provides members with access to all sorts of tools (woodworking, machine shop, electronics, welding, CNC, 3D printing, laser cutting, home brewing and cooking, sewing and machine knitting, etc.) but more than that, we have built a community of around 270 makers working on projects, offering classes, and offering plenty of informal mentoring and support for each other's projects.

So, last year, we received a donation of a Leica S430. We believe we are the only hackerspace with a working SEM. I've been building up infrastructure for a microscopy lab around it. The scope works reliably (even if the vacuum is not as good as I'd like) with a repairable BSD and a working EDX, and we have a working sputter coater, critical point dryer, and ultrasonic cleaner. I'm finishing up a project to have bulk liquid nitrogen available, and it looks like I might have a source for a binocular inspection microscope for sample prep.

Now I'm focusing on the "people" side of things, and what to do with all of this. We're in an unusual situation. Universities and labs have reasons to own a SEM. High schools with donated SEMs have curricula / after school programs. While our organization is educational, it isn't formal in the sense of a school, and a SEM is really an extra or bonus.

I've put together a training program which is popular. People like to learn new tools. I've got more than 20 people authorized to use it on their own. Yet there has been very little use outside of the training; often people don't touch it afterwards. (In fairness, this is true of other tools too, e.g. welding, machining....)

Part of it may be that I have not yet started offering sample prep classes / authorizations, although I've made it clear that I'm happy to prepare samples for people and load them in the chamber. So far only two people have taken me up on that. Admittedly, sample prep is an area I know little about, especially biological sample prep. (Did I mention that I'm pretty much self taught in all this?) The CPD was an impulse buy on eBay, before I realized we need to do glutaraldehyde fixing first which needs a fume hood to do safely, so I'm not sure how to handle wet biological samples. I'm left with a lot of questions. Like whether PLA or ABS plastics outgas at SEM vacuum levels, and whether they could be sputter coated and wouldn't melt under the beam. (We do a lot of 3D printing, and it would be interesting to compare these.) Or how to mount specimens with small particulates. (I made a trip to the Chernobyl exclusion zone, and I'd like to do EDX analysis on samples collected.)

So I'm left puzzled. I think it is unprecedented to have an SEM available 24/7, free of charge with free training, to the general public in the middle of a major city, yet nobody seems to be taking advantage of it. (The general public would have to become members to use it, but membership is open to anyone 18 and older for $40 a month, and that includes access to all the other tools.)

I'm not sure if people are intimidated by it, or if they have a hard time thinking of applications for it. I've considered having open "SEM Salons" where we would have a sample or project to work on as a group. But by and large, I'm looking for ideas of stuff to do, methods for sample prep, ways to do outreach to get people involved, etc.

I'm also looking for groups to partner with. High schools are problematic at this time because we are still trying to nail down liability issues around minors. Collaboration with for-profit groups would be fine too, provided they did participate and give back to our community.

And, of course, if anyone on this list is local to Chicago, we are looking for members to get involved with the work we are doing.

Hackerspaces are a relatively new concept, and while we may be the only one now with a working SEM, I think that might change. I know a hackerspace in the Washington DC area is repairing a Cambridge Stereoscan 200. So I'm very much interested in best practices surrounding open science and public SEM access.

Regards,
Ryan Pierce

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

Dear Friends,
Can you please advise me where I can find a concise (and scientific accurate) definition for transmission electron microscopy? Also for scanning electron microscopy?
Zhaoxia Zhou
Dr Z Zhou
Loughborough University
Leicestershire UK

==============================Original Headers==============================
2, 31 -- From Z.Zhou-at-lboro.ac.uk Tue May 13 10:40:23 2014
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2, 31 -- From: Zhaoxia Zhou {Z.Zhou-at-lboro.ac.uk}
2, 31 -- To: "Microscopy-at-microscopy.com" {Microscopy-at-microscopy.com}
2, 31 -- Subject: definition for TEM and SEM
2, 31 -- Thread-Topic: definition for TEM and SEM
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From meds-popular8-at-rowdyism.com Tue May 13 13:13:22 2014
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Message-Id: {201405131813.s4DIDIwL028380-at-ns.microscopy.com}

We have an old Polaron evaporator - I don't know how old as the labeling is practically non-existent. Anyway, we seem to have lost the solenoid that operates the baffle valve underneath the bell jar. Apparently it has been known to stick and it reportedly took some coaxing to make sure it opened. Anyway, it appears to have been stuck yesterday and it is now rather crispy. Perhaps it is still functional, but it certainly got very hot.

Just in case it is fried, does anyone happen to have one of these laying around that they are not using that we might be able to get parts?

Warren Straszheim
Ames Laboratory
Ames, IA

==============================Original Headers==============================
3, 35 -- From wesaia-at-iastate.edu Tue May 13 13:56:00 2014
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3, 35 -- From: "Straszheim, Warren E [BIOTC]" {wesaia-at-iastate.edu}
3, 35 -- To: MSA {Microscopy-at-microscopy.com}
3, 35 -- Subject: Polaron evaporator woes
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Hi Ryan,

I suggest that with a working SEM and EDS you contact some mineralogical
clubs in the area. At least I would be very interested in running
unknown samples, too bad I live in Sweden.

But, on the other hand I have a TEM and a SEM in my private hackerspace. :-D

I'm partly attached to the local hackerspace here in Umeå, I know
several of the members, but so far I have more toys and bigger space
where I store my "junk" so even if I wanted to I can't donate anything
to them, they don't have the space for it.
I started my journey into the EM world since I collect minerals and
there are always strange things you can't identify with simple means. So
I got a donated TEM, restored it (it took a year) and when I got it
running I promptly melted some asbestos fibres in the electron beam...
there's quite a punch if you focus it good enough!
My plan was to set up a lab to run my own samples and others, not quite
there yet. When I've talked about it with fellow collectors I got quite
a lot of interest. Therefore I suggest you contact mineral societies in
the area, I'm sure there is large interest among them.

I'm in a middle of a move from one storage to a bigger one, and when
that is finished I will bring the microscopes up again. The only thing
I'm lacking is a working EDS. I have a system but the computer is from
1974 and uses core memory, that in it self will be quite a project to
get running. :-D
What I have is a JEOL JEM 100CX TEM with ASID, a JSM-25 SEM, a JEOL
carbon evaporator, and a water chiller. It could be a fun project on
rainy summer days to put everything in working order again after the move.

Regards, Göran

rdpierce-at-pobox.com skrev 2014-05-13 14:51:
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} First, I want to thank the members of this list for all the help keeping our donated SEM running for the past year. The advice and connections I've made have been extremely valuable.
}
} Now I'm asking a different kind of question. Some background:
}
} Pumping Station: One is a non-profit "hackerspace" or "makerspace" located in Chicago. We are an organization that provides members with access to all sorts of tools (woodworking, machine shop, electronics, welding, CNC, 3D printing, laser cutting, home brewing and cooking, sewing and machine knitting, etc.) but more than that, we have built a community of around 270 makers working on projects, offering classes, and offering plenty of informal mentoring and support for each other's projects.
}
} So, last year, we received a donation of a Leica S430. We believe we are the only hackerspace with a working SEM. I've been building up infrastructure for a microscopy lab around it. The scope works reliably (even if the vacuum is not as good as I'd like) with a repairable BSD and a working EDX, and we have a working sputter coater, critical point dryer, and ultrasonic cleaner. I'm finishing up a project to have bulk liquid nitrogen available, and it looks like I might have a source for a binocular inspection microscope for sample prep.
}
} Now I'm focusing on the "people" side of things, and what to do with all of this. We're in an unusual situation. Universities and labs have reasons to own a SEM. High schools with donated SEMs have curricula / after school programs. While our organization is educational, it isn't formal in the sense of a school, and a SEM is really an extra or bonus.
}
} I've put together a training program which is popular. People like to learn new tools. I've got more than 20 people authorized to use it on their own. Yet there has been very little use outside of the training; often people don't touch it afterwards. (In fairness, this is true of other tools too, e.g. welding, machining....)
}
} Part of it may be that I have not yet started offering sample prep classes / authorizations, although I've made it clear that I'm happy to prepare samples for people and load them in the chamber. So far only two people have taken me up on that. Admittedly, sample prep is an area I know little about, especially biological sample prep. (Did I mention that I'm pretty much self taught in all this?) The CPD was an impulse buy on eBay, before I realized we need to do glutaraldehyde fixing first which needs a fume hood to do safely, so I'm not sure how to handle wet biological samples. I'm left with a lot of questions. Like whether PLA or ABS plastics outgas at SEM vacuum levels, and whether they could be sputter coated and wouldn't melt under the beam. (We do a lot of 3D printing, and it would be interesting to compare these.) Or how to mount specimens with small particulates. (I made a trip to the Chernobyl exclusion zone, and I'd like to do EDX analysis on samples collected.)
}
} So I'm left puzzled. I think it is unprecedented to have an SEM available 24/7, free of charge with free training, to the general public in the middle of a major city, yet nobody seems to be taking advantage of it. (The general public would have to become members to use it, but membership is open to anyone 18 and older for $40 a month, and that includes access to all the other tools.)
}
} I'm not sure if people are intimidated by it, or if they have a hard time thinking of applications for it. I've considered having open "SEM Salons" where we would have a sample or project to work on as a group. But by and large, I'm looking for ideas of stuff to do, methods for sample prep, ways to do outreach to get people involved, etc.
}
} I'm also looking for groups to partner with. High schools are problematic at this time because we are still trying to nail down liability issues around minors. Collaboration with for-profit groups would be fine too, provided they did participate and give back to our community.
}
} And, of course, if anyone on this list is local to Chicago, we are looking for members to get involved with the work we are doing.
}
} Hackerspaces are a relatively new concept, and while we may be the only one now with a working SEM, I think that might change. I know a hackerspace in the Washington DC area is repairing a Cambridge Stereoscan 200. So I'm very much interested in best practices surrounding open science and public SEM access.
}
} Regards,
} Ryan Pierce

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Title-Subject: [Filtered] SEM Technician Position

Message: SEM Technician
Our TEM/FIB lab is located in Portland, Oregon and is equipped with a state-of-the-art FEI Osiris
TEM and a Helios 600 FIB/SEM. SEM technician position will focus on cross-sectional SEM
characterization and TEM sample preparation. This SEM opening needs to be filled in immediately.
Term:
Must be available for a minimum of 3 months. Term could be extended to 6-12 months.
Work Schedule:
Requires flexibility to work on night or weekend shifts to accommodate customersÂ’ urgent SEM requests.
Education:
Associate degree or equivalent combination of education and experience in Engineering, Materials
Science, Chemistry, or Physics, etc.
Experience:
Hands-on expert in SEM, FIB, and dual beams. Hands-on experience on TEM sample preparation a plus.
Skills:
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Name: Shen Dillon

Organization: University of Illinois Urbana-Champaign

Title-Subject: [Filtered] Postdoc in in-situ nanomechanics

Message: Hello Everyone,

Our group is currently looking for a postdoctoral microscopist to work on in-situ nanomechanical
testing of tribofilms and solid-state lubricants. The position would require some travel to visit
with collaborators in the UK. If you know anyone who may be interested, please suggest that they
contact me. Thanks!

Best regards,
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Title-Subject: [Filtered] FOM-FIG "Lunch With a Manager" 2014 MM Hartford

Message: To My Colleagues,

The Facilities Operation & Management FIG is pleased to again host "Lunch With a Manager" forum at
the 2014 MM Meeting in Hartford. We invite participation by MSA and MAS student [Graduate and
Undergraduate] members interested in a career in Facility Management. The details and particulars
are listed below. If you would be so kind as to past this information along to any interested
students it would be appreciated. Please feel free to contact me with questions or comments.

Thanks!

Tom

An Invitation to MSA Student Members
Microscopy & Microanalysis 2014-Hartford
Â“Lunch with a ManagerÂ”

Sponsored by Facilities Operations &
Management*-Focused Interest Group

Considering a career managing a
Microscopy Facility, or just curious about what
it takes to run one?

The MSA FOM*-FIG is pleased to host a Lunch &
Roundtable forum with a panel of veteran
Facility Managers.

Discussion Topics:
--The Road Less Traveled: The Journey to becoming Â“The Lab ManagerÂ”.
--Â“Now What Do I Do?Â”: Dailey Challenges and Rookie Mistakes.

The Particulars:
Monday [4 Aug.]
Time: 12:15-1:30
Location: Capital 3 Rm. [Conn. Conv. Center]
Food & Drinks Provided**
**box lunch [vegetarian option]

Seating is limited to 20 participants on a first come
basis!

Deadline to register: 28 July

To Register:
Contact Tom Williams at the
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tomw-at-uidaho.edu
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Email: kalennon-at-hagerstowncc.edu
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Title-Subject: [Filtered] hand microtome

Message: Hi Fellow Microscopy Enthusiasts,

Do any of you have experience using a hand-held microtome? I have a colleague who is interested in
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Hand-held is not something I've ever used, and I'm a bit skeptical. I'd love to get her at least a
vibratome, but I'd bet they are out of our current price range.

Any advice?

Thanks once again for your help,
Kristen Lennon

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Congratulations on getting this far with it!

Donation SEMs often languish, because infrastructure and upkeep are far
more work than people realize, plus, they can't be put just anywhere due
to vibration issues. Pumps need to be vented. Sample prep isn't easy,
and often the necessary equipment for that is not donated along with it.

Protocols using gluteraldehyde and/or osmium tetroxide are especially
tricky, because a fume hood is rarely part of the donation. (You might
actually consider building something like this at your hackerspace.)

You've gotten much further than Noisebridge in San Francisco, where some
effort was expended to restore a tabletop SEM that was donated, but it
largely languished. Noisebridge is notoriously disorganized and in
decline, however.

You might consider a sliding scale fee for use of the SEM. If you
maintain the device at the level you should be, there's some definite
expense. SEM aperture plates and column apertures need to be replaced
annually, and that's a few hundred dollars each time. Also, oil changes
for the vacuum pumps should be done every 6 months, and the special
machine oil needed for that is kind of spendy. The vacuum pumps
themselves have a finite lifespan, and can be pretty expensive to
replace when they've reached the end of their life. And finding
replacements for parts for the SEM itself can be expensive if they
should break, if you can find them at all.

SEM can be a very good entry into science, not to mention a valuable
skill for high school students, if you can deal with the liability
issues. There are plenty of nonprofit educational efforts that
successfully negotiate this, however. And high schools aren't the
be-all-end-all for educational partnerships - consider community
colleges. Currently, I'm SEM tech at Ohlone College, which is one of the
few community colleges with an SEM and SEM classes. The other being
Delta College, which has an entire EM program. There may be a few other
examples. What I can tell you can find many people at that level who are
interested and would love the opportunity to work on such a device.
Consider working with biotech, biology, engineering, and other science
classes. It's possible to even offer an SEM class with the hackerspace
as an off-site - it might be possible to get some upkeep money for the
instrument this way as well.

Keep in touch about this - I'd be interested to know how your
hackerspace SEM project is going.

Cheers!
Peter G. Werner
Microscopy and Physical Science Lab Technician, Ohlone College
Lab Technician/Instructional Assistant, Microscopy, Merritt College
Acting President, San Francisco Microscopical Society
pwerner-at-ohlone.edu
(510) 979-7911 (office)
(415) 261-7114 (cell)

On 2014-05-13 06:00, rdpierce-at-pobox.com wrote:
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} First, I want to thank the members of this list for all the help
} keeping our donated SEM running for the past year. The advice and
} connections I've made have been extremely valuable.
}
} Now I'm asking a different kind of question. Some background:
}
} Pumping Station: One is a non-profit "hackerspace" or "makerspace"
} located in Chicago. We are an organization that provides members with
} access to all sorts of tools (woodworking, machine shop, electronics,
} welding, CNC, 3D printing, laser cutting, home brewing and cooking,
} sewing and machine knitting, etc.) but more than that, we have built a
} community of around 270 makers working on projects, offering classes,
} and offering plenty of informal mentoring and support for each other's
} projects.
}
} So, last year, we received a donation of a Leica S430. We believe we
} are the only hackerspace with a working SEM. I've been building up
} infrastructure for a microscopy lab around it. The scope works
} reliably (even if the vacuum is not as good as I'd like) with a
} repairable BSD and a working EDX, and we have a working sputter
} coater, critical point dryer, and ultrasonic cleaner. I'm finishing up
} a project to have bulk liquid nitrogen available, and it looks like I
} might have a source for a binocular inspection microscope for sample
} prep.
}
} Now I'm focusing on the "people" side of things, and what to do with
} all of this. We're in an unusual situation. Universities and labs have
} reasons to own a SEM. High schools with donated SEMs have curricula /
} after school programs. While our organization is educational, it isn't
} formal in the sense of a school, and a SEM is really an extra or
} bonus.
}
} I've put together a training program which is popular. People like to
} learn new tools. I've got more than 20 people authorized to use it on
} their own. Yet there has been very little use outside of the training;
} often people don't touch it afterwards. (In fairness, this is true of
} other tools too, e.g. welding, machining....)
}
} Part of it may be that I have not yet started offering sample prep
} classes / authorizations, although I've made it clear that I'm happy
} to prepare samples for people and load them in the chamber. So far
} only two people have taken me up on that. Admittedly, sample prep is
} an area I know little about, especially biological sample prep. (Did I
} mention that I'm pretty much self taught in all this?) The CPD was an
} impulse buy on eBay, before I realized we need to do glutaraldehyde
} fixing first which needs a fume hood to do safely, so I'm not sure how
} to handle wet biological samples. I'm left with a lot of questions.
} Like whether PLA or ABS plastics outgas at SEM vacuum levels, and
} whether they could be sputter coated and wouldn't melt under the beam.
} (We do a lot of 3D printing, and it would be interesting to compare
} these.) Or how to mount specimens with small particulates. (I made a
} trip to the Chernobyl exclusion zone, and I'd like to do EDX analysis
} on samples collected.)
}
} So I'm left puzzled. I think it is unprecedented to have an SEM
} available 24/7, free of charge with free training, to the general
} public in the middle of a major city, yet nobody seems to be taking
} advantage of it. (The general public would have to become members to
} use it, but membership is open to anyone 18 and older for $40 a month,
} and that includes access to all the other tools.)
}
} I'm not sure if people are intimidated by it, or if they have a hard
} time thinking of applications for it. I've considered having open "SEM
} Salons" where we would have a sample or project to work on as a group.
} But by and large, I'm looking for ideas of stuff to do, methods for
} sample prep, ways to do outreach to get people involved, etc.
}
} I'm also looking for groups to partner with. High schools are
} problematic at this time because we are still trying to nail down
} liability issues around minors. Collaboration with for-profit groups
} would be fine too, provided they did participate and give back to our
} community.
}
} And, of course, if anyone on this list is local to Chicago, we are
} looking for members to get involved with the work we are doing.
}
} Hackerspaces are a relatively new concept, and while we may be the
} only one now with a working SEM, I think that might change. I know a
} hackerspace in the Washington DC area is repairing a Cambridge
} Stereoscan 200. So I'm very much interested in best practices
} surrounding open science and public SEM access.
}
} Regards,
} Ryan Pierce
}
}
}
}
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For undergrad classes, we've always just used double-edged razor blades,
snapped in half. Students can either hold the tissue down with their
finger and slice across (like slicing an onion), or put the tissue between
pieces of potato or carrot to support it while cutting.

Students learn this easily, just need to start them on some big, easy to
cut tissue, e.g. Coleus stem or similar. Our 2nd year students learned all
their plant anatomy by doing a project sectioning differen parts of a
native seedling, some of these were quite woody! Reason for selecting
these plants is that there are no diagrams to copy from books, so they
have to look at their own tissue. (Cynical from experience.)

We have a couple of hand microtomes, they don't get used much. You need a
good old-fashioned large (beard) shaving blade to get good sections from
this, which is more dangerous than razor blades in my opinion.

A simple one is described here:
http://www.microscope-microscope.org/activities/school/microtome.htm

But see
http://prometheuswiki.publish.csiro.au/tiki-index.php?page=Making+hand+sect
ions+without+support+material for hand sectioning without support
material, and in the grey box on that page are some alternatives using
various support materials.

We still do most of our sectioning by hand...

cheers,
Rosemary

Dr Rosemary White
CSIRO Plant Industry
GPO Box 1600
Canberra, ACT 2601
Australia

T 61 2 6246 5475
F 61 2 6246 5334
E rosemary.white-at-csiro.au

On 15/05/14 10:17 AM, "microscopylistserver-noreply-at-microscopy.com"
{microscopylistserver-noreply-at-microscopy.com} wrote:

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From popular_medications9-at-margotcamille.com Thu May 15 04:36:46 2014
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Let me recommend single edge razors like GEM. They are sharp, have a safe back to press against, and are affordable. I thin and cross section a lot plastic and rubber components and they work fine.

Keep your fingers attached............
Frank

-----Original Message-----
X-from: Rosemary.White-at-csiro.au [mailto:Rosemary.White-at-csiro.au]
Sent: Wednesday, May 14, 2014 9:16 PM
To: Frank Karl

For undergrad classes, we've always just used double-edged razor blades,
snapped in half. Students can either hold the tissue down with their
finger and slice across (like slicing an onion), or put the tissue between
pieces of potato or carrot to support it while cutting.

Students learn this easily, just need to start them on some big, easy to
cut tissue, e.g. Coleus stem or similar. Our 2nd year students learned all
their plant anatomy by doing a project sectioning differen parts of a
native seedling, some of these were quite woody! Reason for selecting
these plants is that there are no diagrams to copy from books, so they
have to look at their own tissue. (Cynical from experience.)

We have a couple of hand microtomes, they don't get used much. You need a
good old-fashioned large (beard) shaving blade to get good sections from
this, which is more dangerous than razor blades in my opinion.

A simple one is described here:
http://www.microscope-microscope.org/activities/school/microtome.htm

But see
http://prometheuswiki.publish.csiro.au/tiki-index.php?page=Making+hand+sect
ions+without+support+material for hand sectioning without support
material, and in the grey box on that page are some alternatives using
various support materials.

We still do most of our sectioning by hand...

cheers,
Rosemary

Dr Rosemary White
CSIRO Plant Industry
GPO Box 1600
Canberra, ACT 2601
Australia

T 61 2 6246 5475
F 61 2 6246 5334
E rosemary.white-at-csiro.au

On 15/05/14 10:17 AM, "microscopylistserver-noreply-at-microscopy.com"
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Gary,

I am a satisfied user of an FEI 650 tungsten instrument and find it quite capable for general work. Realize, though, that my definition of 'general' is rarely exceeding 30K magnification on mainly metallic fractures and various inorganic unknowns. It is apparent to me while examining my samples, though, that if I were to need higher magnification, a FEG would be much preferred; the W instrument is current and/or aberration limited at both high mags, low voltages, as well as high count rate EDX mapping. Having had two variable pressure scopes (other vendors) before the 650, I don't understand your comment about the FEI variable pressure being different from others; you may be referring to the secondary electron imaging based on Danilatos' ESEM detectors using electron cloud amplification. That is true. Also, realize the beam spreading effects in variable pressure mode severely restrict high magnification resolution in any scope. I would much prefer a FEG instrument but couldn't justify the extra $250K.

Hope this helps,

Rick

Richard A. Ross
Sr. Metallurgist, Materials Engineering
Allison Transmission Inc.
Office: (317) 242-4880 - Fax (317) 242-7638
Richard.Ross-at-AllisonTransmission.com

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TESCAN USA Inc. is hiring three Field Service Engineers.

Positions are for Field Service Engineers based in the Northeast, Mid-Atlantic, and Southeast USA, to work out of their homes. The Field Service Engineer will be responsible for all aspects of customer service including routine maintenance, installations, emergency service, instrument troubleshooting and repair, customer training, and any necessary customer support associated with the position. The engineer will be required to be a self-starter and able to travel and work independently with minimal supervision. Good organizational skills and written and oral communication skills are a must. Maintaining good customer relations is essential.

Other requirements of the job include:

* Working knowledge of vacuum systems and components.
* Working knowledge of Electron Microscope electron optics and image detectors.
* Working knowledge of electronics and power supplies
* Solid mechanical abilities to analyze complex and intricate mechanical components.
* Computer skills of how PC's operate and Windows OS's from 2000 to Win 7
* Regularly required to crouch, crawl, stoop or kneel behind instrumentation in confined spaces
* Must have ability to see, feel and handle small objects and use hand tools
* Must have ability to lift 50 lbs.
* Submit timely service reports and travel expense reports
* If not US Citizen must have necessary authorization and documents to legally work in the USA
* Must have valid driver's license
* Must have valid Passport
* Have a car in good condition capable of traveling several hundred miles if necessary
* Must be able to pass a criminal and credit background check and drug testing.

TESCAN USA Inc. offers competitive salaries and benefits. Qualified candidates should send resume and cover letter along with salary requirements to info-at-tescan-usa.com and reference FSEMW. No phone calls will be accepted. Equal Opportunity Employer.

Colleen Leary
Strategic Marketing Manager
TESCAN USA
508 Thomson Park Dr.
Cranberry Twp., PA 16066
Phone: 724-772-7433
E-mail: cleary-at-tescan-usa.com

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When I teach sectioning, I like another poster, use a carrot or Styrofoam to hold the sample with single edge razors to section. For educational and basic functions it works fine for the plant materials we used. However, I do have a hand microtome that was made in Japan and it came with a big knife similar to an old barbers straight edge blade. It is designed to hold paraffin blocks for sectioning. However, I have never managed to get around to using it since the materials we research are for the most part dried powdered botanicals or drug/dietary ingredient materials. I picked it up in case. From what I understand, the product is not being made anymore but I am sure someone out there sells them. Unfrotunately, I can't provide any feedback on the student hand microtome since I have not used it but it is made well and looks solid.

Sincerely,

James Neal-Kababick
Director
Flora Research Laboratories
An FDA and DEA Registered & Inspected Laboratory
Fellow AOAC International
Adjunct Faculty Bastyr University
Botanical Medicine Department
1-541-472-0980 phone
jimk-at-floraresearch.com
www.floraresearch.com

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Email: kalennon-at-hagerstowncc.edu
Name: Kristen Lennon

Organization: Hagerstown Community College

Title-Subject: [Filtered] hand microtome

Message: Hi Fellow Microscopy Enthusiasts,

Do any of you have experience using a hand-held microtome? I have a colleague who is interested in purchasing one to use as part of an introduction to microscopy class for our undergraduates.
Hand-held is not something I've ever used, and I'm a bit skeptical. I'd love to get her at least a vibratome, but I'd bet they are out of our current price range.

Any advice?

Thanks once again for your help,
Kristen Lennon

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Greetings microscopical friends,

I have the prospect of doing TEM on some skin samples that have been embedded in Optimal Cutting Temperature medium and (presumably) plunge frozen. Knowing that skin can be tricky anyway for TEM fixation, and that plunge freezing 4 mm wide samples might result in horrible ice damage, I am feeling a bit apprehensive about what to expect.

I am grateful for any advice or experiences with OCT and TEM (even just a short reply).

1. Is it so likely to fail that it should not be attempted?
2. If it can be attempted, what should I expect (and describe) the likely quality as being?
3. Is there a protocol or any tricks (e.g., use edge or middle of 4 mm sample only, freeze substitution rather than thaw and fix etc.)?

Thanks
Duane

AgResearch Limited | Lincoln Research Centre | New Zealand
Dr Duane P Harland, Senior Scientist
T +64 3 321 8710 E duane.harland-at-agresearch.co.nz

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Sorry to say so but I would guess your option 1 that it is "so likely to fail that it should not be attempted" is the right prediction. I have done a lot of quick-freezing with metal mirrors and even those specimens often fail. I have seen a ton of poorly frozen tissue and it isn't worth looking at. TEM is a great technique but it is too tedious and time-consuming to try unless you at least try to do the fixation in an optimal way. Plunge freezing is unlikely to be worth the effort. Tom

Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
phillipst-at-missouri.edu

http://biology.missouri.edu/people/?person=88
http://www.biotech.missouri.edu/mcc/

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To: Phillips, Thomas E.

Greetings microscopical friends,

I have the prospect of doing TEM on some skin samples that have been embedded in Optimal Cutting Temperature medium and (presumably) plunge frozen. Knowing that skin can be tricky anyway for TEM fixation, and that plunge freezing 4 mm wide samples might result in horrible ice damage, I am feeling a bit apprehensive about what to expect.

I am grateful for any advice or experiences with OCT and TEM (even just a short reply).

1. Is it so likely to fail that it should not be attempted?
2. If it can be attempted, what should I expect (and describe) the likely quality as being?
3. Is there a protocol or any tricks (e.g., use edge or middle of 4 mm sample only, freeze substitution rather than thaw and fix etc.)?

Thanks
Duane

AgResearch Limited | Lincoln Research Centre | New Zealand Dr Duane P Harland, Senior Scientist T +64 3 321 8710 E duane.harland-at-agresearch.co.nz

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18, 31 -- From PhillipsT-at-missouri.edu Thu May 15 17:56:33 2014
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From meds_popular10-at-prtcom.ru Thu May 15 21:38:08 2014
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Good morning,
dear Kristen and all,

perhaps the pdf via link http://www.modernmicroscopy.com/article_pix/040810_quarterblades/quarterblades.pdf will
(TEETSOV Anna: Quarter-Razor blade for hand-sectioning) will contribute a little bit.

I recall a very old publication (technical note, 1960/70ies?) on a hand-made {serial cutting} instrument, where,
several (full sized) razor-blades were mounted into a â€œholder constructionâ€ with at least two leading pins (as in wet shavers for men), up to 5 or 6 razor blades slipped over using kind of (really thin, according to the thickness of slices wanted) spacer/distance rings in between each razor blade, fixed at both ends and equipped with a handle. The â€œinstrumentâ€ was fabricated either in a shop or was built as handicraft work. This way one would be able to â€œsectionâ€ slices/slabs (like â€œmicro-grossingâ€ with a defined thickness [equidistant] from the original big specimen [from abalone to zysts (â˜º)])
Unfortunately I canâ€™t find the article/sketch of that instrument, nevertheless I think this is worth to be mentioned.

Very best regards,

Wolfgang
SALZBURG, AUSTRIA

Secretary of SCUR
PLEASE VISIT: www.scur.org
for the Program of 41st Annual SCUR(Society for Cutaneous Ultrastructure Research) -Meeting, 5-7 June 2104 in Monte Porzio Catone, ROME, Italy

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Von: microscopylistserver-noreply-at-microscopy.com [mailto:microscopylistserver-noreply-at-microscopy.com]
Gesendet: Donnerstag, 15. Mai 2014 02:07
An: MuÃŸ Wolfgang
Betreff: [Microscopy] Hand-held microtome

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Hi Fellow Microscopy Enthusiasts,
Do any of you have experience using a hand-held microtome? I have a colleague who is interested in purchasing one to use as part of an introduction to microscopy class for our undergraduates.
Hand-held is not something I've ever used, and I'm a bit skeptical. I'd love to get her at least a vibratome, but I'd bet they are out of our current price range.

Any advice?

Thanks once again for your help,
Kristen Lennon

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Title-Subject: [Filtered] Postdoc in in-situ fluid cell imaging and
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Message: Greetings Everyone:

Our group is currently looking for a postdoctoral microscopist to work
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Dear All: I am helping to start out the cryo-tem in my Lab and is planning to train new users who need to use this technique. While generally i am familiar with the procedures in cryo-plunging, sample transfer from tem-box to Gatan cryo-holder in the cryo-workstation, i cannot perform such procedure if i am wearing a pair cryo-gloves becausae the gloves are thick and it is impossible use them to hold the tweezers and to pick up the copper grid in the pool of liquid nitrogen/liquid ethane.That would mean that my hands are not protected from the LN2 and this can be risky. My Insitute is very particular on safety and i need to conduct a Risk Assessment on this procedures. May i ask how safety is implemented during these procedures?

Another enquiry would be on the technical aspect during sample transferring. I notice that if i have to transfer the grid box from the dewar (during storage) to the Gatan workstation, the grid box will have to expose to the atmosphere where a rise in temperature and icing are expected. This can be minimized when i quickly move the grid box into the workstation. However, i find that it takes a longer time to transfer the gridbox from the workstation back to the conical tube or into the dewar. Will there a better solution to reduce the time for the gridbox to expose to the atmosphere and humidity?

There is a Cryo Grid Box Handling Tool given by Gatan, is it very useful in minimize temperature rise and humidity when transfering the gridbox from the Gatan workstation to the storage dewar/conical tube?

Thank you very much!

Cheers,
Yee Yan, Tay
FACTS Lab
NTU, Singapore

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Hi All

Here in the UK we have a health and safety clause called "best practicable
means", this basically says if you cannot perform the action you may take
another route. To handle cryo related components with sufficient dexterity
is often made impossible using what would be called suitable gloves. In
which case we would fall back on the clause, what happens in other
countries?

I am responsible for making health and safety checks on car race tracks, so
you may imagine the complex areas that have to be covered by the same
clause.

Good luck

Steve

Steve Chapman FRMS
Protrain for Consultancy and Courses in Electron Microscopy
Mobile +44 (0) 7711 606967 Fax +44 (0)1280 814007
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-----Original Message-----
X-from: rongchigram79-at-yahoo.com.sg [mailto:rongchigram79-at-yahoo.com.sg]
Sent: 18 May 2014 06:12
To: protrain-at-emcourses.com

Dear All,

As the deadline for early registration for this yearâ€™s Microscopy and
Microanalysis Meeting (Aug. 3-7 2014 in Hartford, CT) is coming
closer, I would like to make you aware of our Sunday Short Course on
3D electron microscopy (description below). Please forward this also
to your colleagues who may be interested in this topic.
This course is ideal for people who need a comprehensive introduction
or a refresher in 3D reconstruction methods and is suitable for
biologists and material scientists.
If you have already registered for the meeting you can add the course
registration as â€œextra item onlyâ€.

X12 3D Electron Microscopy of Macromolecular Assemblies
Teresa Ruiz, Michael Radermacher, Edward Morris
8:30 AM to 5:00 PM on Sunday, August 3.

â€¢ Sample preparation: deep stain, vitreous ice
â€¢ Imaging conditions, low-dose imaging, tilt-pair data collection
â€¢ Alignment techniques and multivariate statistical analysis
â€¢ 3D reconstruction methods
â€¢ X-ray structure docking
â€¢ Techniques described have applications in both biological and
material science

This short course will provide a comprehensive description of the
methods used for 3D structure determination from electron micrographs
of macromolecular complexes or weakly scattering specimens available
in multiple copies. Specimen-preparation techniques for single
particles (deep stain, vitreous ice) will be presented, followed by
selection of optimal imaging conditions, including low-dose imaging.
Next, a detailed explanation of image-processing techniques, with
special emphasis on the random-conical reconstruction technique, will
be presented. Finally, structure interpretation and docking of X-ray
structures to 3D EM densities will be demonstrated. The techniques
described could be applied to both biological and materials science
specimens.

--
Michael Radermacher, Ph.D., Prof., FMSA
University of Vermont
Dept. Molecular Physiology and Biophysics
HSRF 120 / 149 Beaumont Ave
Burlington, VT 05405
USA

Tel: + 802 656-4834
Fax: + 802 656-0747

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Hello cryoSEM users.

I'm interested in hearing about your experiences with various cryoSEM "glues" for sticking down samples to holders prior to freezing, including pitfalls, advantages, disadvantages, etc. Also, for live insect or other invertebrate samples, what are your suggestions for immobilizing them so they can be stuck to the glue prior to freezing. Thanks for your advice.

Marie

Dr. Marie E. Cantino
Associate Professor of Physiology and Neurobiology
Director, Electron Microscopy Laboratory
University of Connecticut, Unit 3242
Phone: 860-486-3588
Fax: 860-486-6369

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Hi

I was involved with the development of one of the first SEM cryo systems and
since that time have always used Tissue Tec as my adhesive of choice.

Regards

Steve

Steve Chapman FRMS
Protrain for Consultancy and Courses in Electron Microscopy
Mobile +44 (0) 7711 606967 Fax +44 (0)1280 814007
www.emcourses.com

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-----Original Message-----
X-from: marie.cantino-at-uconn.edu [mailto:marie.cantino-at-uconn.edu]
Sent: 19 May 2014 19:34
To: protrain-at-emcourses.com

We used to use TissueTek, but switched to Instrumedics Cryo-Gel because
it's so much easier to handle. It's a gel, so doesn't run all over the
place before being frozen. We use it a lot to arrange our samples
precisely before cryostat sectioning. If there's a gel form of TissueTek
I'm sure that'd be fine too.

cheers,
Rosemary

Dr Rosemary White
CSIRO Plant Industry
GPO Box 1600
Canberra, ACT 2601
Australia

T 61 2 6246 5475
F 61 2 6246 5334
E rosemary.white-at-csiro.au

On 20/05/14 4:43 AM, "marie.cantino-at-uconn.edu" {marie.cantino-at-uconn.edu}
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I also use Tissue Tek but colloidal graphite also works well.

Regards
Mark

Dr Mark A.E. Auty
National Food Imaging Centre
Food Chemistry & Technology Department
Teagasc Food Research Centre
Moorepark, Fermoy, Co. Cork
Ireland

Land line: +353 (0) 25 42442
Mobile: +353 (0) 87 0663732
Fax: +353 (0) 25 42221
Email: mark.auty-at-teagasc.ie

-----Original Message-----
X-from: Rosemary.White-at-csiro.au [mailto:Rosemary.White-at-csiro.au]
Sent: 19 May 2014 23:56
To: Mark Auty

We used to use TissueTek, but switched to Instrumedics Cryo-Gel because it's so much easier to handle. It's a gel, so doesn't run all over the place before being frozen. We use it a lot to arrange our samples precisely before cryostat sectioning. If there's a gel form of TissueTek I'm sure that'd be fine too.

cheers,
Rosemary

Dr Rosemary White
CSIRO Plant Industry
GPO Box 1600
Canberra, ACT 2601
Australia

T 61 2 6246 5475
F 61 2 6246 5334
E rosemary.white-at-csiro.au

On 20/05/14 4:43 AM, "marie.cantino-at-uconn.edu" {marie.cantino-at-uconn.edu}
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From pharmacy-popular2-at-goldberglawcenter.com Tue May 20 07:45:24 2014
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Dear Jen, Steve , Guenter and Paul,

Thank you very much for your suggestions and recommendations. A consensus would be to avoid wearing gloves which does makes sense because it would be extremely dangerous if LN2 is trapped. On the other hand it would be interesting for me to check with my safety office if 'the best practicable means' exist in my country.

Guenter and Jen, if you didn't mind, could you send me the link to access the safety presentation as well as the picture for the modifid Nalgene bottle?

Cheers,
Yee Yan
FACTS Lab
NTU

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Contents Retrieved from Microscopy Listserver Archives
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Fellow Microscopists,

The Midwest Microscopy and Microanalysis Society will hold a meeting in conjunction with the University of Wisconsin-Madison on June 6, 2014. The program title is "In Situ Microscopy", and events also include a Facility Open House at the UW-Madison Materials Science Center on Thursday, June 5th. The open house schedule, a list of Friday's speakers and registration information can be found on our website Meetings page:

www.midwestmicroscopy.org

Please contact Ms. Kristy Wendt, kwendt3-at-wisc.edu, for registration and vendor information. We look forward to seeing you there!

Best regards,

Elaine Schumacher
M3S Program Coordinator

*********************************************************************
Elaine F. 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

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

==============================Original Headers==============================
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Living and working in the desert SW, we have a number of researchers here looking at the effects of dust on the body (asthma, mine tailings, etc). I was recently asked if we could look at the dust using light microscopy.

Since I am a biologist by training, I have little experience with this kind of material. Can someone recommend some reference/reading material? I am probably not ready (yet) for a McCrone Research Institute course, but I am aware of their existence.

Thanks!
Doug

^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^
Douglas W. Cromey, M.S. - Associate Scientific Investigator
Dept. of Cellular & Molecular Medicine, University of Arizona
1501 N. Campbell Ave, Tucson, AZ 85724-5044 USA

office: AHSC 4212 email: Cromey-at-Arizona.edu
voice: 520-626-2824 fax: 520-626-2097

http://swehsc.pharmacy.arizona.edu/micro
Home of: "Microscopy and Imaging Resources on the WWW"

UA Microscopy Alliance - http://microscopy.arizona.edu

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9, 31 -- From dcromey-at-email.arizona.edu Tue May 20 09:51:07 2014
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9, 31 -- From: "Cromey, Douglas W - (dcromey)" {dcromey-at-email.arizona.edu}
9, 31 -- To: "Microscopy listserv (Microscopy-at-Microscopy.Com)"
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9, 31 -- Subject: LM of dust
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You can mount this material in a product called Cargille Meltmount 1.662 (other RI available too) which you can get from McCrone. It is available from McCrone and it is easy to use. There are instructions you can download or that will come with the medium. You can also do a wet mount in glycerol--water but the other mount is a "permanent" mount in that unless you keep the slide on edge in a very hot area, it will remain. The nice thing about meltmount is that you can reheat the slide and rotate the fibers or other specimen in the mountant for PLM. There is a great book that might be of value to you called Pharmaceutical Microscopy. Another fun book it the color Atals and Manual of Microscopy for Criminalists, Chemists and Conservators by CRC Press. This is kind of an interesting collection of techniques, applications and illustrations (lots of photos). There is a chapter on dust.

One other thing is that you can register on the McCrone site for free access to the particle atlas and view images of dust with details about the mounting technique and various images from optical light microscopy to EM. You can also purchase reference slides from McCrone or Wards of dust mounted in a medium like Meltmount.

There is a lot of interesting things to find in dust. A good way to practice is to collect some floor sweepings from the garage, kitchen or a public area for examination. I hope that this helps.

Sincerely,

James Neal-Kababick
Director
Flora Research Laboratories
An FDA and DEA Registered & Inspected Laboratory
Fellow AOAC International
Adjunct Faculty Bastyr University
Botanical Medicine Department
1-541-472-0980 phone
jimk-at-floraresearch.com
www.floraresearch.com

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-----Original Message-----
X-from: dcromey-at-email.arizona.edu [mailto:dcromey-at-email.arizona.edu]
Sent: Tuesday, May 20, 2014 8:14 AM
To: James Neal-Kababick

Living and working in the desert SW, we have a number of researchers here looking at the effects of dust on the body (asthma, mine tailings, etc). I was recently asked if we could look at the dust using light microscopy.

Since I am a biologist by training, I have little experience with this kind of material. Can someone recommend some reference/reading material? I am probably not ready (yet) for a McCrone Research Institute course, but I am aware of their existence.

Thanks!
Doug

^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^
Douglas W. Cromey, M.S. - Associate Scientific Investigator Dept. of Cellular & Molecular Medicine, University of Arizona
1501 N. Campbell Ave, Tucson, AZ 85724-5044 USA

office: AHSC 4212 email: Cromey-at-Arizona.edu
voice: 520-626-2824 fax: 520-626-2097

http://swehsc.pharmacy.arizona.edu/micro
Home of: "Microscopy and Imaging Resources on the WWW"

UA Microscopy Alliance - http://microscopy.arizona.edu

==============================Original Headers==============================
23, 31 -- From jimk-at-floraresearch.com Tue May 20 11:36:36 2014
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On May 17, 2014, at 10:21 PM, rongchigram79-at-yahoo.com.sg wrote:

} Dear All: I am helping to start out the cryo-tem in my Lab and is
} planning to train new users who need to use this technique. While
} generally i am familiar with the procedures in cryo-plunging, sample
} transfer from tem-box to Gatan cryo-holder in the cryo-workstation,
} i cannot perform such procedure if i am wearing a pair cryo-gloves
} becausae the gloves are thick and it is impossible use them to hold
} the tweezers and to pick up the copper grid in the pool of liquid
} nitrogen/liquid ethane.That would mean that my hands are not
} protected from the LN2 and this can be risky. My Insitute is very
} particular on safety and i need to conduct a Risk Assessment on this
} procedures. May i ask how safety is implemented during these
} procedures?
}
}
} Another enquiry would be on the technical aspect during sample
} transferring. I notice that if i have to transfer the grid box from
} the dewar (during storage) to the Gatan workstation, the grid box
} will have to expose to the atmosphere where a rise in temperature
} and icing are expected. This can be minimized when i quickly move
} the grid box into the workstation. However, i find that it takes a
} longer time to transfer the gridbox from the workstation back to the
} conical tube or into the dewar. Will there a better solution to
} reduce the time for the gridbox to expose to the atmosphere and
} humidity?
}
} There is a Cryo Grid Box Handling Tool given by Gatan, is it very
} useful in minimize temperature rise and humidity when transfering
} the gridbox from the Gatan workstation to the storage dewar/conical
} tube?
}
} Thank you very much!
}
} Cheers,
} Yee Yan, Tay

Dear Tay,
As others have said, trying to use cryo-gloves is bad practice. In
fact, it is much more likely that you could be harmed trying to do the
fine manipulations required while wearing these than without gloves.
Liquid nitrogen (LN2) will not harm the skin unless it stays in
contact for an extended time, but the same is not true of liquid
ethane or propane. Occasionally small drops of this cryogen can be
produced during the plunging process, and if they contact your skin
you will see initially a white patch, typically ~1 mm in diameter, on
the skin, which becomes a small blister. This is generally only an
annoyance. A certain level of skill is required to manipulate the
grids with cooled tweezers. If one leaves them cooling too long, the
upper part can get too cold, but too little cooling of the tips can
heat the grid. I have taught people to move quickly, but not to rush,
when performing the necessary actions.
When transferring a grid box, keep it upright so that some LN2 is
present in the slots holding the grids. I usually use a large
tweezer, ~20 cm long with ~2 mm wide tips, to grab the box either from
the tube or station. Once again move quickly, but don't rush, and
drop the box into the station or tube. If you want to remove a box
from a tube containing other boxes, I use an intermediate holder. A
styrofoam box ~10 cm x 10 cm x 3 cm deep is good for this. Dump out
all the boxes from the tube, find the one you want, then put the
others back. I have not had any problems with ice on either the grid
boxes or the grids themselves using these procedures. Bear in mind
that there is cold N2 vapor above the LN2, so anything in that layer
will stay cold, but too rapid movement can mix the cold layer with
room air; another reason not to rush your movements.
I have found the old style covers, disks with a slot that can be
rotated to uncover a slot or to keep them all covered, work better
than the newer style "witches hat" covers. The newer covers often
drag a grid out of its slot as they are unscrewed, and this is
particularly problematic if the cover is opaque. It is also the case
that the boxes with the disk covers are more compact, so more of them
will fit into a tube. Our lab had a 13 mm punch, and we bought 30 cm
x 30 cm sheets of an appropriate plastic--it has to be OK at LN2
temperature, and it is best if it is transparent. We punched out as
many disks as we would need, drilled a central hole for a screw and
another for the inner end of the slot, then made the rest of the slot
with a Dremel tool. I prefer using steel screws to plastic ones,
since I have broken a few plastic ones trying to loosen them.
Yours,
Bill

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Greetings listers,

A colleague is looking to purchase a piezo-cooled stage for light
microscopy, +/- 0.1C, range 2-35C. Any experiences or advice would be
most welcome. Please contact me off list.

Thanks in advance,

Jim

--
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Digital Microscopy Facility
Mount Allison University
63B York St.
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CANADA

phone: 506-364-2519
fax: 506-364-2505
email: jehrman-at-mta.ca
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Sorry, I had a neuron crossed somewhere. Meant to be Peltier-cooled. Too
late in the day.

Jim

Greetings listers,

A colleague is looking to purchase a piezo-cooled stage for light
microscopy, +/- 0.1C, range 2-35C. Any experiences or advice would be
most welcome. Please contact me off list.

Thanks in advance,

Jim

--
James M. Ehrman
Digital Microscopy Facility
Mount Allison University
63B York St.
Sackville, NB E4L 1G7
CANADA

phone: 506-364-2519
fax: 506-364-2505
email: jehrman-at-mta.ca
www: http://www.mta.ca/dmf

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Title-Subject: [Filtered] Confocal microscopes

Message: Does anyone have positive or negative experience with Bio Rad
confocal microscopes and the E800 Nikon microscope? We are looking to
buy a used confocal microscope, and we are trying to decide on the Bio
Rad system.

Questions include
ease of use, service, student friendly, etc?

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Title-Subject: [Filtered] How to diagnose 'no filament current' jeol 6100

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From pharmacy-express11-at-wfol.com Fri May 23 02:38:50 2014
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Dear Joe,

We have experience with a wide variety of optical microscopes and
like many of the. Nikon Eclipse is a great stand.... definitely user
friendly. Also, one which, to the best of our knowledge, has a good
track record for stability and flexibility. Nikon has been
especially energetic in its lens development program so if it doesn't
come with all the objectives/eyepieces/condensers that you need for
your particular experiments, there is a wide selection to choose from.

Re: BioRad
You did not specify which model. When BioRad was in its hey-day, I
was working with Sarastro, a main competitor. The biggest issue for
you to check out is the alignment procedure. If it is one of the
earlier models, the light path may be complex... OK if you have
someone who can manage it, but not for novices. Also, see if you can
run a sample. Look for over-exposure. The only other issue to check
out is the nature of the pinholes. If you are doing co-localization
studies, you want to make sure that there is some mechanism for
setting the pinholes in front of each the"red" and "green" channels
exactly the same. Otherwise, you end up taking information from
different depths of field in each channel.

Hope this was helpful .. and Good Hunting!
Barbara Foster, President & Chief Consultant
Microscopy/Microscopy Education*
www.MicroscopyEducation.com

*A subsidiary of The Microscopy & Imaging Place, Inc.
7101 Royal Glen Trail, Suite A
McKinney, TX 75070
P: 972-924-5310
F: 214-592-0277

MME is currently scheduling courses for now and through Fall 2014.
Call us today for a free training evaluation.

At 02:15 PM 5/23/2014, you wrote:

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Good afternoon all,

I have purchased my first bag of sapphire discs to grow cells on for HPF. I am looking for advice on how to handle them. Do I need to clean them before using them? If so, how?

Then I would like to know if there is a trick for how to carbon coat them so that you get them in and out of the coater without losing them.

Does anyone have a trick for how to label or mark the discs so that you know what side the cells were grown on in case you drop the disc before it is frozen?

Thanks in advance for all suggestions.

Chris
Christine A. Brantner, PhD [C] NIH
NHLBI Electron Microscopy Core Facility
National Institutes of Health
14 Service Road West
Building 14E Room 111B MSC5570
Bethesda, MD 20892-5570

301-496-4711
chris.brantner-at-nih.gov {mailto:chris.brantner-at-nih.gov}

http://www.nhlbi.nih.gov/research/intramural/researchers/core/electron-microscopy-core/index.html

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Hi Rod
If you already have checked for broken filaments without improvement, it may be tricky and possibly expensive.
1. check all fuses,
2. check the HV cable using a insulation meter (megaohmmeter capable of 1000V) between both ends of the cable. It should read values below 1 Ohm.
3. If the cable is ok the HV-tank may be defect (happened to me after moving a J35c 100 m. Or the Load current control unit if you are lucky.

1&2 you can do yourselves, but you will at least need telephone advice or email advice for no 3 & 4 from a service technician.
Erik

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Title-Subject: [Filtered] embedding

Message: Anyone have any luck embedding samples for TEM- good or bad -
that have been sitting in ethanol for 2-3 weeks.

Cheers,
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Email: joseph.uknalis-at-ars.usda.gov
Name: Joe Uknalis

Title-Subject: [Filtered] Vacuum interlock fail on Edwards Scancoat 6

Message: When I turn on the vacuum pump the interlock light no longer
comes on the sputter coater (I still hear a click like a valve is
closing or such).

Is there a fix for this? Open up and blow off accumulated dust on a
sensor?? Or is it replaceable?

It worked fine up till this point, only purchased in 1999.

thanks

Joe

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From drugs-canadian8-at-chello.pl Sat May 24 12:38:53 2014
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This may be too simplistic but could the bulb be burnt out?

zaluzec-at-microscopy.com wrote:
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} Email: joseph.uknalis-at-ars.usda.gov
} Name: Joe Uknalis
}
} Title-Subject: [Filtered] Vacuum interlock fail on Edwards Scancoat 6
}
} Message: When I turn on the vacuum pump the interlock light no longer
} comes on the sputter coater (I still hear a click like a valve is
} closing or such).
}
} Is there a fix for this? Open up and blow off accumulated dust on a
} sensor?? Or is it replaceable?
}
} It worked fine up till this point, only purchased in 1999.
}
} thanks
}
} Joe
}
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From canadian-drugstore15-at-mts-nn.ru Sun May 25 08:29:15 2014
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Email: pa_grendys-at-fischione.com
Name: Paula Grendys

Organization: E. A. Fischione Instruments, Inc.

Title-Subject: [Filtered] Fischione Instruments is hiring Field Service Engineer - Japan

Message: Field Service Engineer, Japan (in-country work location)

E. A. Fischione Instruments, Inc., located in Export, Pennsylvania, USA, is a world leader in
microscopy and nanotechnology applications for the scientific community. In support of our ongoing
international sales growth, we are seeking candidates for the position of Field Service
Engineer-Japan to provide service to our clients located in Asia.

Located in-country Japan, and having the ability to work from your personal residence, this position
requires strong self-initiative and the ability to work with little direct supervision. Relocation
to Japan is available for qualified candidates.

In this key, customer-facing position, the Field Service Engineer-Japan is responsible for:

Â• Installing new equipment, executing product upgrades, and responding to service calls at client
locations
Â• Performing highly complex service and testing processes on miniature components, precision
equipments, and complex instrumentation
Â• Documenting service work, recording and analyzing test data, and preparing service reports
Â• Promoting customer loyalty through proactive, professional communication.
Â• Using company-supplied laptop to access technical and customer information
Â• Placing inventory and parts orders
Â• Providing information and feedback to Engineering and Production teams for problem identification
& resolution
Â• Providing comprehensive technical support by phone and email and promoting customer satisfaction
through exemplary customer service orientation
Â• Being "on-call" outside of normal working hours to provide emergency service.

Qualifications
Â• Requires a degree in electronics, electromechanics, or a related discipline.
Â• Must be able to write and speak English and Japanese.
Â• Minimum of 5 yearsÂ’ experience in a technical service capacity; prior field service responsibility
for complex, precision, electronic and/or electromechanical devices is preferred.
Â• Must possess the ability to interpret complex technical drawings, electronic schematics to the
component level, and wiring diagrams.
Â• Familiarity of standard vacuum practices and high-voltage technology is a plus.
Â• Needs to take initiative and work independently, with a focus on results, quality, and customer
service.
Â• Understanding of computer networking is beneficial.
Â• Excellent verbal and written communication skills are required.
Â• Good organizational habits and outstanding attention to detail are essential.
Â• Must have a valid driverÂ’s license and acceptable driving record.
Â• Must be willing to travel up to 50% of the time, in Japan and to nearby regions in Asia.
Â• Must be able to pass government background checks in order to be admitted to some customer facilities.

Working conditions and physical requirements
Â• Candidate will spend the majority of time in laboratory surroundings.
Â• This job occasionally requires exerting up to 50 kilograms of force.
Â• Must be able to sit, bend, kneel, and stoop; work may take place seated at a work station, on the
floor, or on a step stool or ladder.

Please visit our Web site to learn more about our company at http://www.fischione.com
See Career Opportunities for this position and link to apply online.

EOE

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Email: pa_grendys-at-fischione.com
Name: Paula Grendys

Organization: E. A. Fischione Instruments, Inc.

Title-Subject: [Filtered] Fischione Instruments is hiring Field Service Engineer - US

Message: Field Service Engineer, U.S. - PA

E. A. Fischione Instruments, Inc., located in Export, Pennsylvania, is a world leader in microscopy
and nanotechnology applications for the scientific community.

In support of our ongoing sales growth, we are seeking candidates for the position of Field Service
Engineer-US.
This position requires strong self-initiative and the ability to work with little direct supervision.

In this key, customer-facing position, the Field Service Engineer-US is responsible for:
Â• Scheduling and performing maintenance on scientific equipment at customer locations in accordance
with service contract and company procedures and standards. Ensuring equipment meets performance
specifications.
Â• Evaluating, diagnosing, and repairing malfunctioning equipment in response to service calls.
Â• Performing service on both ultra-precision subassemblies and complex instrumentation.
Â• Working to extreme tolerances and both recognizing and taking the action necessary to achieve
ultra-precise operation.
Â• Performing highly complex service and testing processes, sometimes with miniature components.
Â• Installing new equipment, executing product upgrades, and providing technical support.
Â• Promoting customer satisfaction through proactive, professional communication.
Â• Using company-supplied laptop to access technical and customer information.
Â• Documenting service work, recording and analyzing test data, and preparing service reports.
Â• Placing inventory and parts orders.
Â• Providing comprehensive technical support by phone and email.
Â• Being "on-call" outside of normal working hours to provide emergency service.

Qualifications
Â• Requires an associate degree in electronics, electromechanics, or a related discipline.
Â• Minimum of 5 years of experience in a technical service capacity; prior field service
responsibility for complex, precision, electronic and/or electromechanical devices is preferred.
Â• Must be able to interpret complex technical drawings, electronic schematics to the component
level, and wiring diagrams.
Â• Familiarity of standard vacuum practice and high-voltage technology is a plus.
Â• Needs to take initiative and work independently with a focus on results, quality, and customer
service.
Â• Understanding of computer networking is beneficial.
Â• Excellent verbal and written communication skills are required.
Â• Good organizational habits and outstanding attention to detail are essential.
Â• Must have a valid driverÂ’s license and acceptable driving record.
Â• Must be willing to travel up to 50% of the time, both domestically and internationally.
Â• Must be able to pass federal background checks in order to be admitted to some customer facilities.

Working conditions and physical requirements
Â• Candidate will spend the majority of time in laboratory surroundings.
Â• This job occasionally requires exerting up to 110 pounds of force.
Â• Must be able to sit, bend, kneel, and stoop; work may take place seated at a work station, on the
floor, or on a step stool or ladder.

Please visit our Web site to learn more about our company at http://www.fischione.com
See Career Opportunities for this position and link to apply online.

EOE

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From popular-meds5-at-airtelbroadband.in Mon May 26 11:33:42 2014
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Email: alexandre.bastien-at-fsaa.ulaval.ca
Name: Alexandre Bastien

Organization: Université Laval

Title-Subject: [Filtered] Brightfield image analysis

Message: Hi list,

I got a large number of BF image that needed to be analyzed in a full automatic way. Our images
covers a strongly different from one another. Here are some samples:

http://i58.tinypic.com/14e750n.jpg
http://i57.tinypic.com/313fzbc.jpg
http://i57.tinypic.com/2d9wlj.jpg

I need to do a cell segmentation, evaluate cell area, interstitial area, nucleus position relative
to the closest cell border, and maybe more.

I've tried severeal algorithm in Matlab and ImageJ, but none of them seems to cover all of our image
types, and analysis needs.

Do you know a software that can analyze this automatically ?

Many thanks.

Alexandre

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Dear Alexandre,

The main thing you need to do is capture the images appropriately so they
can be analysed. It's always more efficient to put the effort into good
image capture. Then the software can deal with the images quite easily.
But if you have to work with images like the ones in your email rather
than re-take them, you may have to do quite a bit of non-automated work,
unless you have access to rather sophisticated analysis software.

I would first do shading correction and then you may be able to play
around with segmentation based on hue/colour, and detection of outlines by
intensity gradients. Others on this list with much more analysis
experience may be able to provide more detailed advice.

good luck,
Rosemary White

Dr Rosemary White
CSIRO Plant Industry
GPO Box 1600
Canberra, ACT 2601
Australia

T 61 2 6246 5475
F 61 2 6246 5334
E rosemary.white-at-csiro.au

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From drugstore-canadian14-at-abt-inc.com Tue May 27 06:15:45 2014
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Greetings,

Although I have not fully automated the protocol, I have used CellSens (which I am new to using) which I got with my microscope and it makes analysis of distances and areas very simple and rapid. What used to take me hours to do with my ocular micrometer is done in minutes now. I have been able to rapidly determine mean areas for fungal spores as well as measurement of mean spore width and height and take it into excel with a few mouse clicks. I would say that I have to agree with Dr. White in that image quality will impact the ease of use of any package as well as the ability to manually measure with a program (versus fully automated analysis of images).

Sincerely,

James Neal-Kababick
Director
Flora Research Laboratories
An FDA and DEA Registered & Inspected Laboratory
Fellow AOAC International
Adjunct Faculty Bastyr University
Botanical Medicine Department
1-541-472-0980 phone
jimk-at-floraresearch.com
www.floraresearch.com

CAUTION:
This e-mail and its accompanying attachments, if any, may contain information that is confidential and subject to legal privilege. If you are not the intended recipient, you are notified that any use, dissemination, distribution or copying of this message is prohibited. If you have received this message in error, please notify Flora Research Laboratories immediately and destroy all copies of this message and the accompanying attachments. Thank you.

-----Original Message-----
X-from: Rosemary.White-at-csiro.au [mailto:Rosemary.White-at-csiro.au]
Sent: Tuesday, May 27, 2014 12:36 AM
To: James Neal-Kababick

Dear Alexandre,

The main thing you need to do is capture the images appropriately so they can be analysed. It's always more efficient to put the effort into good image capture. Then the software can deal with the images quite easily.
But if you have to work with images like the ones in your email rather than re-take them, you may have to do quite a bit of non-automated work, unless you have access to rather sophisticated analysis software.

I would first do shading correction and then you may be able to play around with segmentation based on hue/colour, and detection of outlines by intensity gradients. Others on this list with much more analysis experience may be able to provide more detailed advice.

good luck,
Rosemary White

Dr Rosemary White
CSIRO Plant Industry
GPO Box 1600
Canberra, ACT 2601
Australia

T 61 2 6246 5475
F 61 2 6246 5334
E rosemary.white-at-csiro.au

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Via Allen Glazner at UNC

I have resurrected an ancient Buehler Vibromet polisher (model 67-1517) and would like to use it to
put a final polish on probe samples. I have been unable to find cloths or abrasive pads for it (12-inch model)
and wonder if anyone has experience with this polisher.

Nick

Nicholas J. Foster
SENCR-MIC Laboratory Technician
Fayetteville State University
Office: (910) 672-2039
Probe Lab: (910) 672-2037
Email: nfoster1-at-uncfsu.edu

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4, 33 -- Date: Tue, 27 May 2014 14:28:35 -0400
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Nick,

We get ours from Buehler, Part Number: 40-7222, 10 pack of 12" Microcloth with adhesive on the back.

--John

John Chandler
EM Lab
jpchandl-at-mines.edu
303.384.2203

On May 27, 2014, at 12:33 PM, {nfoster1-at-uncfsu.edu} {nfoster1-at-uncfsu.edu} wrote:

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} Via Allen Glazner at UNC
}
} I have resurrected an ancient Buehler Vibromet polisher (model 67-1517) and would like to use it to
} put a final polish on probe samples. I have been unable to find cloths or abrasive pads for it (12-inch model)
} and wonder if anyone has experience with this polisher.
}
} Nick
}
} Nicholas J. Foster
} SENCR-MIC Laboratory Technician
} Fayetteville State University
} Office: (910) 672-2039
} Probe Lab: (910) 672-2037
} Email: nfoster1-at-uncfsu.edu
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} 4, 33 -- To: "Microscopy-at-microscopy.com" {Microscopy-at-microscopy.com}
} 4, 33 -- Date: Tue, 27 May 2014 14:28:35 -0400
} 4, 33 -- Subject: Buehler Vibromet polisher help.
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10, 27 -- From jpchandl-at-mines.edu Tue May 27 13:37:47 2014
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10, 27 -- From: John Chandler {jpchandl-at-mines.edu}
10, 27 -- To: "nfoster1-at-uncfsu.edu" {nfoster1-at-uncfsu.edu}
10, 27 -- CC: "microscopy-at-microscopy.com" {microscopy-at-microscopy.com}
10, 27 -- Subject: Re: [Microscopy] Buehler Vibromet polisher help.
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Are you sure you want to do that? What kind of samples do you process?

My experience is that vibratory polishing with colloidal silica can lead to topography that causes problems rather than resolving problems. Differences in hardness can lead to polishing relief. It can also chemically etch some samples. And there can also be a problem of silica residue on the sample after polishing if the cleaning protocol is not thorough enough. It is quite difficult to remove those deposits once they form.

Warren Straszheim
________________________________________
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To: Straszheim, Warren E [BIOTC]

Via Allen Glazner at UNC

I have resurrected an ancient Buehler Vibromet polisher (model 67-1517) and would like to use it to
put a final polish on probe samples. I have been unable to find cloths or abrasive pads for it (12-inch model)
and wonder if anyone has experience with this polisher.

Nick

Nicholas J. Foster
SENCR-MIC Laboratory Technician
Fayetteville State University
Office: (910) 672-2039
Probe Lab: (910) 672-2037
Email: nfoster1-at-uncfsu.edu

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Title-Subject: [Filtered] Rotators for Cold Rooms

Message: Hello to everyone in Microscopy Land. Our old rotator has finally died and we can not get
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Title-Subject: [Filtered] Reminder : workshop registration closing

Message: UMB Current Electron Microscopy Techniques Workshop

June 2-3, 2014, University of Maryland, Baltimore

Dear Colleagues,

This serves as a reminder that registration for the UMB Current EM Techniques Workshop will close
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Historically, I have manually performed the typical two element energy calibration. Afterwards, I can collect objective peak location information which I have logged as evidence of the calibration. I evaluate and record the "as found" condition, perform the energy calibration (if needed), then evaluate and record "as left" information.

Now we have a new system that is totally automated and requires only one element for calibration. It works extremely well as I judge by eye, but I also need unbiased, objective data to document that the calibration "worked", which I cannot access from the system.

We are considering using a certified two element solid solution material as a control sample, but I would like to know what other methods may be typical within the industry. What is everyone else doing for quality assurance with respect to energy calibration?

Thanks,

Chris Holp
FENOC
BETA Labs

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Hi Chris

In the days before we had ultra thin windows the two element calibration
method, usually using aluminium and copper, was almost universal. Now we
have thinner windows and we need to have a low energy peak within the new
area that has been opened up to us, so using a copper K and L line has now
become the norm.

Most of us just accept the data as corrected by the system, but we are able
to check for accuracy against other very low energy peaks in our spectrum.
That is the simple route now what do others say?

Regards

Steve

Steve Chapman FRMS
Protrain for Consultancy and Courses in Electron Microscopy
Mobile +44 (0) 7711 606967 Fax +44 (0)1280 814007
www.emcourses.com

-----Original Message-----
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To: protrain-at-emcourses.com

Nick,

When I worked at NASA GRC we used the vibratory polishers for final
polishing of microprobe samples. Instead of collodial silica however we
used something we called 'Magic Dust'. It is 0.5 micron diamond powder
available from Kay Diamond Products. Unlike collodial silica it would
keep the samples flat with little contamination. After installing a
cloth we would use a diamond extender lubricant from Metlab Corp then
sprinkle the Magic Dust on the cloth and put the samples on using the
weighted holders for 1 to 2 hours.

No financial interest in Kay or Metlab. Just happy customers for over
20 years.

nfoster1-at-uncfsu.edu wrote:
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} I have resurrected an ancient Buehler Vibromet polisher (model 67-1517) and would like to use it to
} put a final polish on probe samples. I have been unable to find cloths or abrasive pads for it (12-inch model)
} and wonder if anyone has experience with this polisher.
}
} Nick
}
} Nicholas J. Foster
} SENCR-MIC Laboratory Technician
} Fayetteville State University
} Office: (910) 672-2039
} Probe Lab: (910) 672-2037
} Email: nfoster1-at-uncfsu.edu
}
} ==============================Original Headers==============================
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Inter/Micro 2014
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http://www.mcri.org/home/section/101-915/inter-micro-2014

Greetings!
Please join us for our 66th annual microscopy conference held at McCrone
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Location
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.........................................
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Title-Subject: [Filtered] monitor for a JEOL 8900 microprobe

Message: the left monitor on our JXA 8900 microprobe has given up the ghost. Since we are self
insured, and really handy, we were wondering if anyone out there has a doner JEOL microprobe or SEM
with a monitor that needs a new and loving home. we will pick up the cost of shipping and if you
want compensation we can negotiate. If you are in the north east we can even make a road trip, it is
pretty desolate up here.

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We are in the process of switching for the first time from a tungsten to a LaB6 emitter on our Tecnai Biotwin 12, circa 2006. I have gotten varying opinions about whether we should shut off the emitter every time we change samples. On our Biotwin (perhaps on all Tecnai 12's?) the gun does not seem to be isolated from the column during specimen exchange (V5 stays open to maintain pumping by the single ion pump), so we are concerned that blowing vacuum during sample exchange (as happens from time to time) could destroy our expensive filament. On the other hand, I have been told that it's not good to turn the LaB6 on and off frequently, as would be necessary to load multiple samples during a session.

I am interested in hearing your experience running a LaB6 on this instrument: do you shut off the beam each time you change samples, and has the emitter been damaged when you have blown vacuum on the column? Also, how long have your emitters been lasting? Thanks!

Marie

Dr. Marie E. Cantino
Associate Professor of Physiology and Neurobiology
Director, Electron Microscopy Laboratory
University of Connecticut, Unit 3242
Phone: 860-486-3588
Fax: 860-486-6369

==============================Original Headers==============================
6, 17 -- From marie.cantino-at-uconn.edu Thu May 29 09:05:29 2014
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6, 17 -- Subject: LaB6 on Tecnai Biotwin
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Dear Dr. Cantino,
We have been running a LaB6 emitter in our Philips CM12 for over
20 years. When changing samples we leave the beam saturated. Even the
most experienced users and well-trained newer users will, on infrequent
occasions, encounter a situation when enough air enters the column
after prepumping to cause the high tension to go off and on rare
occasions to cause vacuum valves to close as part of the fail/safe
system. We believe the impact on the life of the LaB6 emitter is
minimal compared to what daily multiple saturation and desaturation
would cause.

Typically, our emitters last more than 2000 hours of saturated
beam time. Incidentally, it was recommended to us by our service
engineer that we leave the high tension on and the beam desaturated at
all times when not in use.

Hope this helps, Don

Donald Gantz, MS
Dept. Physiology & Biophysics
Boston Univ. School of Medicine
700 Albany Street
Boston, MA 02118
617-638-4017

On 5/29/2014 10:06 AM, marie.cantino-at-uconn.edu wrote:
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} We are in the process of switching for the first time from a tungsten to a LaB6 emitter on our Tecnai Biotwin 12, circa 2006. I have gotten varying opinions about whether we should shut off the emitter every time we change samples. On our Biotwin (perhaps on all Tecnai 12's?) the gun does not seem to be isolated from the column during specimen exchange (V5 stays open to maintain pumping by the single ion pump), so we are concerned that blowing vacuum during sample exchange (as happens from time to time) could destroy our expensive filament. On the other hand, I have been told that it's not good to turn the LaB6 on and off frequently, as would be necessary to load multiple samples during a session.
}
} I am interested in hearing your experience running a LaB6 on this instrument: do you shut off the beam each time you change samples, and has the emitter been damaged when you have blown vacuum on the column? Also, how long have your emitters been lasting? Thanks!
}
} Marie
}
} Dr. Marie E. Cantino
} Associate Professor of Physiology and Neurobiology
} Director, Electron Microscopy Laboratory
} University of Connecticut, Unit 3242
} Phone: 860-486-3588
} Fax: 860-486-6369
}
}
}
} ==============================Original Headers==============================
} 6, 17 -- From marie.cantino-at-uconn.edu Thu May 29 09:05:29 2014
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} 6, 17 -- From: Marie Cantino {marie.cantino-at-uconn.edu}
} 6, 17 -- Content-Type: text/plain; charset=us-ascii
} 6, 17 -- Subject: LaB6 on Tecnai Biotwin
} 6, 17 -- Date: Thu, 29 May 2014 10:05:28 -0400
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6, 24 -- From gantz-at-bu.edu Thu May 29 10:38:40 2014
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Dear Marie,

We run one or two instrument on LaB6 emitters - a Philips CM30 (300kV)
and a JEOL 200CX (200kV).

We always turn the heating current and high tension off for every sample
exchange for two reasons. First, there is a substantial danger that you
will get a beam of X-rays emerging from the goniometer, especially if
the beam is on and the HT is active. We have measured X-ray flux of
several hundred micro-Sieverts per hour coming from X-rays emerging from
the (empty) goniometer with the beam on, several micro-amps of beam
current and the objective aperture left in. With older instruments this
is a significant problem (and enough to warrant inspection from the
Radiation Safety Officer). If the HT and beam is off, there is no risk
at all. Yes, it is a mild inconvenience, but it beats the risk of
inadvertently radiating our users.

Second, LaB6 filaments tend to oxidise over time so having a puff of air
go up into the gun with a hot filament is unwise. We have about 30-40
users on the instrument and everyone has, at one time, vented the vacuum
system during a sample extract. It is a matter of when not if.

The last LaB6 filament we had lasted 3600 hours or about over a year and
a half of use.

Yours, Jon

Department of Materials Science & Metallurgy, University of Cambridge,
United Kingdom

On 2014-05-29 15:11, marie.cantino-at-uconn.edu wrote:
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} We are in the process of switching for the first time from a tungsten
} to a LaB6 emitter on our Tecnai Biotwin 12, circa 2006. I have gotten
} varying opinions about whether we should shut off the emitter every
} time we change samples. On our Biotwin (perhaps on all Tecnai 12's?)
} the gun does not seem to be isolated from the column during specimen
} exchange (V5 stays open to maintain pumping by the single ion pump),
} so we are concerned that blowing vacuum during sample exchange (as
} happens from time to time) could destroy our expensive filament. On
} the other hand, I have been told that it's not good to turn the LaB6
} on and off frequently, as would be necessary to load multiple samples
} during a session.
}
} I am interested in hearing your experience running a LaB6 on this
} instrument: do you shut off the beam each time you change samples, and
} has the emitter been damaged when you have blown vacuum on the column?
} Also, how long have your emitters been lasting? Thanks!
}
} Marie
}
} Dr. Marie E. Cantino
} Associate Professor of Physiology and Neurobiology
} Director, Electron Microscopy Laboratory
} University of Connecticut, Unit 3242
} Phone: 860-486-3588
} Fax: 860-486-6369
}
}
}
} ==============================Original
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From meds-canadian2-at-nationalcablenetworks.ru Thu May 29 11:35:08 2014
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This topic has been discussed a few times, I guess, but I am happy to forward our (overall very positive) experience with LaB6 on FEI 120kV TEM's.
Our CM12 is operated with a LaB6 of one manufacturer, at 120kV, always, exclusively, without any problems. Contrast -at-120kV is fine and depends very much on the sample preparation and the specimen quality and ... (also on the voltage, I know, but ...), and also on the quality of the CCD camera and its settings ...
Since } 20yrs, we decrease the Filament (= emission) from 18 to 20 down to Zero, during sample exchange, and raising back to 18 to 20, after inserting the holder (and IGP below around 20 or so); that's all. High voltage is left on at 120kV, always, during day time, even when inserting a cryo holder. Really no problems at all, here, on a TEM CM12 used by } 20 users over the years. Our LaB6 filaments (manufacturer on request only) last for } 3 years, the present one for almost four years, with 5 days-a-week-use, 8-12 hours per day.
Vacuum on our TEM is really good, with IGP at 5 to 10 (liquid nitrogen is used every day, every hour). We do not open the column ... as we record images fully digitally (no films, since 1998), and the LaB6 stays in the column for 3 to 4 years. This is advantageous ...
only positive experience with LaB6 on a TEM of this kind, for bio-samples of all kinds.
It also depends on the training of the users, what 'high vacuum' really means, how to achieve it, proper starting of the TEM in the morning, cryo-cycle every night (= IGP off for 4 hours), and so on.
kind regards,
Reinhard

Prof. Dr. Reinhard Rachel
University of Regensburg
Centre for EM / Anatomy
Faculty of Biology & Preclin. Med.
Universitaetsstrasse 31
D-93053 Regensburg - Germany
tel +49 941 943 2837, 1720
fax +49 941 943 2868
mail reinhard.rachel-at-biologie.uni-regensburg.de
office: VKL 3.1.29

Next microscopy conference:
http://www.imc2014.com/
18th IMC 2014 in Prague, Czech Rep.

==============================Original Headers==============================
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From pharmacy_popular6-at-mgts.by Fri May 30 07:04:47 2014
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Hello,

I would like to take STEM-EDX maps acquired using Bruker Esprit and open
the raw spectral data in Matlab for more hands-on processing of the
spectrum images. Has
anyone had experience converting the BCF file to another format or
importing directly into Matlab? Ideally I would like to convert the product
back to BCF as well to make use of what the Esprit software does well.

Thanks!
Andrew

==============================Original Headers==============================
3, 35 -- From awagner-at-umn.edu Fri May 30 08:00:39 2014
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3, 35 -- Subject: Opening Bruker Esprit EDX HyperMaps in Matlab?
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Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

I work with an Oxford Aztec that also takes spectral maps. I've also had users ask about taking the data in a portable format that they can work with it on their own. I generally discourage them. At a minimum, I try to discuss with them at length what they have in mind for the data. Many times, it seems that they don't know what they want to do with the data, but they'd like to have it anyway.

Most do not seem to appreciate what would be involved in processing the data. In fact, I had one Chinese student from Civil Engineering ask yesterday if he could get text versions of all his spectra. There are only about 30 spectra, so that isn't too nasty a request. I took one spectrum, exported it in MSA format, imported into Excel, and made a chart. He saw the shape as before but then asked which peaks were which. I showed him where in the MSA file he could find the peak labels and locations including the 4 different peaks for Ca. Then the question came up about how to quantify the data into weight fractions. I did not cover background modeling and removal, nor did I discuss peak deconvolution or integration. I did explain that he would have to have a standard for each of the elements so he could compare his sample intensity to the standard intensity. I did not launch into the necessary matrix corrections. I did tell him that his sample violated many of the assumptions necessary for quant analysis. What I did tell him was enough to dissuade him for following through on his request.

I would go through the same exercise with spectral images. The vendors do a lot in their software and I am hesitant to try to reproduce it. Maybe I am too old and lacking creative juices, but I find it hard to believe that I could seriously improve on what is available. I could see exporting the map images and doing some math on them to try to tease out phases. That would be a much simpler data set to work with.

If you are still set on doing your own processing, I recall NIST was promoting a format for spectral images. They also had developed some routines for basic processes. I have not followed it in recent years, so I cannot tell you where things stand. Maybe someone else can chime in.

Warren Straszheim

-----Original Message-----
X-from: awagner-at-umn.edu [mailto:awagner-at-umn.edu]
Sent: Friday, May 30, 2014 8:01 AM
To: Straszheim, Warren E [BIOTC]

Hello,

I would like to take STEM-EDX maps acquired using Bruker Esprit and open the raw spectral data in Matlab for more hands-on processing of the spectrum images. Has anyone had experience converting the BCF file to another format or importing directly into Matlab? Ideally I would like to convert the product back to BCF as well to make use of what the Esprit software does well.

Thanks!
Andrew

==============================Original Headers==============================
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3, 35 -- Subject: Opening Bruker Esprit EDX HyperMaps in Matlab?
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==============================Original Headers==============================
14, 36 -- From wesaia-at-iastate.edu Fri May 30 10:04:56 2014
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Dear List Members,

We are organizing the scientific event:
"Materials in Science and Technology 2014 (MIST 2014)".
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MIST 2014 includes two international conferences and one autumn school:
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Specifically, the Autumn School, AISMAT 2014 (http://aismat.org), after a
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Hi Andrew,

Bruker have a built in facility to output the data in portable form. In
Esprit you can save out data as a "Raw database file" from the File type
dropdown menu. This will then save the dataset into a raw binary
datacube x,y,energy, that can be imported into Matlab/IDL/whatever...
You will need to be familiar with how to import raw binary files. I've
done this for Bruker files in the past but of the top of my head I can't
remember the order (x,y or energy first?) or the bit depth used, but a
bit of playing around should find this out for you (as you already know
the data dimensions).

A word of warning however, Bruker save there data in countwise, i.e. for
each X-ray counts they place 'energy,x,y' and entry into their datafile.
This results in very memory efficient storage. A datacube (such as the
"Raw database file") has simply the dimensions of the datacube and as
such can be very space inefficient, especially for sparsely sampled
data. For example one of my 6Mb Bruker files expanded to 1GB in raw form!

I can't help you with loading the data back in to Esprit after that
however (as you cannot import Raw Database files back into the software)
this 'one-way' nature is not uncommon as manufacturers formats contains
much information, and simulating/re-generating it would be very difficult!

Matthew

On 30/05/2014 11:12 PM, awagner-at-umn.edu wrote:
} Hello,
}
} I would like to take STEM-EDX maps acquired using Bruker Esprit and open
} the raw spectral data in Matlab for more hands-on processing of the
} spectrum images. Has
} anyone had experience converting the BCF file to another format or
} importing directly into Matlab? Ideally I would like to convert the product
} back to BCF as well to make use of what the Esprit software does well.
}
} Thanks!
} Andrew
}

--
Dr M.Weyland, Associate Professor & Titan Manager
--------------------------------------------------------------------------
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Victoria 3800
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--------------------------------------------------------------------------
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Thanks Matthew!

I actually figured it out yesterday with some helpful comments from
another listserv member and it's just as you said. The format is
Lispix ( http://www.nist.gov/lispix/ and for those who use EDX but
might not be aware, see also HyperSpy, http://hyperspy.org/ and
DTSA-II, http://www.cstl.nist.gov/div837/837.02/epq/dtsa2/ ). It is a
raw binary format with each bit of data in a continuous stream, i.e.
no delimiter. Each channel of the first pixel is followed by each
channel of the second pixel (left to right, top to bottom). An
associated "rpl" file tells you the channel depth per pixel and the
height and width of the spectrum image. It is uncalibrated (in space
and energy) so that must be taken care of by hand. Bruker opts for
8-bit data, so if you happen to have more than 255 counts in any
channel you will have trouble. Esprit appears to truncate the data at
the last channel with any counts for sake of file size, so the channel
depth is important and potentially variable.

Lispix works great for spectra analysis and moving this data into
DTSA. I have not played much with HyperSpy yet. On the off chance
anyone else is interested in getting the data into Matlab, feel free
to contact me. I put together a matlab script to read the file into a
datacube and also perform some basic processing. Simple, but if it
saves someone some time in Matlab, great. I am looking at some
exceptionally beam sensitive specimens, so I am playing with probe
size, step size, dwell time, and probe current to see what compromises
I can make while being able to spatially resolve changes in relative
composition. That's where Matlab comes in as I'd like to do running
sums of the spectra while also doing 2-D running pixel sums as opposed
to just binning or increasing probe size or step size.

Some nice discussion of EDX sensitivity, SDD detectors, and approaches
to analysis can be found in this powerpoint:
http://epmalab.uoregon.edu/Workshop2/DonovanWorkshop07_SDD_Newbury.pdf
and this associated paper:
http://www.geology.wisc.edu/~johnf/g777/Scanning/NewburyBright-2005.pdf

Cheers,
Andrew

On Sun, Jun 1, 2014 at 5:52 PM, Matthew Weyland
{matthew.weyland-at-monash.edu} wrote:
} Hi Andrew,
}
} Bruker have a built in facility to output the data in portable form. In
} Esprit you can save out data as a "Raw database file" from the File type
} dropdown menu. This will then save the dataset into a raw binary datacube
} x,y,energy, that can be imported into Matlab/IDL/whatever... You will need
} to be familiar with how to import raw binary files. I've done this for
} Bruker files in the past but of the top of my head I can't remember the
} order (x,y or energy first?) or the bit depth used, but a bit of playing
} around should find this out for you (as you already know the data
} dimensions).
}
} A word of warning however, Bruker save there data in countwise, i.e. for
} each X-ray counts they place 'energy,x,y' and entry into their datafile.
} This results in very memory efficient storage. A datacube (such as the "Raw
} database file") has simply the dimensions of the datacube and as such can be
} very space inefficient, especially for sparsely sampled data. For example
} one of my 6Mb Bruker files expanded to 1GB in raw form!
}
} I can't help you with loading the data back in to Esprit after that however
} (as you cannot import Raw Database files back into the software) this
} 'one-way' nature is not uncommon as manufacturers formats contains much
} information, and simulating/re-generating it would be very difficult!
}
} Matthew
}
}
} On 30/05/2014 11:12 PM, awagner-at-umn.edu wrote:
} }
} } Hello,
} }
} } I would like to take STEM-EDX maps acquired using Bruker Esprit and open
} } the raw spectral data in Matlab for more hands-on processing of the
} } spectrum images. Has
} } anyone had experience converting the BCF file to another format or
} } importing directly into Matlab? Ideally I would like to convert the
} } product
} } back to BCF as well to make use of what the Esprit software does well.
} }
} } Thanks!
} } Andrew
} }
}
} --
} Dr M.Weyland, Associate Professor & Titan Manager
} --------------------------------------------------------------------------
} Monash Centre for Electron Microscopy
} Bldg 81
} Monash University
} Victoria 3800
} Australia.
} --------------------------------------------------------------------------
} www.mcem.monash.edu.au
} -------------------------------------------------------------------------
} Ph: (+61) 3 990 59026 --- Fax: (+61) 3 990 53600
} --------------------------------------------------------------------------
}

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realname - Dr. Connie Cummings
Email - ultrapathimaging-at-gmail.com
ORGANIZATION - DBA UltraPath Imaging
EDUCATION - Graduate College
LOCATION - Durham, NC USA
SUBJECT_OF_QUESTION - EM Speakers/EM lab coaches
QUESTION - Good evening all,
I am the ultrastructural pathologist who contracts my services to the
EM laboratory at NIEHS/NTP, the campus in Research Park, Durham, NC.
My question is: We are looking for anyone with EM experience that would
be willing to come to NC and spend 2 to 5 days in our EM Lab. Our
objective for this visit would be to inspect the lab followed by a list
of pertinent items that could be changed. If there are MSA members out
there, I would certainly like to hear from you. I hope everyone is
having a nice evening.
Connie

my phone number 919-316-499
Another contacts:
Dr. Herbert's #919-5414613

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Hi Andrew,

Last year, I made a video and an ImageJ plugin for doing this for stacks acquired from the Bruker EDS on the Titan TEM at NCEM. Â

So here is about a 15 minute video where I import the data into ImageJ, make maps, make an RGB, export a spectrum from ImageJ and load it in a spectrum processing software:

https://berkeley.box.com/s/bya3oaiqat25hxwrsate

And here are the links to the source code for the ImageJ plugin which you can use as a starting point for whatever you want to do:

https://berkeley.box.com/s/bd74bnmg405lppfmfakd

and

https://berkeley.box.com/s/brtal2oz4hztj8wfws3y

I hope you find it helpful,

Zack Gainsforth
Space Sciences Laboratory, UC Berkeley
7 Gauss Way
Berkeley, CA Â 94720
cell: 626-437-9186
zackg-at-ssl.berkeley.edu

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Email: vakimler-at-med.wayne.edu
Name: Vickie Kimler

Organization: Wayne State University School of Medicine

Title-Subject: [Filtered] Optimum Embedding Medium for Tissue Cells with
Lipid Droplets

Message: Does anyone have an opinion on the optimum embedding medium and
its respective physical characteristics (hardness, pH, etc.) for TEM on
tissue cells (explants) which contain lipid droplets? We have been using
Embed-812 and Durcupan interchangeably. Sometimes our resolution on
small droplets is poor and we want to enhance this.

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Hi all

As I'm trying to make reworking some "old" optical microscope, I come
with a few questions to solve one problem.

I've a set of Zeiss Epiplan OL from the 70" (designed for the Standard
et Universal generation), but coming from the metallographic microscope
mounted on a Siemens Microprobe. They were factory modified to be
mounted in vacuum on a special turret while the microscope body was
fixed on the column on the air side. The body of the objectives is
shorter than the standard one. As I want to use them on a
Photomicroscope, I need to machine extension tubes with 2 RMS threads.

My questions are about the parfocal length of these OL.
-First, does someone know if these objectives did exist in 33 and
45 mm parfocal length, or only in 45 mm ?
-Secondly, how could I determine this parfocal length, which I need
to know the length for the extension tubes.

As I know a working distance for each OL (from a Zeiss doc), I can
calculate two possible extension lengths. I tried some tests on a
optical bench, fixing a 160 mm tube length and trying to determine the
length between the thread plane of that virtual tube and the back of the
lens, but as the image depth become very long with high power obectives,
it is very difficult to locate the best focus position.
Could it be possible to see something reliable looking at distortion and
chromatic aberation near the border of the field ?

All other idees are wellcome !
Thanks

--

J. Faerber
IPCMS-DSI
Institut de Physique et Chimie des Matériaux de Strasbourg
Département Surfaces et Interfaces
23, rue de Loess ; BP43
67034 Strasbourg CEDEX 2
France

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

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Title-Subject: [Filtered] operating manual

Message: Hi

My friend is looking for operating manuals for FEI TEM T-20 and HRTEM F-30

Best wishes
Rashmi

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Title-Subject: [Filtered] Spares for EM

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We are looking for the procedure to buy the spares directly from FEI for
ESEM, TEM and HRTEM

best wishes
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Hi John

Your are fully right, John !

The problem is that these objectives have a modified body which is much
shorter then normaly. That's why I don't know for which parfocal
distance they were originaly build. The extention tubes I want to
machine will in fact replace the missing body.
Following the Universal/phomi documentation from Zeiss, it seems that
both 33 and 45 mm parfocal distances epicondenser/objectives have been
provided.
I don't know if my explanation is clear ! A picture would be simpler.

Regards

jacques

J. Faerber
IPCMS-DSI
Institut de Physique et Chimie des MatÃ©riaux de Strasbourg
DÃ©partement Surfaces et Interfaces
23, rue de Loess ; BP43
67034 Strasbourg CEDEX 2
France

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

Le 03/06/2014 15:51, John Mitchels a Ã©crit :
} Hi Jacques
}
} I am a little confused. The parfocal length of an objective is simply
} the length from the base of the thread seat to end of the objective
} plus the working distance. Provided this distance is maintained all
} you need do is adjust the mouning rings.
}
} Anythoughts
} John
}
} On 3 June 2014 10:29, {jacques.faerber-at-ipcms.u-strasbg.fr} wrote:
} }
} }
} } ----------------------------------------------------------------------------
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} } Hi all
} }
} } As I'm trying to make reworking some "old" optical microscope, I come
} } with a few questions to solve one problem.
} }
} } I've a set of Zeiss Epiplan OL from the 70" (designed for the Standard
} } et Universal generation), but coming from the metallographic microscope
} } mounted on a Siemens Microprobe. They were factory modified to be
} } mounted in vacuum on a special turret while the microscope body was
} } fixed on the column on the air side. The body of the objectives is
} } shorter than the standard one. As I want to use them on a
} } Photomicroscope, I need to machine extension tubes with 2 RMS threads.
} }
} } My questions are about the parfocal length of these OL.
} } -First, does someone know if these objectives did exist in 33 and
} } 45 mm parfocal length, or only in 45 mm ?
} } -Secondly, how could I determine this parfocal length, which I need
} } to know the length for the extension tubes.
} }
} } As I know a working distance for each OL (from a Zeiss doc), I can
} } calculate two possible extension lengths. I tried some tests on a
} } optical bench, fixing a 160 mm tube length and trying to determine the
} } length between the thread plane of that virtual tube and the back of the
} } lens, but as the image depth become very long with high power obectives,
} } it is very difficult to locate the best focus position.
} } Could it be possible to see something reliable looking at distortion and
} } chromatic aberation near the border of the field ?
} }
} } All other idees are wellcome !
} } Thanks
} }
} } --
} }
} } J. Faerber
} } IPCMS-DSI
} } Institut de Physique et Chimie des MatÃ©riaux de Strasbourg
} } DÃ©partement Surfaces et Interfaces
} } 23, rue de Loess ; BP43
} } 67034 Strasbourg CEDEX 2
} } France
} }
} } Tel 00 33(0)3 88 10 71 01
} } Fax 00 33(0)3 88 10 72 48
} } E-mail : Jacques.Faerber-at-ipcms.unistra.fr
} }
} }
} } ==============================Original Headers==============================
} } 10, 24 -- From jacques.faerber-at-ipcms.u-strasbg.fr Tue Jun 3 04:23:49 2014
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9, 26 -- From jacques.faerber-at-ipcms.u-strasbg.fr Tue Jun 3 10:15:03 2014
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9, 26 -- OL
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Title-Subject: [Filtered] SEM for Cornea

Message: Some one is asking me to do SEM on his samples of cornea. I do
SEM on metals, semiconductors and so on normally. I do not have much
experiences on this kind of biological samples.

If some one on this list server knows how to do it, please tell me.

And I have a SEM of Fei XL-30 available for this sample.

Thanks in advance.

Zhenquan Liu

Arizona State University

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Dear Zhenquan,

the preparation of biological material for SEM depends very much of the instruments you have at your disposal.
Unlike metals and semiconductors, biological material contains a lot of water and, because you have to remove it for SEM, the material loses its structure and tends to be very unstable under high vacuum. For this reason the biologists need critical point dryers for example. Hopefully you have access to such instruments as well as to the person with the expertise to use it.

By googling with "preparation cornea for EM", I found the first link to be interesting:
http://www.optics.rochester.edu/workgroups/cml/opt307/spr05/anant/

best regards,
Stephane

--------------------------------------------
On Wed, 6/4/14, zaluzec-at-microscopy.com {zaluzec-at-microscopy.com} wrote:

Subject: [Microscopy] viaWWW:SEM for Cornea
To: nizets2-at-yahoo.com
Date: Wednesday, June 4, 2014, 1:00 PM

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Title-Subject: [Filtered] SEM for Cornea

Message: Some one is asking me to do SEM on his samples of
cornea. I do
SEM on metals, semiconductors and so on normally. I do not
have much
experiences on this kind of biological samples.

If some one on this list server knows how to do it, please
tell me.

And I have a SEM of Fei XL-30 available for this sample.

Thanks in advance.

Zhenquan Liu

Arizona State University

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

Dear Listers,

I have a user on FIB620 FIB/SEM with old pre-Xp dual-beam control
software (Ver. 6.3 from FEI/Phillips), who needs to find out how many
dwell points would be made by ion beam on 1um long line.

The calculation would be straight forward if we knew overlap (available
in material file or on the screen) and the beam diameter assumed by the
software, but... the assumed beam diameter is hidden somewhere.

Obviously such information can be derived by application of good old
hacking and some investment of time, but I was wondering if there is
anyone out there who already knows where does software store beam
diameter parameters.

Thank you very much beforehand,

--
Valery Ray
=================================
PBS&T, MEO Engineering Co., Inc.
290 Broadway, Suite 298
Methuen, MA 01844 USA
Phone: +1-978-296-5063 - leave a message with call-back number
US Mobile: +1-978-305-0479
Skype: pbstmeo
E-mail: vray-at-partbeamsystech.com
Web: www.partbeamsystech.com
Web: www.freudlabs.com

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

I am trying to correlate super resolution imaging with TEM on Tokuyasu frozen sections. As an internal marker in fluorescence, we would like to stain the the nuclei before cryoprotection/freezing. I would appreciate any recommendations for a rather robust vital nuclear stain. The STED imaging is done in green channel.

Thanks, Michal
--

Michal Jarnik, Ph.D.

Electron Microscope Facility

Cell Biology and Metabolism Program

National Institute of Child Health and Human Development National Institutes of Health

9000 Rockville Pike, Bldg. 18T, Room 101 Bethesda, MD 20892

==============================Original Headers==============================
9, 30 -- From michal.jarnik-at-nih.gov Wed Jun 4 11:30:04 2014
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9, 30 -- From: "Jarnik, Michal (NIH/NICHD) [E]" {michal.jarnik-at-nih.gov}
9, 30 -- To: "Microscopy-at-Microscopy.com" {Microscopy-at-microscopy.com}
9, 30 -- Subject: Vital DNA stain
9, 30 -- Thread-Topic: Vital DNA stain
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From cheapest_drugs9-at-rr.com Wed Jun 4 11:36:41 2014
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Have you tried DRAQ5? You could excite the dye with a 488 nm laser - Is that what you mean "green channel"?

Zhaojie Zhang
University of Wyoming

-----Original Message-----
X-from: michal.jarnik-at-nih.gov [mailto:michal.jarnik-at-nih.gov]
Sent: Wednesday, June 04, 2014 10:33 AM
To: Z.J. Zhang

I am trying to correlate super resolution imaging with TEM on Tokuyasu frozen sections. As an internal marker in fluorescence, we would like to stain the the nuclei before cryoprotection/freezing. I would appreciate any recommendations for a rather robust vital nuclear stain. The STED imaging is done in green channel.

Thanks, Michal
--

Michal Jarnik, Ph.D.

Electron Microscope Facility

Cell Biology and Metabolism Program

National Institute of Child Health and Human Development National Institutes of Health

9000 Rockville Pike, Bldg. 18T, Room 101 Bethesda, MD 20892

==============================Original Headers==============================
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18, 36 -- From ZZhang-at-uwyo.edu Wed Jun 4 11:46:52 2014
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DRAQ5 is far red, so 488 excitation would be far from optimal. What about SYBR Green?

Jill Pflugheber
St. Lawrence Universtiy

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Sent: Wednesday, June 04, 2014 12:54 PM
To: Jill Pflugheber

Have you tried DRAQ5? You could excite the dye with a 488 nm laser - Is that what you mean "green channel"?

Zhaojie Zhang
University of Wyoming

-----Original Message-----
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Sent: Wednesday, June 04, 2014 10:33 AM
To: Z.J. Zhang

I am trying to correlate super resolution imaging with TEM on Tokuyasu frozen sections. As an internal marker in fluorescence, we would like to stain the the nuclei before cryoprotection/freezing. I would appreciate any recommendations for a rather robust vital nuclear stain. The STED imaging is done in green channel.

Thanks, Michal
--

Michal Jarnik, Ph.D.

Electron Microscope Facility

Cell Biology and Metabolism Program

National Institute of Child Health and Human Development National Institutes of Health

9000 Rockville Pike, Bldg. 18T, Room 101 Bethesda, MD 20892

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Hello listers.

I have scanned in all the old images that I thought were good and now want to get rid of the TEM and SEM negatives I have had for many years. Does anyone know if they are recyclable?

Thanks
JoAnn Buchanan
Stanford University School of Medicine

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Hi JoAnn.

Yes, your TEM and SEM negatives are recyclable. Rochester Silver Works (http://www.rochestersilverworks.com/) does this. For many years they were part of Eastman Kodak and they recycled over 50 years of negatives from our lab. I suspect that there are other recyclers who do as well.

Note: I have no financial interest in Rochester Silver Works. I simply used them to be environmentally responsible.

Best regards,
John Minter
Kodak Analytical Sciences

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Email: sbras-at-uw.edu
Name: Scott Braswell

Organization: University of Washington

Title-Subject: [Filtered] In-situ NP measurement

Message: Hi,

I received an email yesterday asking me to characterize the size of FeOx
nanoparticles in strong NaHO. I wonder if drying the solution would
result in too much salt. My thoughts are to either try cryoTEM or use a
liquid-cell TEM holder. Any insight you can offer for this sample prep
would be appreciated.

Thanks,
Scott

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Email: zhenquan.liu-at-asu.edu
Name: Zhenquan Liu

Organization: Arizona State University

Title-Subject: [Filtered] Wet method on Evironmental SEM for cornea

Message: Hi, Everyone,

Thanks a lot for replying from several people for my previous email
which asking for a way to do SEM on cornea. I have learned a lot form
these replies. There are so many things I have to go through sample
preparation and actually we do not have most of the instruments in our
lab, but there are available in School of Biology.

I read these replies carefully and I did not see anyone who mentioned
wet method on environmental SEM to get images from cornea. We have Fei
XL-30 Environmental SEM. We can do wet method.

A wet method is to introduce some water vapor near the sample to keep
sample wet. The humidity can be controlled to 100%.

I think that it might be difficult to do so. On this list server there
are many experts in biology field, but none of them mentioned wet
method. I think if it is a good way to work on cornea, people would
have told me. Many years ago I tried wet method on the surface of the
samples in water. I did not get much useful information. I might miss
some thing when I did it.

Therefore if someone has some experiences on wet method, even better on
cornea, please tell me. It will be great.

Thanks to Stephane, Lee Cohen-Gould, Debra Sherman and Ed Haller and
more people who replied to my previous email.

Zhenquan Liu
Arizona State University

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We have an FEI Quanta and could probably work in the same regime as you can. Also like you, I am a materials scientist, not a biologist, but I get the occasional question to look at biological material. Most recently, it was an aggregate of bacteria. The request came from a Civil/Environmental Engineer so I don't think they understood what they were asking nor would they quite know how to interpret what I found, if I had found something. That should be the first question - What are they looking for and what do they expect it to look like?

A lot of biological material will not be that interesting given all the water present and the organic membranes. In my little experience, the unprepared samples just don't have much contrast when you are looking at the outside. It is cell wall and maybe a film of water. I would be interested in hearing what the biologists have to say about imparting contrast. Of course, the structure should be much more interesting on the inside but I suppose it needs enhancement, even then.

I understand it is technically possible to keep a sample from dehydrating. The triple point of water is at 0.006 atm or 600 Pa at near 0C, so it should be possible to maintain a saturated atmosphere. Of course, that requires a cooling stage. If you are working at room temperature, that number rises to about 2300 Pa. That presents more of a challenge. I was not able to keep my bacteria granule from drying out in the SEM.

I'll be interested in hearing any recipes that prove helpful.

Warren Straszheim
Iowa State University

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Sent: Wednesday, June 04, 2014 8:18 PM
To: Straszheim, Warren E [BIOTC]

Organization: Arizona State University

Title-Subject: [Filtered] Wet method on Evironmental SEM for cornea

Message: Hi, Everyone,

Thanks a lot for replying from several people for my previous email which asking for a way to do SEM on cornea. I have learned a lot form these replies. There are so many things I have to go through sample preparation and actually we do not have most of the instruments in our lab, but there are available in School of Biology.

I read these replies carefully and I did not see anyone who mentioned wet method on environmental SEM to get images from cornea. We have Fei
XL-30 Environmental SEM. We can do wet method.

A wet method is to introduce some water vapor near the sample to keep sample wet. The humidity can be controlled to 100%.

I think that it might be difficult to do so. On this list server there are many experts in biology field, but none of them mentioned wet method. I think if it is a good way to work on cornea, people would have told me. Many years ago I tried wet method on the surface of the samples in water. I did not get much useful information. I might miss some thing when I did it.

Therefore if someone has some experiences on wet method, even better on cornea, please tell me. It will be great.

Thanks to Stephane, Lee Cohen-Gould, Debra Sherman and Ed Haller and more people who replied to my previous email.

Zhenquan Liu
Arizona State University

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Hi, All-

I'm helping to dismantle an old EM lab on short notice. Mostly I took
pictures of stuff that others on this campus (University of Hawaii at
Manoa) might want, but I thought I'd see if anyone here needs any parts.
Small parts. I'm willing to grab and ship smaller, easy-to-pack items
only. I can be bribed with chocolate. I'm too old and cranky to ship whole
electron microscopes anymore.

You can see some pictures at
https://www.hawaii.edu/filedrop/dl/hLsCi-hIrgZ-WRbVA-WRCIZ/

There's a Zeiss DSM962 regular tungsten SEM, plugged in and pumping all
this time, but probably not used since 2009-ish. Spare parts kit and
manuals. The newest part on it is probably the penning gauge. I'm willing
to grab that sort of thing (the penning gauge).

It has an Oxford Link EDS system. Probably useless now.

There's the oldest Denton DV-502 vacuum evaporator I've ever used. Gauges
might work.

There's a BioRad PT7100 RF Plasma Barrel Etcher that I know nothing about.

And there's a Hummer II sputter coater that I used to own and was sorry I
gave to them long ago, but probably doesn't work. I didn't check for
targets, but I can.

Slim pickings, but I promised them I'd see if anyone wanted anything
before it all has to go, like by Tuesday!

Aloha,
Tina

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

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From popular_pharmacy2-at-hinet.net Thu Jun 5 00:38:44 2014
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Dear Scott,

the most important point in my view is to know if the presence of NaOH is critical for the maintenance of the structure of the NP. If not then it is not such a big problem.
NaOH can be eliminate by ultracentrifugation or filtration, provided that the filter pore size is adequate.
A note of caution however: if I am right iron is ferromagnetic and the NP may be attracted by the strong lenses of the TEM. I would be more concerned by that.

Regards,
Stephane

--------------------------------------------
On Thu, 6/5/14, zaluzec-at-microscopy.com {zaluzec-at-microscopy.com} wrote:

Subject: [Microscopy] viaWWW:In-situ NP measurement
To: nizets2-at-yahoo.com
Date: Thursday, June 5, 2014, 3:19 AM

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Email: sbras-at-uw.edu
Name: Scott Braswell

Organization: University of Washington

Title-Subject: [Filtered] In-situ NP measurement

Message: Hi,

I received an email yesterday asking me to characterize the
size of FeOx
nanoparticles in strong NaHO. I wonder if drying the
solution would
result in too much salt. My thoughts are to either try
cryoTEM or use a
liquid-cell TEM holder. Any insight you can offer for this
sample prep
would be appreciated.

Thanks,
Scott

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Fluorescent dyes which stain nucleic acids are intercalating agents, don't expect them to be "vital" (well it depends on how long you want to observe the living cells). Vital stains also have to permeate live cells and even permeate them 2x for the nucleus, one through the cell membrane and one for the nuclear envelope, which is not so easy.
I don't mean to be deceptive but as far as I know all nuclear labels are based on nucleic acid stain, meaning they are not vital.
That said, it is seldom necessary to stain the nucleus both in LM and EM in single cell experiments because it is such a big structure with high contrast.
I hope someone proves me wrote and offers you a solution.

regards,
Stephane

--------------------------------------------
On Wed, 6/4/14, jpflugheber-at-stlawu.edu {jpflugheber-at-stlawu.edu} wrote:

Subject: [Microscopy] Vital DNA stain
To: nizets2-at-yahoo.com
Date: Wednesday, June 4, 2014, 7:34 PM

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DRAQ5 is far red, so 488 excitation would be far from
optimal. What about SYBR Green?

Jill Pflugheber
St. Lawrence Universtiy

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Sent: Wednesday, June 04, 2014 12:54 PM
To: Jill Pflugheber
Subject: [Microscopy] RE: Vital DNA stain

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Have you tried DRAQ5? You could excite the dye with a 488 nm
laser - Is that what you mean "green channel"?

Zhaojie Zhang
University of Wyoming

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Sent: Wednesday, June 04, 2014 10:33 AM
To: Z.J. Zhang
Subject: [Microscopy] Vital DNA stain

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I am trying to correlate super resolution imaging with TEM
on Tokuyasu frozen sections. As an internal marker in
fluorescence, we would like to stain the the nuclei before
cryoprotection/freezing. I would appreciate any
recommendations for a rather robust vital nuclear stain. The
STED imaging is done in green channel.

Thanks, Michal
--

Michal Jarnik, Ph.D.

Electron Microscope Facility

Cell Biology and Metabolism Program

National Institute of Child Health and Human Development
National Institutes of Health

9000 Rockville Pike, Bldg. 18T, Room 101 Bethesda, MD 20892

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From medications_canadian5-at-twinmagicians.com Thu Jun 5 07:45:32 2014
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I have worked with XL30 ESEM-FEG and with Quantas. For biological
samples there are some realities often poorly understood until you
starting trying things yourself:

1) You can't really see much on samples that are truly "wet". You
will see a flat boring film of water.
2) There is a point during the pumping process, before the ESEM valve
starts cycling, where you actually have no control over the chamber
environment. Some dehydration WILL occur during this stage.
3) If samples are in solution, any dehydration will cause salts to
precipitate and obscure what you want to see.

Assuming you have the cooling stage, these problems can be (mostly)
worked around. Placing a few drops of water around the sample, on the
Peltier stage, can help keep the local environment "more humid" during
pumpdown. Pre-chilling the sample helps for the same reason. Using
too much water can lead to other problems, such as rapid bubbling
which can dislodge a loose sample or splatter on the final
lens/detector/etc. Keep a long working distance during pumpdown.

As Warren mentioned, make sure you really understand what they/you are
looking for. Growth or debris on the surface? These should be
possible. Shape distortions or internal structure? Maybe not so
much.

I did actually look at corneas and other eye parts some years ago.
The truth is that we got the best results, given what the customer was
looking for, with a 'gentle' fixative followed by a few rinses and
then imaging in ESEM. Sorry that I can't remember the fix or
solution, a med student came up with those. You are still getting
some advantage by not coating the samples, and you have several
minutes to an hour or more in which to study the surface...if your
humidity control is good. This is certainly not to say that ESEM
doesn't work, it's great for many things but you will have trouble
with the most delicate, high moisture content samples. In reality you
are trying to keep the humidity as close to 100% as you can, and work
reasonably quickly. Once water droplets actually form on your sample,
imaging is usually poor. Make small adjustments with ESEM pressure
and stage temperature, wait for the system to stabilize a bit and go
again.

Also note that the Peltier sample holders cannot be too large. We had
one made to keep the entire cornea or lens in contact with cold metal.
Even then, the top surface may be wetter OR dryer than the rest.

Make sure you watch some samples drying out so that you really
understand what the initial dehydration artifacts will look like.
Good luck!

Matt Lynn
Forensic Analytical

On Wed, Jun 4, 2014 at 8:25 PM, {wesaia-at-iastate.edu} wrote:
}
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} We have an FEI Quanta and could probably work in the same regime as you can. Also like you, I am a materials scientist, not a biologist, but I get the occasional question to look at biological material. Most recently, it was an aggregate of bacteria. The request came from a Civil/Environmental Engineer so I don't think they understood what they were asking nor would they quite know how to interpret what I found, if I had found something. That should be the first question - What are they looking for and what do they expect it to look like?
}
} A lot of biological material will not be that interesting given all the water present and the organic membranes. In my little experience, the unprepared samples just don't have much contrast when you are looking at the outside. It is cell wall and maybe a film of water. I would be interested in hearing what the biologists have to say about imparting contrast. Of course, the structure should be much more interesting on the inside but I suppose it needs enhancement, even then.
}
} I understand it is technically possible to keep a sample from dehydrating. The triple point of water is at 0.006 atm or 600 Pa at near 0C, so it should be possible to maintain a saturated atmosphere. Of course, that requires a cooling stage. If you are working at room temperature, that number rises to about 2300 Pa. That presents more of a challenge. I was not able to keep my bacteria granule from drying out in the SEM.
}
} I'll be interested in hearing any recipes that prove helpful.
}
} Warren Straszheim
} Iowa State University
}
} -----Original Message-----
} X-from: zaluzec-at-microscopy.com [mailto:zaluzec-at-microscopy.com]
} Sent: Wednesday, June 04, 2014 8:18 PM
} To: Straszheim, Warren E [BIOTC]
} Subject: [Microscopy] viaWWW:Wet method on Evironmental SEM for cornea
}
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} Email: zhenquan.liu-at-asu.edu
} Name: Zhenquan Liu
}
} Organization: Arizona State University
}
} Title-Subject: [Filtered] Wet method on Evironmental SEM for cornea
}
} Message: Hi, Everyone,
}
} Thanks a lot for replying from several people for my previous email which asking for a way to do SEM on cornea. I have learned a lot form these replies. There are so many things I have to go through sample preparation and actually we do not have most of the instruments in our lab, but there are available in School of Biology.
}
} I read these replies carefully and I did not see anyone who mentioned wet method on environmental SEM to get images from cornea. We have Fei
} XL-30 Environmental SEM. We can do wet method.
}
} A wet method is to introduce some water vapor near the sample to keep sample wet. The humidity can be controlled to 100%.
}
} I think that it might be difficult to do so. On this list server there are many experts in biology field, but none of them mentioned wet method. I think if it is a good way to work on cornea, people would have told me. Many years ago I tried wet method on the surface of the samples in water. I did not get much useful information. I might miss some thing when I did it.
}
} Therefore if someone has some experiences on wet method, even better on cornea, please tell me. It will be great.
}
} Thanks to Stephane, Lee Cohen-Gould, Debra Sherman and Ed Haller and more people who replied to my previous email.
}
} Zhenquan Liu
} Arizona State University
}
}
}
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Title-Subject: [Filtered] annealing nanoparticles on c-film grids

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Organization: University of Southern California

Title-Subject: [Filtered] Edwards 306 Turbo Evaporator

Message: Dear All,

I wonder if there are experts in working with an Edwards 306 Turbo
Evaporator.

We have had this machine working with no trouble at all for many years
but had to move it.

Once moved we re-plumbed the water cooling system and wired it up
following recommended instructions.

Now it will pump down but we cannot get power to the electrodes.
Following the instruction book we have reset all the circuit breakers we
could find but to no avail. The small LEDs do nto even light up on the
control panel next to the LT/HT switch.

Ideally it would be great to have someone we could contact in Southern
California (Pasadena area) who could take a look at the machine for us -
and get it working.

Failing that, is there anyone who could offer some useful tips for us to
try so that we can get this great machine working again.

Sincerely,

Paul Webster.

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Dear Marissa,

I have been heating C-films on gold 200 mesh grids up to 1000°C in-situ
in the electron diffraction camera without damage for the film during my
work on the lowering of the melting temperature of gold nanoparticles
(P. Buffat, Phys Rev A13 (1976) 2287). The oven was designed to create a
black body environment and ensure a nanoparticle temperature
measurement as accurate as possible.

Trials with Mo grids failed because either the grid roughness forbids
any film gliding under thermal expansion or a chemical reaction occurs
between Mo and C that could lead to Mo-carbide.

Kind regards

Philippe Buffat

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Hi Marissa,

I never encountered any issues when performing annealing experiments
using a conventional heating holder and a variety of support films.
Silicon nitride films seem robust regardless of ultimate temperature
and ramp rate. Carbon films, however, will start to show evidence of
partial graphitization at just a few hundred C, and your NPs can
become enveloped in sheets of graphitic-like carbon. This can be a big
problem if you're studying a process which requires matter transport
to or from the particle, as presumably the graphitic sheets will
impede this transport.

You also need to keep in mind that your metal grid can generate NPs.
See: Z. Zhang and D. Su, Behaviour of TEM metal-grids during in-situ
heating experiments. Ultramicroscopy 109, 766 (2009).

Good luck,
Chris
Virginia Tech

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5, 29 -- Subject: Re: [Microscopy] viaWWW:annealing nanoparticles on c-film grids
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From reasonable-meds5-at-cablebg.net Fri Jun 6 10:34:14 2014
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Email: zhangyc-at-gmail.com
Name: Yucheng Zhang

Organization: EMPA, Switzerland

Title-Subject: [Filtered] JOEL TEM heating holder

Message: Dear EM community,
I am trying to using a JEOL TEM heating holder ((Model 652) on a JEOL 2200FS TEM microsope. The
holder was originally used on a JEOL 2010 microscope but I suppose it is also compatible with the
2200FS microscope. However, when loading the holder (without heating it up), there always seems to
be a vaccum leakage in the specimen chamber(PiG4), although the vacuum of the microscope column
(PiG1) remains good. The two O-rings on the holder are checked properly and should not be the problem.
I am wondering if someone can share some experience with the holder. Is the holder really
compatible with the 2200FS microscope? If so, what could cause the vacuum crash in the specimen chamber?
Any advice would be greatly appreciated. Thanks in advance.

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Email: sunxh163-at-gmail.com
Name: Xuanhao Sun

Organization: University of Connecticut

Title-Subject: [Filtered] Manual for LKB 2088 Ultrotome V

Message: Hi,

Does anyone have a user/service manual for LKB 2088 Ultrotome V? We recently
started to see chatter in our ultrathin sections. This seems to be caused by the
instrument itself. Any advice on this matter will be appreciated!

Xuanhao Sun, Ph.D.

Bioscience Electron Microscopy Laboratory
University of Connecticut
BPB G06, Unit 3242
Storrs, CT 06269-3242

Office Tel: 860-486-6368
Lab Tel: 860-486-2914
Fax: 860-486-6369
website: emlab.uconn.edu

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Email: rich-at-tescan-usa.com
Name: Richard Fiore

Organization: Tescan USA

Title-Subject: [Filtered] NanoDay and Electron Microscopy (EM) Workshop at the University of Maryland

Message: Dear EM and FIB Friends:

I would like to take this opportunity to invite you to the NanoDay and Electron Microscopy (EM)
Workshop that will be held at the University of Maryland, College Park, Maryland on June 11, 12 and
13, 2014.

Below are some of the highlights for this meeting:

* Plenary talks on Nanoscience and nanotechnology

* EM Workshop covers a broad spectrum of newest electron microscopy and focus ion beam technology

* Platform presentations and poster session

* Live demos (both with lecture and also in the laboratory)

* Vendor exhibits

* Free registration

Additional information can be found at http://www.nanocenter.umd.edu/nanoday/

Hope to see you next week.

Best regards,

The NISP Lab
NanoCenter, UMD

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From drugs-cheapest14-at-kronos.com Sat Jun 7 08:28:38 2014
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Message-Id: {201406071328.s57DSY6q010415-at-ns.microscopy.com}

Hi
Need some advice about the effect of sputtering twice the same sample for
SEM. I had a delicate preparation (pre-embryo hatching from the zona
pellucida) showing fine details in SEM. Then I removed some rubbish from the
surface of the sample and gave it a second gold sputtering with exactly same
settings but half the time of the first one. In the second observation the
surface was like "fried" and fine details were lost. Please click here:
http://www.eikonika.net/v2/download4.php (upper foto is the first
sputtering, lower is the second)
Looks like the surface was either blebbing or it got a heavy and uneven
disposition of gold particles. I was surprised by such a strong detrimental
effect from re-sputtering and wonder if anybody knows more about this
phenomenon and how can be avoided or minimized.
Thanks a lot
yorgos

Dr Yorgos Nikas
Athens Innovative Microscopy
Skra 36 Voula 16673 GREECE

Tel/fax +30 210 8957677
mobile +30 6945 107477
www.eikonika.net www.aim.cat
************************************

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4, 22 -- From eikonika-at-otenet.gr Sun Jun 8 04:28:23 2014
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4, 22 -- From: "yorgos nikas" {eikonika-at-otenet.gr}
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4, 22 -- Subject: re-sputtering matters in SEM
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From canadian-meds2-at-bull.uk.com Sun Jun 8 06:43:24 2014
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Message-Id: {201406081143.s58Bgx80013670-at-ns.microscopy.com}

Hi Yorgos

I am not a biologist, but I have worked with all sorts of sputtering
systems, and what you see in your second picture has nothing to do with
sputter coating.

Your second picture would suggest to me that you have removed a surface
layer and you are seeing underlying structures; which is quite fascinating
but quite typical with SEM studies.

Enjoy

Steve

Steve Chapman FRMS
Protrain for Consultancy and Courses in Electron Microscopy
Mobile +44 (0) 7711 606967 Fax +44 (0)1280 814007
www.emcourses.com

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To: protrain-at-emcourses.com

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Email: marshall-at-sfu.ca
Name: Dan

Organization: Simon Fraser University

Title-Subject: [Filtered] Looking for pinouts for Oxford INCA PentaFET x3 Model 7060

Message: Hello Listers:
One of our local users has donated a Penta FET x3 to our lab. The corresponding PHA and other
collection electronics were unfortunately junked a while ago. I would like to try and marry this
PentaFET to an existing APTEC FlatPack power supply and PHA. I got very little help from Oxford and
was informed "This is all I'm allowed to give out", after some scant information on 4 of the pins.
At the moment this is what we have or have been able to figure out. If anyone can supply addditional
information on signals/volages that should be coming out or going into the Oxford EDS DB15 pin
connector it would be much appreciated.
In fact information on a similar detector would probably be a big helps as well.
Pin 1: Conditioner (15VDC used when conditioning detector?)
Pin 2: Signal Out
Pin 3: Not used
Pin 4: Not used
Pin 5: Substrate (substrate to the MOSFET, grounded?)
Pin 6: Ground?
Pin 7: Heater
Pin 8: Restore (very important pin, no info)
Pin 9: Signal return (signal ground?)
Pin 10: Not used
Pin 11: High Voltage - 500 VDC
Pin 12: Not used
Pin 13: Test Pulse (nothing known about this)
Pin 14: +24 VDC
Pin 15: -24 VDC

Any information would be much appreciated.
Thanks, Dan

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From medications-popular13-at-if.ua Sun Jun 8 22:21:58 2014
Return-Path: {medications-popular13-at-if.ua}
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Hello Steve

Thanks for your input. The problem occurs only after such delicate
biological specimens pass again from the sputter coater. Of course "after"
does not mean "because", and I keep you comment that it is not looking like
a metal disposition. Rather the specimen surface is affected and it needs a
second passage from the sputter coater for this to happen. Maybe the first
observation in SEM renders the surface susceptible to subsequent
metallization?

I use 10KV and a current beam ca 20ua that is 2-3 times lower than the
factory recommended value, because I found that these settings prolong 4-5
times the filament's life. Also my sputter coating is not optimal because I
don't use argon. Could these aberrations be involved in the problem? I think
this is possible, especially since I had no comments from people using more
orthodox settings (somebody must have tried second sputtering).

Probably the best way to tackle the issue is to use very short times of
re-sputtering and see..

Have a good day

yorgos

----- Original Message -----
X-from: "Steve Chapman" {protrain-at-emcourses.com}
To: {eikonika-at-otenet.gr}
Cc: "Microscopical Soc of America" {microscopy-at-microscopy.com}
Sent: Sunday, June 08, 2014 6:51 PM

Hello Yorgos,
It seems to me, that your samples were stored in the high humidity
before second sputter-coater run. In such conditions the biological
tissues might be rehydrated and then their surface might collapse in the
sputter-coated vacuum. This might result in structures you have seen
after second sputter-coating.

Best regards Oldrich

--
Oldrich Benada
Institute of Microbiology AS CR, v.v.i.
Videnska 1083
142 20 Prague 4
Czech Republic

On Sun, 8 Jun 2014 04:46:08 -0500, eikonika-at-otenet.gr wrote :
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} Hi
} Need some advice about the effect of sputtering twice the same sample
} for SEM. I had a delicate preparation (pre-embryo hatching from the
} zona pellucida) showing fine details in SEM. Then I removed some
} rubbish from the surface of the sample and gave it a second gold
} sputtering with exactly same settings but half the time of the first
} one. In the second observation the surface was like "fried" and fine
} details were lost. Please click here:
} http://www.eikonika.net/v2/download4.php (upper foto is the first
} sputtering, lower is the second) Looks like the surface was either
} blebbing or it got a heavy and uneven disposition of gold particles.
} I was surprised by such a strong detrimental effect from
} re-sputtering and wonder if anybody knows more about this phenomenon
} and how can be avoided or minimized. Thanks a lot
} yorgos
}
} Dr Yorgos Nikas
} Athens Innovative Microscopy
} Skra 36 Voula 16673 GREECE
}
} Tel/fax +30 210 8957677
} mobile +30 6945 107477
} www.eikonika.net www.aim.cat
} ************************************
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5, 40 -- From benada-at-biomed.cas.cz Mon Jun 9 03:59:49 2014
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Hi Again Yorgos

For some difficult samples I often have to use multiple coatings, with
different tilt angles, but only for short periods, say 30 seconds per coat.
Multiple coating may build structure on a specimen, but that is usually only
visible above 15,000X if carried out with care. In some laboratories the
operators forget that even when coated specimens may charge, and the best
solution is to lower the accelerating voltage as well as to coat. Modern
instruments work very well at 5kV or lower, and if you continue to use the
low emission currents that you mention that too helps reduce charge. The
third route to operation at a lower charge rate is to use a spot size/probe
current that is far smaller/lower than normal. For example working at 5,000X
use a probe current that would be more suitable for 30,000X; all part of the
small steps that you need to make to operate without the complexities of too
much coating. If you have a TTL instrument move away from the upper detector
to introduce more of the converted backscatter (SE3) to subdue charge.

Using air for sputtering does not make that much difference at low
magnifications, it just means you will have a different coating on a dry day
compared with a wet day. Long coating periods, particularly using old
style high voltage sputter coaters, may cook the specimen! However the
modern low voltage coaters should never reach anything like the same heat
generation.

After all that I have said there is one sure route to determining the true
"surface" structure of your specimen, and that is to use all of the tricks I
have mentioned above and very low accelerating voltage; i.e. 1 or 2kV.
Even if you are only able to look at the specimen for just a short time, it
will give you an idea of what the true structure is. But remember you are
now penetrating far less into the material so the structure may be totally
different from your 10kV!

I never look at a specimen with a sputter coating until I have tried to see
what it looks like and how it behaves using all of the low dose methods
mentioned above.

May I say I love the grey scale that you have produced with your SEM
pictures, for me the levels are very good.

Keep trying and I am sure you will see that sputter coating may not be your
problem?

Regards

Steve

Steve Chapman FRMS
Protrain for Consultancy and Courses in Electron Microscopy
Mobile +44 (0) 7711 606967 Fax +44 (0)1280 814007
www.emcourses.com

This email is intended for the person or company named and access by anyone
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prohibited and may be unlawful

-----Original Message-----
X-from: yorgos nikas [mailto:eikonika-at-otenet.gr]
Sent: 09 June 2014 07:09
To: protrain-at-emcourses.com
Cc: Microscopical Soc of America

Hi Dan,

Responding thru the listserv as your email address keeps bouncing.

I have a non-functioning Oxford Inca Pentafet detector that might be
useful in solving your pinout problem as it could be "destructively"
deconstructed to trace some of the connections. Happy to send it to
you if you wanted to pay shipping. I could probably take off the dewar
so the thing would ship in a smaller box.

Let me know,

Jim

--

James M. Ehrman
Digital Microscopy Facility
Mount Allison University
63B York St.
Sackville, NB E4L 1G7
CANADA

phone: 506-364-2519
fax: 506-364-2505
email: jehrman-at-mta.ca
www: http://www.mta.ca/dmf

By definition, one divided by
zero is undefined.

On 08/06/2014 7:27 PM, microscopylistserver-noreply-at-microscopy.com wrote:
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} Email: marshall-at-sfu.ca
} Name: Dan
}
} Organization: Simon Fraser University
}
} Title-Subject: [Filtered] Looking for pinouts for Oxford INCA PentaFET x3 Model 7060
}
} Message: Hello Listers:
} One of our local users has donated a Penta FET x3 to our lab. The corresponding PHA and other
} collection electronics were unfortunately junked a while ago. I would like to try and marry this
} PentaFET to an existing APTEC FlatPack power supply and PHA. I got very little help from Oxford and
} was informed "This is all I'm allowed to give out", after some scant information on 4 of the pins.
} At the moment this is what we have or have been able to figure out. If anyone can supply addditional
} information on signals/volages that should be coming out or going into the Oxford EDS DB15 pin
} connector it would be much appreciated.
} In fact information on a similar detector would probably be a big helps as well.
} Pin 1: Conditioner (15VDC used when conditioning detector?)
} Pin 2: Signal Out
} Pin 3: Not used
} Pin 4: Not used
} Pin 5: Substrate (substrate to the MOSFET, grounded?)
} Pin 6: Ground?
} Pin 7: Heater
} Pin 8: Restore (very important pin, no info)
} Pin 9: Signal return (signal ground?)
} Pin 10: Not used
} Pin 11: High Voltage - 500 VDC
} Pin 12: Not used
} Pin 13: Test Pulse (nothing known about this)
} Pin 14: +24 VDC
} Pin 15: -24 VDC
}
} Any information would be much appreciated.
} Thanks, Dan
}
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Has anybody tried Digital Micrograph in a Windows 8.1 environment? I have heard mixed stories, and the agent in the Microsoft store insisted that anything that runs in Windows 7 will run in 8.1.

John Mardinly, ASU

==============================Original Headers==============================
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Hi John,

I'm running DM 2.11 (actually Gatan Microscopy Suite) under Win8.1. It
runs fine. I will, however, dispute the agent's claim about ALL Win7
apps running OK under Win8.1

Cheers,
Henk

On 6/9/2014 3:45 PM, John.Mardinly-at-asu.edu wrote:
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--

Hendrik O. Colijn

*C*enter for *E*lectron *M*icroscopy & *A*nalysi*S*

1305 Kinnear Rd, Suite 100, Columbus, OH 43212

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cemas.osu.edu

"Time is that quality of nature which keeps things from happening all at
once. Lately it doesn't seem to be working."

==============================Original Headers==============================
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I was a bit hasty in my response to John. I should have indicated that
I'm running the OFF-LINE version of DM. Note that the device drivers
that talk to the hardware may or may not work under Win8.

Cheers,
Henk

On 6/9/2014 3:52 PM, colijn.1-at-osu.edu wrote:
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} Hi John,
}
} I'm running DM 2.11 (actually Gatan Microscopy Suite) under Win8.1. It
} runs fine. I will, however, dispute the agent's claim about ALL Win7
} apps running OK under Win8.1
}
} Cheers,
} Henk
}
}
} On 6/9/2014 3:45 PM, John.Mardinly-at-asu.edu wrote:
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} } Has anybody tried Digital Micrograph in a Windows 8.1 environment? I have heard mixed stories, and the agent in the Microsoft store insisted that anything that runs in Windows 7 will run in 8.1.
} }
} } John Mardinly, ASU
} }
} }
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--

Hendrik O. Colijn

*C*enter for *E*lectron *M*icroscopy & *A*nalysi*S*

1305 Kinnear Rd, Suite 100, Columbus, OH 43212

colijn.1-at-osu.edu {mailto:colijn.1-at-osu.edu} 614.643.3458 Office

cemas.osu.edu

"Time is that quality of nature which keeps things from happening all at
once. Lately it doesn't seem to be working."

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Email: bret.judson-at-bc.edu
Name: Bret Judson

Organization: Boston College

Title-Subject: [Filtered] Philips CM10

Message: To all,

We recently donated our Philips CM10 and there were some items left over
that I would like to see sent to a new home.

3 manuals: Electronics CM10/CM12 Parts 1,2, and 3 (2 manuals) and
Mechanical CM10/CM12 (1 manual)

2 exchange units and accessories.

These items are free.

Best regards,

Bret
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Organization: University of Idaho

Title-Subject: [Filtered] Asphalt samples in SEM

Message: Greetings,

Have a request to run natural asphalt samples on the SEM for imaging and EDS microanalysis.
Customer [astrobiologist] is interested framboidal pyrite as a biomarker in samples from an asphalt
lake. He describes samples as "gooey" which gives me pause.

I am in need of guidance from anyone who has experience with asphalt or tar samples on the SEM? Any
precautions, caveats, or sample prep tips.

Any assistance or recommended references would be greatly appreciated!

Tom Williams
Electron Microscopy Center
University of Idaho

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If it is gooey, expect a lot of outgassing and possible -- even probable -- contamination. Can you get him to run a specimen through a GCMS and determine what volatiles are present in what concentrations?

Mike Andrews, W5EGO
WWME Oklahoma area executive team

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} Email: tomw-at-uidaho.edu
} Name: Tom Williams
}
} Organization: University of Idaho
}
} Title-Subject: [Filtered] Asphalt samples in SEM
}
} Message: Greetings,
}
} Have a request to run natural asphalt samples on the SEM for imaging and EDS microanalysis.
} Customer [astrobiologist] is interested framboidal pyrite as a biomarker in samples from an asphalt
} lake. He describes samples as "gooey" which gives me pause.
}
} I am in need of guidance from anyone who has experience with asphalt or tar samples on the SEM? Any
} precautions, caveats, or sample prep tips.
}
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Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Tom:

1. Dissolve & Filter

Dissolve a small amount of the asphalt material in xylene (this may take an
hour or two; mix every once in awhile).

Filter the dissolved asphalt material through a steel (larger particles
50-500 um) or aluminum (smaller 0.5-20 um pore) filter. [you'll clog the
small ones quickly; but that's ok since you're looking for semi-quant data].

[Alternatively use a soxhlet extraction system]

Wash it again in xylene.

If there is any pyrite with the carbonates, you should be able to see it in
either a Reflected Polarized Light Microscopy or in the SEM.

2. Ash & Mount

Ash in muffle furnace at 350-500C for 0.5-3 hrs.

Mount the residue on a stub.

Coat for HV SEM or not for LV SEM.

3. Also, you may want to get a copy of:

Greer: Coal Microstructure & the Significance of Pyrite Inclusions, SEM, Vol
I, 79-93, 1977.

Tony

Ref: Havics, A: Microscopy of Some Bitumen-, Asphalt-, and Mastic-Based
Building Materials. Inter/Micro-2013.

.........................................
Andrew Anthony "Tony" Havics, CIH, PE
Environmental, Health & Safety, Microscopy, Materials Science & Forensic
Engineering
pH2, LLC
5250 E US Highway 36, Suite 830
Avon, IN 46123
(317) 718-7020 Office
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(317) 409-3238 Cell
aahavics-at-pH2LLC.com
www.ph2LLC.com

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Email: tomw-at-uidaho.edu
Name: Tom Williams

Organization: University of Idaho

Title-Subject: [Filtered] Asphalt samples in SEM

Message: Greetings,

Have a request to run natural asphalt samples on the SEM for imaging and EDS
microanalysis.
Customer [astrobiologist] is interested framboidal pyrite as a biomarker in
samples from an asphalt
lake. He describes samples as "gooey" which gives me pause.

I am in need of guidance from anyone who has experience with asphalt or tar
samples on the SEM? Any
precautions, caveats, or sample prep tips.

Any assistance or recommended references would be greatly appreciated!

Tom Williams
Electron Microscopy Center
University of Idaho

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From discounted_drugs13-at-webteks.com Tue Jun 10 00:17:54 2014
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Can't claim having actually done imaging of tar in SEM, but I wouldn't
dare to put asphalt into otherwise clean instrument unless there was a
way to cool it prior to pumping and keep cooled during imaging (i.e.
cryo-stage) to prevent evaporation of (and coating insides of chamber
by) heavy volatiles...

Valery Ray
=================================
PBS&T, MEO Engineering Co., Inc.
290 Broadway, Suite 298
Methuen, MA 01844 USA
Phone: +1-978-296-5063 - leave a message with call-back number
US Mobile: +1-978-305-0479
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Web: www.freudlabs.com

On 6/9/2014 7:54 PM, udsd007-at-gmail.com wrote:
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} If it is gooey, expect a lot of outgassing and possible -- even probable -- contamination. Can you get him to run a specimen through a GCMS and determine what volatiles are present in what concentrations?
}
} Mike Andrews, W5EGO
} WWME Oklahoma area executive team
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} } Name: Tom Williams
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} } Organization: University of Idaho
} }
} } Title-Subject: [Filtered] Asphalt samples in SEM
} }
} } Message: Greetings,
} }
} } Have a request to run natural asphalt samples on the SEM for imaging and EDS microanalysis.
} } Customer [astrobiologist] is interested framboidal pyrite as a biomarker in samples from an asphalt
} } lake. He describes samples as "gooey" which gives me pause.
} }
} } I am in need of guidance from anyone who has experience with asphalt or tar samples on the SEM? Any
} } precautions, caveats, or sample prep tips.
} }
} } Any assistance or recommended references would be greatly appreciated!
} }
} } Tom Williams
} } Electron Microscopy Center
} } University of Idaho
} }
} }
} }
} } Login Host: 129.101.62.35
} } Listserver Email Form V - 20120416
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Thank you for your comments. As Oldrich says, re-hydration may be an issue,
however specimens more than one year old have no signs of deterioration and
humidity is generally low in Athens. But maybe the combinatiomn of some
humidity and repeat sputterring change things. I have an old style coater
(Hummer V) using high voltage and the specimen looked rather cooked (to use
Steve's word) than anything else.
Also there were two off line comments, one from a plant morphologists saying
that some changes after re-coating occur even in the relatively hard plant
surfaces. And a material scientists sayed that using gold without argon may
etch the membrane.
Since no problem occurs in the first coating even if it is a very long one,
I feel that the combination of second sputtering plus an unknown factor are
holding the answer. If anybody of you has soft animal tissue specimens, a
gold target and can take the trouble to resputter a spare specimen, this
will be of great help to understand what is going on.
Cheers
yorgos

PS: Steve, thank you for praising my gray scales. It took me almost two
decades to understand that a low contrast micrograph contains much more
information than a high contrast one, and if you wish, this information is
easy to enhance later with a computer program

Original Message -----
X-from: "Oldrich Benada" {benada-at-biomed.cas.cz}
To: {eikonika-at-otenet.gr} ; {microscopy-at-microscopy.com}
Sent: Monday, June 09, 2014 11:59 AM

As a side note, please understand that the agent is paid to sell, not to tell the truth.
If he lies and sells more, he'll be rewarded.
If he says the truth and his sells drop, he'll get problems.

--------------------------------------------
On Mon, 6/9/14, John.Mardinly-at-asu.edu {John.Mardinly-at-asu.edu} wrote:

Subject: [Microscopy] Digital Micrograph and Windows 8.1
To: nizets2-at-yahoo.com
Date: Monday, June 9, 2014, 9:53 PM

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Has anybody tried Digital Micrograph in a Windows 8.1
environment? I have heard mixed stories, and the agent in
the Microsoft store insisted that anything that runs in
Windows 7 will run in 8.1.

John Mardinly, ASU

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Well that was a blast from the past. I started working on an SEM for Raymond T. Greer in 1977. He oversaw my masters and doctorate work. I've seen quite a few framboids in my day, but they don't often come up in discussion.

Tony's comments are good. You probably want to isolate the pyrite from the asphalt. The organics would not be all that good for your SEM. They might be difficult to image except in an VP-SEM or E-SEM. Plus, you would have a lot more material to scan through trying to find the pyrite. You might as well eliminate the organics if you can and examine what's left.

Warren

-----Original Message-----
X-from: ph2-at-sprynet.com [mailto:ph2-at-sprynet.com]
Sent: Monday, June 09, 2014 9:20 PM
To: Straszheim, Warren E [BIOTC]

Tom:

1. Dissolve & Filter

Dissolve a small amount of the asphalt material in xylene (this may take an hour or two; mix every once in awhile).

Filter the dissolved asphalt material through a steel (larger particles
50-500 um) or aluminum (smaller 0.5-20 um pore) filter. [you'll clog the small ones quickly; but that's ok since you're looking for semi-quant data].

[Alternatively use a soxhlet extraction system]

Wash it again in xylene.

If there is any pyrite with the carbonates, you should be able to see it in either a Reflected Polarized Light Microscopy or in the SEM.

2. Ash & Mount

Ash in muffle furnace at 350-500C for 0.5-3 hrs.

Mount the residue on a stub.

Coat for HV SEM or not for LV SEM.

3. Also, you may want to get a copy of:

Greer: Coal Microstructure & the Significance of Pyrite Inclusions, SEM, Vol I, 79-93, 1977.

Tony

Ref: Havics, A: Microscopy of Some Bitumen-, Asphalt-, and Mastic-Based Building Materials. Inter/Micro-2013.

.........................................
Andrew Anthony "Tony" Havics, CIH, PE
Environmental, Health & Safety, Microscopy, Materials Science & Forensic Engineering pH2, LLC
5250 E US Highway 36, Suite 830
Avon, IN 46123
(317) 718-7020 Office
(317) 718-7038 Fax
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Email: tomw-at-uidaho.edu
Name: Tom Williams

Organization: University of Idaho

Title-Subject: [Filtered] Asphalt samples in SEM

Message: Greetings,

Have a request to run natural asphalt samples on the SEM for imaging and EDS
microanalysis.
Customer [astrobiologist] is interested framboidal pyrite as a biomarker in
samples from an asphalt
lake. He describes samples as "gooey" which gives me pause.

I am in need of guidance from anyone who has experience with asphalt or tar
samples on the SEM? Any
precautions, caveats, or sample prep tips.

Any assistance or recommended references would be greatly appreciated!

Tom Williams
Electron Microscopy Center
University of Idaho

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

Dear Listers,

Just a reminder about tomorrow's webinar on image integrity. It will discuss some interesting stats re: reported cases of scientific misconduct based on published images.

{http://scientific.datacolor.com/free-new-chromacal-webinar/} http://scientific.datacolor.com/free-new-chromacal-webinar/
Wednesday, June 11, 2014, 1:00PM EDT

Retractions of papers from publication have increased tenfold between 2005 and 2011. Each incident affects the reputation of scientists, academic institutions, and public perception, which can lead to reduced funding from NIH and NSF. The role of images in publication has largely been a subtopic in Publication Ethics, yet reported cases of scientific misconduct in which images have been involved ballooned to 80% from 1989 to 2008. Images require a series of specific steps to be taken in order to protect, calibrate and create publication-ready images in research and reports. Guest presenter, Jerry Sedgewick, of Imaging and Analysis, LLC will discuss these issues in detail.

For details and registration, visit {http://scientific.datacolor.com/free-new-chromacal-webinar/} http://scientific.datacolor.com/free-new-chromacal-webinar/

Best regards,
Barbara Foster, President & Chief Consultant
Microscopy/Microscopy Education*
www.MicroscopyEducation.com

*A subsidiary of The Microscopy & Imaging Place, Inc.
7101 Royal Glen Trail, Suite A
McKinney, TX 75070
P: 972-924-5310
F: 214-592-0277

MME is currently scheduling courses for now and through Fall 2014. Call us today for a free training evaluation.

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8, 38 -- Subject: WEBINAR Reminder: How Image Integrity Impacts Your Ability to
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From drugs_affordable15-at-fibertel.com.ar Tue Jun 10 16:38:17 2014
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Email: connie-at-ultrapathimaging.com
Name: Connie Cummings

Organization: MSA

Title-Subject: [Filtered] Moving a JEOL 1011

Message: I wanted to find out if anyone knows a reliable and trustworthy moving company in NC that
would move a JEOL 1011. The move would only be less than 6 miles, it takes less than 10 minutes by
car in heavy traffic, only 2 traffic lights no matter which way you take! But it still needs a
careful and SAFE ride.
Thanks in advance for the information.
Connie

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

Hello,

I was wondering if anyone is using the K2 IND confocal attachment for
white light reflected light microscopy.
We purchased a used unit and are unable to get it working. Any help
would be greatly appreciated.

Specifically, this is where we are:

The unit has a transfer lens and a transfer lens adjustment knob.
According to the manual, there need to be the following transfer lens
adjustment steps:

----------------
1. In the CF1 position, adjust the focus to maximize the image intensity
from a reflected light sample in the eyepieces.

2. Use a screwdriver to adjust the transfer lens adjustment until the
pinholes of the stationary disc are visible.
----------------

We have a custom setup with infinity corrected Nikon lenses and a Nikon
Optiphot trinocular head.
The problem we are facing is that we can find a setting where we can see
the pinholes, but in that configuration we are unable to find any
microscope focus setting where we can clearly see the sample.

Any hints ?

regards,
Stefan

==============================Original Headers==============================
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From drugs_canadian15-at-spdop.ru Wed Jun 11 11:53:54 2014
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Email: mlj55-at-drexel.edu
Name: Michael Jablonski

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Title-Subject: [Filtered] Joint Drexel University-Sandia National Laboratory Post-Doc Position Available

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Drexel University and Sandia National Laboratory. Please see the following link for details.

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

Hi,

The K2 technology was developed by Technical Instruments of San Francisco. I'd recommend that you get in touch with them.

Good hunting!

Barbara Foster, President & Chief Consultant
Microscopy/Microscopy Education*
www.MicroscopyEducation.com

*A subsidiary of The Microscopy & Imaging Place, Inc.
7101 Royal Glen Trail, Suite A
McKinney, TX 75070
P: 972-924-5310
F: 214-592-0277

MME is currently scheduling courses for now and through Fall 2014. Call us today for a free training evaluation.

At 08:47 AM 6/11/2014, you wrote:

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

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

As many of you know for the last 3-5 years there has been at first a
discrete and now a campaign about the safety of uranyl acetate.

As the only worldwide manufacturer of uranyl acetate this has caused us
great concern since now many EM brokers are making unreasonable claims
that they have a replacement or alternative to uranyl acetate when no such
product exists.

As you all know there is a risk in using any product and uranyl acetate is
no exception.

We are preparing an educational handling and instructional manual for the
safe use of uranyl acetate in conjunction with other users in Europe and
the US.

Since the material is slightly radioactive the safety officers have gone
out of their way to condemn this product and this manual is an educational
manual to assist the worker as well as the safety officer in the proper
use and handling of uranyl acetate.

Anyone wishing to give us insight as to the positive as well as the
negative interaction with their safety officers would be welcomed.
We will not publish any names.

There has been considerable hoopla regarding anti-terrorism in Europe and
the US, let me assure everyone that you could not light a fire cracker
with uranyl acetate.

Please send any comments to abesenyo-at-ibilabs.com

Thank you for your Time
Alex Besenyo

==============================Original Headers==============================
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From drugs-best3-at-mts.ru Thu Jun 12 10:14:59 2014
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Greetings,

The 2014 Microscopy and Microanalysis meetings in Hartford are less
than 2 months away and there are still a few slots left for student
bursaries* to help with things such as providing support in the
different symposia, staffing the volunteer office, monitoring use of the
Internet Café, and helping with vendor tutorial sign-up. MSA*s
student bursary program is designed to encourage students to attend the
meetings by helping to defray some of the costs and giving them an
opportunity to meet and interact with the established microscopy
community.

Bursaries will earn $10/hour for assisting with any of the tasks
mentioned above (paid by check at the end of the meetings). There is an
added bonus of a meeting shirt and $10 cash for each morning and/or
afternoon session worked to help with meals.

For those *non-students* volunteers are also needed to help with
the above mentioned meeting activities. Although not paid on an hourly
basis as the student bursaries, volunteers do receive a meeting shirt
and the same cash allotment to help with meals. They also have the
opportunity to interact more with the microscopy community as they
assist with meeting tasks.

We can*t do it without help from you, so please consider serving as a
student bursary (or volunteer). If anyone has any questions about the
bursary/volunteer program, or would like to participate, please contact
me.

Don*t forget early registration deadline is June 23!

Amanda Lawrence
Institute for Imaging and Analytical Technologies
Mississippi State University
662-325-3019
alawrence-at-i2at.msstate.edu

*Bursaries must be members of MSA or MAS, enrolled as students at a
recognized educational institution, and have paid their registration
fee.

==============================Original Headers==============================
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From drugstore_affordable15-at-wikitravel.org Thu Jun 12 16:02:50 2014
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Well, It's empty mail bag Friday, so let me ask a question.
Wired has an article on macrophotography of inclusion in minerals. They very nice images.

http://www.wired.com/2014/06/take-a-trip-through-the-strange-worlds-within-gemstones/#x

I'm not a trained geologist, but is it possible to have carbonate inclusions in quartz?

Stay safe............
Frank

This email and any of its attachments may contain confidential information intended only for the use of the addressee(s). If the reader of this email is not the intended recipient or the employee or agent responsible for delivering it to the intended recipient, you are hereby notified that any dissemination or copying of this email is strictly prohibited. If you have received this email in error, please notify us by return email at info-at-ardl.com, permanently delete the email, and destroy any printouts. If this email contains test data and/or draft reports, you are hereby notified that only a signed original test report will contain official results, a copy of which resides in the project folder located at ARDL, Inc. Thank you. Akron Rubber Development Laboratory, Inc.

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From discounted_medications14-at-kahoe.com Fri Jun 13 07:51:37 2014
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Dear listers,

We have film holders/cassettes from two TEMs, the JEOL JEM 1200EXII and
the JEM 1400. We are doing all digital imaging and have no need to keep
them. We will be surplussing them through the government as required.
Government facilities have first dibs on these but if none reply
other educational facilities can have them for the cost of shipping.

Please contact me off line if you can use them.

Opinions and experiences related are those of Pat Connelly and do not
represent the NIH.

Pat
Patricia Stranen Connelly
NHLBI EM Core Facility
National Institutes of Health
Building 14E Room 111B
Bethesda, MD 20892-5570
301-496-3491

http://www.nhlbi.nih.gov/research/intramural/researchers/core/electron-micr
oscopy-core/index.html

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Hi all,

I was asked if it would be possible to use TEM-EDS to map locations of
Zinc ions incorporated into cellulose nano fibers (CNF). These fibers can
be quite long but are often less than 20nm in width. At this point I have
no idea as to the amount of zinc that could be incorporated into the
fibers or the distribution. Does anyone have an idea as to the spacial
resolution limits of mapping nano-scale ZnO on CNFs?

I am not sure that TEM-EDS is the way to go or if another technique, such
as AFM, would work better to confirm the presence and distribution of the
Zn on the CNFs.

Debby

Debra Sherman, Founder & CSO
DS imaging LLC
Purdue Technology Center
1281 Win Hentschel Blvd
West Lafayette, IN 47906
E-mail: debby.sherman-at-dsimagingllc.com
www.dsimagingllc.com
Mobile: 765-418-8540

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My initial thoughts are that the first back-of-the-envelope calculation I would look at is minimum mass fraction and minimum detectable mass -- see Carter & Williams, 2nd ed., chapter 36.

If the Zn loading is low and they are not agglomerated into dense clusters, the whole thing might be undetectable before spatial resolution ever comes into it.

However, if you do get good ZnO clusters (~ nms), the carbonaceous nanofibers are probably small enough and poorly scattering enough that spatial resolution will be ~ probe size, I would guess. (But run the calculation -- also Chap. 36 in CW2)

A FEG with a big X-ray detector is indicated. HAADF might do the job, too, because Zn will scatter much better than carbon.

As an example, I've mapped ~hundreds of PPM aluminum in a steel, but only because it was agglomerated into aluminum oxide precipitates, and because I used a Titan w/ChemiSTEM instrument [Parish et al., J. Nucl. Mater., V445, P. 251, 2014].

Chad

---------------------
Chad M. Parish, Ph.D.
Research and Development Staff Member
Materials Science and Technology Division
Oak Ridge National Laboratory
Phone: 1 865 574 0092
Email: parishcm-at-ornl.gov

-----Original Message-----
X-from: dsherman-at-purdue.edu [mailto:dsherman-at-purdue.edu]
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To: Parish, Chad M.

Hi all,

I was asked if it would be possible to use TEM-EDS to map locations of Zinc ions incorporated into cellulose nano fibers (CNF). These fibers can be quite long but are often less than 20nm in width. At this point I have no idea as to the amount of zinc that could be incorporated into the fibers or the distribution. Does anyone have an idea as to the spacial resolution limits of mapping nano-scale ZnO on CNFs?

I am not sure that TEM-EDS is the way to go or if another technique, such as AFM, would work better to confirm the presence and distribution of the
Zn on the CNFs.

Debby

Debra Sherman, Founder & CSO
DS imaging LLC
Purdue Technology Center
1281 Win Hentschel Blvd
West Lafayette, IN 47906
E-mail: debby.sherman-at-dsimagingllc.com
www.dsimagingllc.com
Mobile: 765-418-8540

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} I am not sure that TEM-EDS is the way to go or if another technique,
} such as AFM, would work better to confirm the presence and
} distribution of the Zn on the CNFs.

This looks to be an ideal application for the atom probe tomography.
Samples are already forming sharp tips so not much sample preparation
would be necessary - just their lift-out (which could be still
difficult). But it might be more difficult to access such machine and
its a question how it would be able to analyze this heterogeneous
sample, a combination of metal and organic material.
Have a nice day,
Tomas

--
Tomas Hrncir, Ph.D.
R&D - Physics http://www.tescan.cz
TESCAN Brno +420 530353468 | tomas.hrncir-at-tescan.cz
Libusina trida 1 | 623 00 Brno | Czech Republic

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Debbie

You have an additional problem with ZnO, in
addition to what others have said.

ZnO nanoparticles are very electron beam sensitive.
My experience is that this material damages and sputters quickly
over the entire range of 80-300 kV. You will likely
need low dose techniques which are not really useful
during XEDS analysis.

Nestor
Your Friendly Neighborhood SysOp

On 6/13/14, 7:03 PM, dsherman-at-purdue.edu wrote:
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} Hi all,
}
} I was asked if it would be possible to use TEM-EDS to map locations of
} Zinc ions incorporated into cellulose nano fibers (CNF). These fibers can
} be quite long but are often less than 20nm in width. At this point I have
} no idea as to the amount of zinc that could be incorporated into the
} fibers or the distribution. Does anyone have an idea as to the spacial
} resolution limits of mapping nano-scale ZnO on CNFs?
}
} I am not sure that TEM-EDS is the way to go or if another technique, such
} as AFM, would work better to confirm the presence and distribution of the
} Zn on the CNFs.
}
} Debby
}
} Debra Sherman, Founder & CSO
} DS imaging LLC
} Purdue Technology Center
} 1281 Win Hentschel Blvd
} West Lafayette, IN 47906
} E-mail: debby.sherman-at-dsimagingllc.com
} www.dsimagingllc.com
} Mobile: 765-418-8540
}
}
}
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The box said ...
"This program requires Win 95/98/NT or better..."
So I bought a Mac !

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

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17, 26 -- From zaluzec-at-aaem.amc.anl.gov Mon Jun 16 12:45:43 2014
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From drugs-popular10-at-byfly.by Mon Jun 16 12:55:18 2014
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A postdoctoral position is available in professor Laurence Marks Group
at Northwestern University. The successful candidate will use TEM and
STEM to study metal corrosion and oxidation. Expertise in (scanning)
transmission electron microscopy is required, and familiarity with
aberration corrected electron microscopy, metallurgy and/or
electrochemistry is preferred. Interested candidates may send
inquiries/CV to professor Laurence Marks (l-marks-at-northwestern.edu )
or Dr. Yifeng Liao (Yifeng.Liao-at-gmail.com).

Regards,

Yifeng Liao
Northwestern University

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Title-Subject: [Filtered] X-Cite LED light source

Message: Has anyone tried the "new" LED light source for fluorescence microscopy? How does it
compare to the metal halide type light source?

Thanks!

De Wood

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From medications_affordable1-at-bezeqint.net Tue Jun 17 13:51:24 2014
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A postodocoral position on Image Processing for 3D reconstruction and
correlative analysis of light and electron microscopic data sets (CLEM)
is available at the Sars International Centre for Marine Molecular
Biology in Bergen, Norway.

see http://sars.no/jobs/postdoc_hausen14Sars_01.php

regards,

--

Harald Hausen, PhD
Sars International Centre for Marine Molecular Biology
Thormøhlensgate 55
NO-5008 Bergen
Norway

e-mail: harald.hausen-at-sars.uib.no
fon: (47) 55 58 43 03

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Email: vau-at-ufl.edu
Name: Karen Kelley

Organization: University of Florida

Title-Subject: [Filtered] HPF/FS considered cryo?

Message: Is HPF/FS also considered a cryo-EM technique? Or
it is only cryo when you do everything with frozen sample?

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Email: ecdickey-at-ncsu.edu
Name: Elizabeth Dickey

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Title-Subject: [Filtered] Postdoctoral Research Position

Message: A postdoctoral research position is available beginning as soon as July 1, 2014.

The position will support a major research initiative aimed at understanding the transport behavior
and temporal evolution of interfaces in non-linear dielectrics as a function of electric-field
biasing. The research program aims to develop experimental methodologies, based on novel ex-situ and
in-situ structural, thermal, and electrical characterization techniques, to investigate spatial and
temporal phenomena that control polarization switching, leakage current, degradation, and electrical
breakdown in and around interfaces in non-linear dielectrics. The research program is coordinated
with other experimental and theoretical groups, both in the US and internationally.

The postdoctoral scholar will provide leadership in experimental efforts aimed at studying the
evolution of interface stoichiometry at metal-dielectric interfaces and understanding the
implications for electrical transport. The research will focus on ex-situ and in-situ electron
microscopy and electrical property studies, and the postdoctoral scholar will coordinate these
efforts with the partnering institutions. The research will make extensive use of the
aberration-corrected FEI TitanTM G2 scanning transmission electron microscope and other analytical
equipment within the NCSU Analytical Instrumentation Facility. Results from the research program
will be disseminated at international scientific conferences and in high-impact journals.

The ideal candidate will have knowledge of dielectric and ferroelectric materials, semiconductor
physics, and have experience in scanning transmission electron microscopy.

Please apply to this position via: https://jobs.ncsu.edu/postings/38291

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Karen,
Cryo-EM is considered quick or plunge freezing of proteins, virus, molecules
and viewing in the frozen vitrified (no ice crystals) state. These usually
have no stains so EM tomography is used after digital image capture to
reconstruct the samples 3D structure (need analysis software and some
algorithms ie. FFT). HPF/FS is where live cells and tissues are frozen
under high pressure to avoid ice crystal formation/damage. This frozen
state should retain cellular structure as it was in the living state with no
chemical fixation artifacts (shrinkage, local pH changes, selective fixation
etc). AFS which follows HPF, allows for chemical
cross-linking/stabilization at low temperatures, which does not show room
temp fixation artifacts. The idea is that under low temps (-90C) the
solvents in the AFS cocktails will substitute for the frozen water in the
sample and also carry in the fixatives. Once in place these fixatives then
work at about -50C to -60C which is artifact free to the sample (compared to
ambient or cold fixation). Cryo-ultramicrotomy is where samples are frozen
and sectioned in this state for immuno-electron microscopy labelling. All
three techniques work on the basis of low temperature stabilization (liquid
nitrogen temps) of biomolecules in the living state without ice crystal
damage, chemical fixation artifacts or extraction.

Sincerely,
Michael Delannoy
Assoc. Director Johns Hopkins SOM Microscope Facility

Email: vau-at-ufl.edu
Name: Karen Kelley

Organization: University of Florida

Title-Subject: [Filtered] HPF/FS considered cryo?

Message: Is HPF/FS also considered a cryo-EM technique? Or it is only cryo
when you do everything with frozen sample?

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9, 20 -- From prvs=239394d8c=mdelann1-at-jhmi.edu Wed Jun 18 08:37:34 2014
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Email: 12hy1-at-queensu.ca
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Title-Subject: [Filtered] Communicating problem of hot stage and computer

Message: Dear all,
We have a Tecnai Osiris TEM. We purchased a Gatan Model 652 Double Tilt Heating Holder as well. But
when we tilt the hot stage with the keyboard connected to the computer, the beta tilt can not
properly control the holder, i.e, while the tilt angel displayed in the computer is 1 degree for
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appreciate any reply.
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Dear Colleagues,

We would like to invite you to submit abstracts to the MRS Fall 2014
Symposium PP Advances in “Scanning Probe Microscopy for Multimodal
Imaging at the Nanoscale”. The symposium will cover research aimed at
understanding the spatially distinct chemical and physical makeup and
dynamic processes ongoing at solid-solid, solid-liquid, and solid-gas
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A little more clarity,
The EM tomography is done on the scope with a series of tilt angles over
different fields. These tomographs are then analysed with software to get
3D structures.

Sincerely,
Michael Delannoy

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From noreply-at-systemhat.com Thu Jun 19 13:30:47 2014
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Dear Karen,

I've worked on both single particle cryoEM, cryo tomography and HPF/FS. While the FS part (in my honest opinion) is kind of borderline cryo, I think HPF is indeed a cryo technique. Just like you plunge-freeze things into a liquid ethane or propane, you sort of do the same in HPF, except the sample is thicker. But even after freezing the sample, you quickly move it to a container where the hats/planchettes can be separated that is kept at LN2 temperature. I do count it as a cryo technique. Depending on what is done in the end with the samples (whether looked at with a FIB-SEM in a cryo stage) or cryo ultramicrotomy, or FS and microtomy. Perhaps you should ask Kent McDonald for his two cents.

Hope this helps,

Mariena

Mariena Silvestry Ramos, PhD
Senior Research Specialist
Director of Operations, CryoEM Facility
Center for Structural Biology
Vanderbilt University
Medical Center North, Suite RR1207
Nashville, TN 37232-8325
ph. 615-322-4671

________________________________________
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Title-Subject: [Filtered] HPF/FS considered cryo?

Message: Is HPF/FS also considered a cryo-EM technique? Or
it is only cryo when you do everything with frozen sample?

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Hi Karen,

} Is HPF/FS also considered a cryo-EM technique? Or
} it is only cryo when you do everything with frozen sample?

there is some consensus on the following terminology:

* Cryo-TEM = Microscopy of frozen hydrated specimens at low temperature (including, but not necessarily electron tomography) that were prepared by immersion freezing, high pressure freezing, etc.

As HPF/FS specimens are being inspected at room temperature in the TEM, this is NOT cryo-TEM, but a cryo-preparation technique for TEM. The same applies, for example, to Tokuyasu cryo-sections that are later thawed for immunolabelling.

Hope that helps,

Guenter

--
Dr. Guenter Resch
Nexperion e.U. - Solutions for Electron Microscopy - www.nexperion.net
Mattiellistrasse 3/17, 1040 Vienna, Austria - Phone +43 664 94 17 210
Registered at Commercial Court Vienna, FN 397677w - VAT Reg. ATU67962234

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From drugs_express9-at-reelinfish.com Fri Jun 20 03:25:52 2014
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Researchers from another campus want to be able to prepare TEM specimens of their nanoparticles by themselves and are asking for a protocol. I want to make things a bit easier for them by substituting uranyl acetate with phosphotungstic acid for negative staining. Can something go wrong because of this replacement?

Thanks,

Vladimir

Vladimir M. Dusevich, Ph.D.
Electron Microscope Lab Director
(816) 235-2072
371 School of Dentistry
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From drugs_express1-at-global-ip.net Sat Jun 21 02:54:11 2014
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Hi Vladimir,

} Researchers from another campus want to be able to prepare TEM specimens of their nanoparticles by themselves and are asking for a protocol. I want to make things a bit easier for them by substituting uranyl acetate with phosphotungstic acid for negative staining. Can something go wrong because of this replacement?

from my own experience with UA (acidic) and PTA (neutral; usually both at 2.0%) for negative staining of viruses and macromolecular complexes, I have observed differences in stain distribution, resolution and - interesting enough - also the optimum concentration of the specimen. UA typically requires a less concentrated sample for optimal results.

Having said that, both stains can produce very nice results, but a switch from one to the other might require adjustment of some parameters in the protocol.

Best regards,

Guenter

P.S: Do these NPs actually need to be stained? Depending on the material/size/question at hand you might try to dry them down on a C film, with or without washing.

--
Dr. Guenter Resch
Nexperion e.U. - Solutions for Electron Microscopy - www.nexperion.net
Mattiellistrasse 3/17, 1040 Vienna, Austria - Phone +43 664 94 17 210
Registered at Commercial Court Vienna, FN 397677w - VAT Reg. ATU67962234

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From medications-express7-at-visualinvestments.com Sun Jun 22 01:55:47 2014
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Hello everybody!
I thought that the 2d-FFT of a HRTEM image taken in MAG mode of my TEM equals the diffraction pattern which I also could have taken in DIFF mode of the same sample area. Right?
Well, if I acquire the diffractogram in DIFF mode, then I know which camera length I have had in use. But what´s about the 2d-FFT which was calculated from a real space image? How do I know which "camera length" is represented in there?
I am asking, because I would like to save a 2d-FFT as an image and underlay it to a calculated powder diffraction pattern. But in order to scale my 2d-FFT "image" and the powder diffraction ring pattern "image" appropriately, the camera length would have to be known and I am missing how to derive that value.
Thanks,
Marco

This e-mail is from CIC biomaGUNE. The e-mail and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. Any unauthorised dissemination or copying of this e-mail or its attachments, and any use or disclosure of any information contained in them, is strictly prohibited and may be illegal. If you have received this e-mail in error, please notify or telephone + 34 943 00 53 00 and delete it from your system.

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Contents Retrieved from Microscopy Listserver Archives
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Hi Marco,

The FFT of the image isn't exactly like a diffraction pattern since the
phase information has been lost in the image. It is more properly
called a "power spectrum".

The "camera length" of the FFT is based on the image mag. The easiest
way to determine this is to take the pixel calibration of the image and
invert the value. So, if the image calibration is 2 pixels/nm or 0.5
nm/pixel, then the FFT will have a calibration of 2.0 nm-1 per pixel.

You can take your "powder" pattern, calculate the d-spacing of the rings
and from that determine the calibration of your pattern in nm-1. At
that pont, you just need to scale the two patterns identically.

Cheers,
Henk

On 6/22/2014 8:25 AM, mmoller-at-cicbiomagune.es wrote:
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} Hello everybody!
} I thought that the 2d-FFT of a HRTEM image taken in MAG mode of my TEM equals the diffraction pattern which I also could have taken in DIFF mode of the same sample area. Right?
} Well, if I acquire the diffractogram in DIFF mode, then I know which camera length I have had in use. But what´s about the 2d-FFT which was calculated from a real space image? How do I know which "camera length" is represented in there?
} I am asking, because I would like to save a 2d-FFT as an image and underlay it to a calculated powder diffraction pattern. But in order to scale my 2d-FFT "image" and the powder diffraction ring pattern "image" appropriately, the camera length would have to be known and I am missing how to derive that value.
} Thanks,
} Marco
}
}
} This e-mail is from CIC biomaGUNE. The e-mail and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. Any unauthorised dissemination or copying of this e-mail or its attachments, and any use or disclosure of any information contained in them, is strictly prohibited and may be illegal. If you have received this e-mail in error, please notify or telephone + 34 943 00 53 00 and delete it from your system.
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--

Hendrik O. Colijn

*C*enter for *E*lectron *M*icroscopy & *A*nalysi*S*

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"Time is that quality of nature which keeps things from happening all at
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17, 46 -- image?
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THANK YOU TO EVERYBODY FOR THE VERY VALUABLE INFORMATION!
Marco

On 6/22/2014 8:25 AM, mmoller-at-cicbiomagune.es wrote:
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} Hello everybody!
} I thought that the 2d-FFT of a HRTEM image taken in MAG mode of my TEM equals the diffraction pattern which I also could have taken in DIFF mode of the same sample area. Right?
} Well, if I acquire the diffractogram in DIFF mode, then I know which camera length I have had in use. But what´s about the 2d-FFT which was calculated from a real space image? How do I know which "camera length" is represented in there?
} I am asking, because I would like to save a 2d-FFT as an image and underlay it to a calculated powder diffraction pattern. But in order to scale my 2d-FFT "image" and the powder diffraction ring pattern "image" appropriately, the camera length would have to be known and I am missing how to derive that value.
} Thanks,
} Marco

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From affordable-meds6-at-zaural.ru Mon Jun 23 00:50:33 2014
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Hello Vladimir,
I agree with Guenter that you do not need to negatively stain
the NPs in some cases.
However, if NPs are conjugated with some ligands or covered with
organic layer(s), then the negatives staining is necessary. In this
case, we are routinely using ammonium molybdate in concentration ranging
from 0.1% to 1% in ddH2O. In our hands, both uranyl acetate and
phosphotungstic acid are too electron opaque and can hide the NPs.

Best regards Oldrich

--
Oldrich Benada
Institute of Microbiology AS CR, v.v.i.
Videnska 1083
142 20 Prague 4
Czech Republic

On Sat, 21 Jun 2014 10:38:40 -0500, lists-at-nexperion.net wrote :
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} Hi Vladimir,
}
} } Researchers from another campus want to be able to prepare TEM
} } specimens of their nanoparticles by themselves and are asking for a
} } protocol. I want to make things a bit easier for them by
} } substituting uranyl acetate with phosphotungstic acid for negative
} } staining. Can something go wrong because of this replacement?
}
} from my own experience with UA (acidic) and PTA (neutral; usually
} both at 2.0%) for negative staining of viruses and macromolecular
} complexes, I have observed differences in stain distribution,
} resolution and - interesting enough - also the optimum concentration
} of the specimen. UA typically requires a less concentrated sample for
} optimal results.
}
} Having said that, both stains can produce very nice results, but a
} switch from one to the other might require adjustment of some
} parameters in the protocol.
}
} Best regards,
}
} Guenter
}
} P.S: Do these NPs actually need to be stained? Depending on the
} material/size/question at hand you might try to dry them down on a C
} film, with or without washing.
}
} --
} Dr. Guenter Resch
} Nexperion e.U. - Solutions for Electron Microscopy - www.nexperion.net
} Mattiellistrasse 3/17, 1040 Vienna, Austria - Phone +43 664 94 17 210
} Registered at Commercial Court Vienna, FN 397677w - VAT Reg.
} ATU67962234
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Debby,
Thank you very much for reminding me about glow discharge (somehow it skipped away from me... )

Thanks to everybody for all replies.
Particles are organic (pharma).

Vladimir

Vladimir M. Dusevich, Ph.D.
Electron Microscope Lab Director
(816) 235-2072
371 School of Dentistry
650 E. 25th Street
Kansas City, MO 64108-2784

-----Original Message-----
X-from: Sherman, Debra [mailto:dsherman-at-purdue.edu]
Sent: Friday, June 20, 2014 3:46 PM
To: Dusevich, Vladimir

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Although most members of this listserver are hard-core microscopy
professionals, some of your junior colleagues could benefit from
attending Fourth Short Course on FIB technology and operation that will
be taking place at University of Maryland at College Park, MD on 06/30 -
07/03.

Program covers basic principles of FIB, physical sputtering, Gas
Assisted Etching/Deposition processes, and brief overview of practical
applications. The course is open to all participants - please pass the
information around and feel free to contact if you have any questions or
would like to attend.

http://nanocenter.umd.edu/events/index.php?mode=4&id=9466

--
Valery Ray
=================================
PBS&T, MEO Engineering Co., Inc.
290 Broadway, Suite 298
Methuen, MA 01844 USA
Phone: +1-978-296-5063 - leave a message with call-back number
US Mobile: +1-978-305-0479
Skype: pbstmeo
E-Mail: vray-at-partbeamsystech.com
Web: www.partbeamsystech.com
Web: www.freudlabs.com
UMD E-Mail: vray-at-umd.edu

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Hi Marco,

Others have already touched on the fact that FFT transforms of an HR image will have different phase information that the direct diffraction. Â I want to also mention that if you are computing your powder pattern using software designed for X-ray diffraction, it will usually exclude reflections which have a structure factor of zero. Â Many times, these reflections are visible in HR FFT transforms (and even in SAED diffractions). Â Therefore, you can expect to find more reflections in your HR FFT than you may guess. Â Also, expect the intensities to be very different from a calculated value.

In practical terms this can be very useful. Â Often the structure factor for large d-spacings is zero (e.g. 100 reflections are often not visible in X-ray diffraction) but these can show up in HR images. Â Since the d-spacings are large, they are usually much better discriminators of the crystal than higher order reflections.

Zack Gainsforth
Space Sciences Laboratory, UC Berkeley
7 Gauss Way
Berkeley, CA Â 94720
cell: 626-437-9186
zackg-at-ssl.berkeley.edu

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Reply: mmoller-at-cicbiomagune.es mmoller-at-cicbiomagune.es(mailto:mmoller-at-cicbiomagune.es)

Hi Vladimir

This conversation brought to mind a recent experience we had here with silver-based nanoparticles.
We don't have a specialised instrument for glow discharge, but we use a brief low KV, low amp pulse from our sputter coater (which we normally use for SEM sample coating). For most negative staining and other dried down samples, this works well.

However, for small nanoparticles (~10-20 nm diameter range) there is a problem. This procedure produces spots which are in that range and cannot be easily distinguished from nanoparticles. We know this because we always do controls. In our case we found it was not problem to examine these nanoparticles on grids which we had not treated with glow discharge. For the silver particles there was no need for negative stain of course.

I guess what I am saying is with nano-particles, make sure you do controls because there are numerous ways of particle-like artefacts of the sample prep process.

Good luck with it.

Duane

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Sent: Tuesday, 24 June 2014 3:44 a.m.
To: Harland, Duane

Debby,
Thank you very much for reminding me about glow discharge (somehow it skipped away from me... )

Thanks to everybody for all replies.
Particles are organic (pharma).

Vladimir

Vladimir M. Dusevich, Ph.D.
Electron Microscope Lab Director
(816) 235-2072
371 School of Dentistry
650 E. 25th Street
Kansas City, MO 64108-2784

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Message: I was wondering if anyone had an old Gatan model 656 dimpler that was no good any more.
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Title-Subject: [Filtered] Reichert Jung ultracut E spares

Message: Hi to all,

Recently my ultramicrotome Reichert Jung ultracut E had a problem in the motor that moves the sample
forward during sectioning, as expected, regarding the age, this motor is hard to find and probably
impossible to replace by a new one (the Berger/Lahr company no longer exists), so I am looking for a
used motor (hope you can see it in the link below) the reference is: Berger/Lahr, W-germany,
RDM253/50, IW= 240MA,RW= 37.5º. Does anyone has or know someone, or some company that has/sells
(second handed) this motor? like maybe an old ultramicrotome (same brand and model)with spare parts
to share.

https://www.flickr.com/photos/125656322-at-N02/14494669172/

Thank you so much in advance,

Hope you all have a lovely day

Andreia

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Structure Probe, Inc. is seeking an electron microscopist to work in its West Chester, PA analytical laboratory. We are a materials science oriented facility providing analytical services in the electron microscopy field to our clients on a contract basis. This individual should possess TEM skills including microtomy capabilities. BS or 5-years experience preferred. Duties include sample preparation, obtaining analytical data, working with clients and report preparation.
Further information may be found on our website at: http://www.2spi.com/employment-opp.html

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Email: dan.fairweather-at-delphi.com
Name: Dan Fairweather

Organization: Delphi Automotive, Powertrain Div

Title-Subject: [Filtered] Hitachi S3200-N variable pressure control

Message: This morning I experienced a situation on our SEM that hasn't occurred before. While
operating in variable pressure mode, the system will not control below 100 Pa. I normally operate -at-
30 Pa for non-conductive samples. It is an older system and the air supply lines are starting to
fail. But I don't hear any air leaks when I remove the front of the cabinet. We did verify the fluid
levels in our roughing pumps and topped them both.

We do have a call in to Hitachi service, but they can't get to us until July. Does any one have
experience with this problem? Are there some suggestions you can make to diagnose the problem?

Thank you in advance,
Dan Fairweather
Senior Mateials Development Engineer
Delphi Automotive

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We had a S-2460N for many years. In fact, we just shipped it off working a week ago. I think it may have had a similar problem.

About every six months to a year, the system would stop controlling and we would find that the needle valve that determines the chamber pressure had bound up. Under the original setup, the valve would never really settle down. It remained in an oscillating mode turning a half turn this way then a half turn back. The valve was always in motion. Eventually the threads galled and it became hard to turn. You could still turn it by hand - with difficulty, but the motor was not up to the task. Hitachi eventually got it figured out and changed some components in the control board so that it did settle in and the valve turned much less and lasted much longer.

Our valve was located in the right rear of the column cabinet near the top. We could remove the back panel and watch the valve in action. As I recall, when we had problems, we would unplug the motor cable and set the valve by hand for the desired pressure. I think there was another line used to vent the chamber so we could just about "set it and forget it". Of course, Hitachi then came in, replaced the valve, and we were back on our regular routine.

Contact me if you want more details.

Warren

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Email: dan.fairweather-at-delphi.com
Name: Dan Fairweather

Organization: Delphi Automotive, Powertrain Div

Title-Subject: [Filtered] Hitachi S3200-N variable pressure control

Message: This morning I experienced a situation on our SEM that hasn't occurred before. While operating in variable pressure mode, the system will not control below 100 Pa. I normally operate -at-
30 Pa for non-conductive samples. It is an older system and the air supply lines are starting to fail. But I don't hear any air leaks when I remove the front of the cabinet. We did verify the fluid levels in our roughing pumps and topped them both.

We do have a call in to Hitachi service, but they can't get to us until July. Does any one have experience with this problem? Are there some suggestions you can make to diagnose the problem?

Thank you in advance,
Dan Fairweather
Senior Mateials Development Engineer
Delphi Automotive

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From cheapest-meds4-at-dalacom.kz Wed Jun 25 23:37:43 2014
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Hi all,

I know this has been discussed before, just wondered if there were any new
thoughts. We're moving twice, first into a refurbished basement, second
time into a new building. The basement has a sprinkler system in case of
fire, unlike our current building which just has alarms (it's a single
storey brick building). I imagine the new building will have some sort of
sprinkler system too.

What precautions, if any, do people take to protect against the unlikely
showering of your confocal or EM?
Are there any relatively simple alternatives? (One alternative is to have
fire-doors and walls for each room but that is prohibitive.)

thanks much,
Rosemary

Dr Rosemary White
CSIRO Plant Industry (for 4 more days only)
GPO Box 1600
Canberra, ACT 2601

T 02 6246 5475
F 02 6246 5334
E rosemary.white-at-csiro.au

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Hi Rashmi,

} The chiller of FEI HRTEM F30 is consistently at 10 deg C. Even after increasing the tempt. it is not
} increasing. I would like to know the possible reasons: is it due to the faulty temp. controller or
} sensor?

did you measure the 10 degrees yourself or is this what is displayed on the cooling unit? This might help to identify if the sensor is broken or not.

Depending on manufacturer, another culprit might be the valve that controls the flow of water of the secondary cooling circuit through either the heat exchanger or the bypass - if that is stuck, you are constantly cooling.

Did you already try to contact the manufacturer of the cooling unit?

Best,

Guenter

--
Dr. Guenter Resch
Nexperion e.U. - Solutions for Electron Microscopy - www.nexperion.net
Mattiellistrasse 3/17, 1040 Vienna, Austria - Phone +43 664 94 17 210
Registered at Commercial Court Vienna, FN 397677w - VAT Reg. ATU67962234

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If your chiller is from Haskris, you can call them for "over the phone diagnostics". I have done it already twice; bad parts were correctly identified, ordered, and replaced by myself (my chillers are old...)

Vladimir

Vladimir M. Dusevich, Ph.D.
Electron Microscope Lab Director
(816) 235-2072
371 School of Dentistry
650 E. 25th Street
Kansas City, MO 64108-2784

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Name: Rashmi

Organization: DBT

Title-Subject: [Filtered] Chiller problem

Message: Hi!

The chiller of FEI HRTEM F30 is consistently at 10 deg C. Even after increasing the tempt. it is not increasing. I would like to know the possible reasons: is it due to the faulty temp. controller or sensor?

Thanks and Regards
Rashmi

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Dear EM slot grid users
I had some experience making this support - membrane - film for slot
grid with formvar or butvar (pioloform).
But as you may know this more an art than a highly reproducible protocol...
Very briefly:
For the one who experienced to make such membrane there are two main
critical steps:
First critical step is cleaning the glass slide
with kimwipe then with lens tissue
or
with 70% ethanol then kimwipe then with lens tissue
or
dip in alconox saturated solution then without rincing kimwipe then with
lens tissue

Casting the film on the glass slide
put the cleaned glass slide in a borel glass container half filled with
your solution of formvar or butvar.
Instead of a borel I use a homemade film casting device resembling the
one of EMS with two part glass allowing a controlled flux from the
upper part containing the glass slide to the lower container with a
special screw: "A built-in fine-pressure-release valve helps control the
speed of drainage." EMS 71305-01
I used both dissolved around 0.30-0.60% butvar in chloroform (high
quality EMSURE from Merck stabilized with 0.6% ethanol)

Detaching the film on a large and deep glass container
Some people use a blade to score the film few millimeter parallel to the
long side of the glass slide and at its bottom, some
rub the 3 side of the glass slide against a kimwipe.

Some people insert the glass slide in the water with a 90Â° angle
allowing the detachment of the membrane from the two sides of th glass
slide. Some insert it with a 45Â° angle detaching only the upper
membrane. In my hands both work well.

I have many questions regarding this procedure.
How do you keep your solution of butvar ? at 4CÂ° ? at RT ? and for how
long do you keep it ?
As you know the problem is the humidity in the room when your cast your
membrane:
I have read that some people do not cast when the air is humid !!! do
you have a way to control this ?
I tried to put some nusiev bead in the butvar solution the membrane
detached well but I haven't check the quality of the membrane yet. Do
you have experience with this ?

I should definitely have some feed back on this Oh so delicate issue

Thank you very much
Jean-Michel Hermel

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jmhermel-at-inaf.cnrs-gif.fr

Dear Jean-Michel,
congratulations for summing up all these important steps in mounting films to grids. Thank you!
A similar MSA thread has been presented / started July 18, 2012, entitled: ("[Microscopy] formvar grids" by Nicki Watson].
Unfortunately I have no experience with butvar-, but with formvar-films.

In your summary, for:

{Some people insert the glass slide in the water with a 90Â° angle allowing the detachment of the membrane from the two sides of the glass
slide. Some insert it with a 45Â° angle detaching only the upper membrane. In my hands both work well.}

I would like to add that some people try {breathing} to the glass slide with the film before inserting the slide into the water because this could help in easier detachment of the film. In my experience inserting angles between 45 and 85 degr. will work. I sometimes was able, to detach the formvar film from BOTH sides of the glass slide inserting them at really 90 degr.

Regarding your questions:
Humidity: if the air in the working room is too humid DURING FILM application, you will get holes in your film (but this depending also on the apparatus used for application of formvar.... unfortunately I could not find any German translation for {BOREL} glass container, so I cannot comprehend what / how you are doing films with such one.

I am using a self-made film casting device (principally very similar to the EMS Film Casting device you described below (cf: https://www.emsdiasum.com/microscopy/products/grids/accessories.aspx and scroll down to the middle of the display in the window).

For such a device it needs therefore primarily a formvar or butvar (pioloform) organic diluent which
i) is 100% anhydrous (i.e. either freshly made) or (since as with the EMS-model: {The film casting solution can be used repeatedly} )
ii) the possibility of a closed / totally closable/sealable system during storage of the Formvar/Butvar-solution for a longer time.

During the making of film (application to the cleaned glass slide) you normally will have arid conditions if you don't insert a humid or moist glass slide, which - if applied consecutively several times - certainly (w)could add moisture to the (anhydrous) organic solution which in turn produced "holey" films (this was -BTW - an instruction I was given by a famous ZEISS application rep. in 1981 when I started my EM-career, namely: to direct a short puff of "moisty" breath onto the glass slide receptaculum or the glass slide itself at the time the Formvar/Butvar solution completely sank down within the casting cylinder below the lower glass slide edge).
Control for moisture (i.e. {dry} environment during the film application process in the cylinder -
i) if this is an open system "upwards" from where you place the object slide for film application): mounting a phiole (can be made from glass or even plastic, the latter can be purchased as "test tubes" with both sided detachable "olives" ) containing granulate of moisture absorbing material (some kind of silica gel containing a color-indicator)
ii) the same applies for the Air-in and Air-out atomizer ball (in between ball and nozzle linking to the {drainage neck} of the apparatus)
Such construction will help in keeping the organic solution as "dry/anhydrous" as possible.

Storage of the device/solution when not in operation:
i) shield from light all time when not using the apparatus [fitting cardboard box, eventually optimized by own hands on...(:-))]
ii) storage at ambient (Room) Temperature (not in refrigerator since you will get moisture in it when "warming up to working temperature)
iii) shelf life of solution: I have made the experience that one can use a more concentrated stock solution in really (99.9%) anhydrous organic solution (e.g. Chloroform UVASOL [Methane trichloride, Trichloromethane] for more than a year, diluted solutions from that - if stored as working solution properly after dilution (light shielding and system firmly closed), for at least 1 year too.
Other diluents, like ethylene dichloride [1,2-Dichloroethane; Ethylene chloride; Glycol dichloride] or
Dioxane [Diethylene dioxide; Diethylene ether; Dioxan; p-Dioxane] I don't have an experience.

Eventually cf: Work-Study Student Handbook, Chapter 7. Grid Preparation Methods:
-at- http://metz.rose2.brandeis.edu/emdoc/books/workstudystudents-handbook/grids.html
esp. cf: http://metz.rose2.brandeis.edu/emdoc/books/workstudystudents-handbook/grids-coated.html as well as PREV & NEXT pages!
esp. cf: Making Formvar Grids in High Humidity: http://metz.rose2.brandeis.edu/emdoc/books/workstudystudents-handbook/grids-humidity.html

One question left: what did you mean when writing:
{I tried to put some { {'nusiev'} } bead in the butvar solution} ? is this a kind of molecular sieve??

By the way I have found a "classic, ancient" literature reference for Collodion/Amylacetat and Formvar/chloroform filmed Grids:
SPECIMEN PREPARATION FOR ELECTRON MICROSCOPY, by J.A. GARD, 1955, 5 pages (http://www.minersoc.org/pages/Archive-CM/Volume_3/3-15-14.pdf)

Best wishes and regards,
Wolfgang

Wolfgang MUSS
MSA-member
SALZBURG, Austria

Von: jmhermel-at-inaf.cnrs-gif.fr [mailto:jmhermel-at-inaf.cnrs-gif.fr]
Gesendet: Donnerstag, 26. Juni 2014 17:39
An: MuÃŸ Wolfgang
Betreff: [Microscopy] support membrane film for slot grid

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Dear EM slot grid users,
I had some experience making this support - membrane - film for slot grid with formvar or butvar (pioloform).

But as you may know this more an art than a highly reproducible protocol...
Very briefly:
For the one who experienced to make such membrane there are two main critical steps:

First critical step is cleaning the glass slide with kimwipe then with lens tissue
or
with 70% ethanol then kimwipe then with lens tissue
or
dip in alconox saturated solution then without rincing kimwipe then with lens tissue

Casting the film on the glass slide
put the cleaned glass slide in a borel glass container half filled with your solution of formvar or butvar.
Instead of a borel I use a homemade film casting device resembling the one of EMS with two part glass allowing a controlled flux from the
upper part containing the glass slide to the lower container with a special screw: "A built-in fine-pressure-release valve helps control the
speed of drainage." EMS 71305-01
I used both dissolved around 0.30-0.60% butvar in chloroform (high quality EMSURE from Merck stabilized with 0.6% ethanol)

Detaching the film on a large and deep glass container
Some people use a blade to score the film few millimeter parallel to the long side of the glass slide and at its bottom, some rub the 3 side of the glass slide against a kimwipe.

Some people insert the glass slide in the water with a 90Â° angle allowing the detachment of the membrane from the two sides of the glass
slide. Some insert it with a 45Â° angle detaching only the upper membrane. In my hands both work well.

I have many questions regarding this procedure.
How do you keep your solution of butvar ? at 4CÂ° ? at RT ? and for how long do you keep it ?

As you know the problem is the humidity in the room when your cast your membrane:
I have read that some people do not cast when the air is humid !!! do you have a way to control this ?

I tried to put some nusiev bead in the butvar solution the membrane detached well but I haven't check the quality of the membrane yet. Do
you have experience with this ?

I should definitely have some feed back on this Oh so delicate issue

Thank you very much
Jean-Michel Hermel

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From drugstore-popular5-at-kurkino.net.ru Thu Jun 26 20:00:00 2014
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Hi Rosemary

Out of interest I have dealt with water cascading over microscopes, yes it
has happened twice in my 49 electron microscopy years. As a service
technician the main problem is not so much the water, but the debris that
the water beings down onto the electrical circuits. Both of the instruments
in question were running on vacuum only and managed to keep going overnight
until security found the water running through the particular laboratories.
One was a tap exploding, shooting a fountain of water into the EM room, the
other the result of putting out a fire three levels above.

A third situation occurred when a water pipe came off one instrument in a
basement EM unit, flooding the complete area to about 6 inches. Everyone of
the instruments kept going, but as I arrived a technician with rubber boots
and a long wooden pole was taking care of switching the most endangered
instruments off. However this was in the 1980s when most EM Units were
underground; the one I mentioned was also used as a Tornado shelter!

EM Units and water, great fun!

Steve

Steve Chapman FRMS
Protrain for Consultancy and Courses in Electron Microscopy
Mobile +44 (0) 7711 606967 Fax +44 (0)1280 814007
www.emcourses.com

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Email: joseph.uknalis-at-ars.usda.gov
Name: Joe Uknalis

Organization: ERRC - USDA

Title-Subject: [Filtered] new sputter coater recommendation

Message: Hi folks-
Looking for a new sputter coater-

considering:

EM Sciences EMS150R

Denton Standard Desk V HP

Leica EM ACE200

or any other you may prefer.

please contact me off list.

thanks

Joe

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Email: andrei.kolmakov-at-nist.gov
Name: Andrei Kolmakov

Organization: NIST

Title-Subject: [Filtered] coater

Message: Dear Colleagues,
We are considering purchasing table-top high vacuum coater as a general user friendly instrument
which would allow :
1. sputter coating (exchangeable targets)
2. carbon coating
3. thermal and/or e-beam metal evaporation
4. glow discharge treatment and/or cleaning
We are considering Leica Microsystems AC600 and Electron Microscopy Sciences EMS 150T ES systems.
Your comments/experiences on their performance would be a great help.
Thank you in advance
Andrei

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Email: jjmccarthy-at-wisc.edu
Name: Jon McCarhty

Organization: College i=of Engineering, UW Madison

Title-Subject: [Filtered] Research Facilities Director Posting

Message: HI all;

WITH MY IMPENDING RETIREMENT, MY POSITION IS NOW POSTED.
Please read and submit if interested. Deadline is coming up on July one, but an email to Susan
Hagness with expressed interesT is advised to get an extension!

Best to all, and see you at IUMAS/MAs!

Jon McCarthy, Past MAS President
_____________________________________________
Vacancy Listing #79754 to:
Susan Hagness
Phone: 608-265-5739

1415 Engineering Dr
2620 Engineering Hall
Email: hagness-at-engr.wisc.edu

Working Title: RESEARCH USER FACILITIES DIRECTOR
Official Title: SENIOR SCIENTIST OR ASSOCIATE SCIENTIST

Degree and area of specialization:
Ph.D. in Engineering or the Physical Sciences with extensive knowledge in the field of scientific
instrumentation, microscopy, research programs and specialized research equipment.

Minimum number of years and type of relevant work experience:
Five years of experience in scientific and research environments, including the private sector,
national laboratories, and/or higher education. Experience also includes successful program
development, demonstrated knowledge and use of advanced characterization tools or micro-fabrication
technologies and managing budgets and personnel. Candidates should have a demonstrated commitment to
excellence in scientific leadership.

Position Overview
The Research User Facilities Director will report to the Associate Dean for Research and Graduate
Affairs and will be responsible for all aspects of managing the College of Engineering research user
facilities which include, but are not limited to, the Materials Science Center, Soft Materials
Laboratory and the Wisconsin Center for Applied Microelectronics.

Principal Duties:
Â• Construct a business plan for each research user facility that includes development of annual
budgets, establishment of user rates, anticipated customer base and customer needs, staffing
requirements, advisory boards, equipment inventory and capital budgets.

Â• Work with facility users and College faculty to identify and obtain new equipment as needed,
typically through participation in the development of equipment grant proposals.

Â• Maintain facility equipment, negotiating and overseeing service contracts and equipment repairs as
needed. Provide the College with an annual plan for equipment upkeep, rotation and the purchase of
new equipment.

Â• Supervise facility staff and recommending staffing changes as needed.

Â• Oversee and manage the daily operations of all research user facilities.

Â• Assist the Associate Dean for Research and Graduate Affairs in the review of faculty requests for
major equipment purchases and the transition (sale, movement, retirement) of any facility or faculty
research equipment within the College.

Â• Assist the Associate Dean for Research Administration and Policy with the review and approval of
Facility Use Agreements with industry or other outside entities.

Employee Class: Academic Staff
Department: College of Engineering DeanÂ’s Office
Full Time salary rate: Minimum $90,000 Annual (12 month).
Appointment Percent: 100%
Anticipated begin date: September 1, 2014
TO ENSURE CONSIDERATION

Application must be received by: JULY 01, 2014
HOW TO APPLY:
Please submit a cover letter and resume or CV to Patricia Droes at: pdroes-at-engr.wisc.edu
Unless another application procedure has been specified above, please send resume and cover letter
referring to Position Vacancy Listing #79754 to:
Susan Hagness
Phone: 608-265-5739

1415 Engineering Dr
Fax: N/A

2620 Engineering Hall
Email: hagness-at-engr.wisc.edu

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X-from: loisbielefeld-at-gmail.com ()

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Email: loisbielefeld-at-gmail.com
Name: Lois Bielefeld

Organization: Photographer/Artist

Title-Subject: [Filtered] Artist project video on Chromosomes assistance needed

Message: Hi,
I'm a working artist in Milwaukee, WI and am working on a project specifically about gender. I'm
looking to do a side-by-side video piece, 1 showing XX and 1 showing XY chromosomes. I'm not a
scientist nor do I have any understanding of what's involved! I do have a vague understanding that
it will be with an electron microscope, the slides would be of human dyed dead cells since, i
believe, when you show dna/chromosomes you have to break them apart or something. So I am
specifically looking to find someone with the expertise to assist me in understanding what the
process takes, what i'd exactly be looking at with the final video, and then to do the actual
preparing of slides and filming. If all possible, I'd also love to be in attendance to see the
process so I could learn and talk about the final piece with much more understanding. I know there
might be stock footage out there to use but i'd prefer first to see if I can be apart of actually
doing it as it helps me learn and have a much better understand and that is important to me as an
artist.

Can anyone help me in this endeavor? Or point me to someone that can? The work will be showing at
UW-Parkside in Wisconsin in November and will be comprised of photographic portraits of people,
several video pieces and an installation piece. I can pay for any supplies but this is all
personally funded so it would have to be reasonable. Thank you in advance for any assistance or help
with this matter.

Thanks, Lois Bielefeld

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Hi Rosemary,

We had Perspex canopies made up for our SEMs, Auger system and SIMS
machines to protect from direct sprinkler spray.

Regards, Barry

Barry Lamb
Director
Mead Testing Ltd
Registered address: 25 Mead Park,River Way, Harlow, Essex, CM20 2SE, UK
Company no. 4572600
barrylamb-at-meadtest.com
www.meadtest.com
tel: + 44 (0) 1279 635864
fax: +44 (0) 1279 635874

-----Original Message-----
X-from: protrain-at-emcourses.com [mailto:protrain-at-emcourses.com]
Sent: 27 June 2014 13:21
To: barrylamb-at-meadtest.com

Hi Rosemary

Out of interest I have dealt with water cascading over microscopes, yes
it
has happened twice in my 49 electron microscopy years. As a service
technician the main problem is not so much the water, but the debris
that
the water beings down onto the electrical circuits. Both of the
instruments
in question were running on vacuum only and managed to keep going
overnight
until security found the water running through the particular
laboratories.
One was a tap exploding, shooting a fountain of water into the EM room,
the
other the result of putting out a fire three levels above.

A third situation occurred when a water pipe came off one instrument in
a
basement EM unit, flooding the complete area to about 6 inches. Everyone
of
the instruments kept going, but as I arrived a technician with rubber
boots
and a long wooden pole was taking care of switching the most endangered
instruments off. However this was in the 1980s when most EM Units were
underground; the one I mentioned was also used as a Tornado shelter!

EM Units and water, great fun!

Steve

Steve Chapman FRMS
Protrain for Consultancy and Courses in Electron Microscopy
Mobile +44 (0) 7711 606967 Fax +44 (0)1280 814007
www.emcourses.com

-----Original Message-----
X-from: Rosemary.White-at-csiro.au [mailto:Rosemary.White-at-csiro.au]
Sent: 26 June 2014 11:09
To: protrain-at-emcourses.com

Hi Everyone,
Purdue University has an opening for an electron microscopy staff scientist in the Birck Nanotechnology Center.

Details can be found on the openings web site http://purdue.taleo.net/careersection/wl/jobdetail.ftl. The job number is 1401687

I can answer any general questions. If interested please apply immediately as the review process is about to begin.

Regards

Chris

Christopher J. Gilpin Ph.D.
Director, Life Science Microscopy Facility
Interim Director, Biological Sciences Electron Microscope Facility
Purdue University
Whistler Hall of Agriculture Research, Room S052
170 S. University St
West Lafayette, IN 47907
765-494-7750
gilpin-at-purdue.edu
http://www3.ag.purdue.edu/facilities/microscopy/Pages/default.aspx
https://www.bio.purdue.edu/molecular_biosciences/microscope_facility.html

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My question is is:
“How many people have experienced sparkler systems go off in their microscope rooms?”
Anyone want to jump in a relate experiences?

____
John Mansfield PhD Cphys MInstP
*****Note New Business Mailing Address, all other information remains the same*****
Microscopy Society of America - President-Elect
Associate Director
North Campus Electron Microbeam Analysis Laboratory
Building 22, Room G010, University of Michigan
2800 Plymouth Road, Ann Arbor MI 48109-2800 USA
Phone: (734) 936-3352 FAX (734) 763-2282
Cell. Phone: (734) 834-3913
Email: jfmjfm-at-umich.edu
URL: http://emalwww.engin.umich.edu/
iMessage: thejfmjfm
Skype: thejfmjfm

Home address:
4304 Spring Lake Boulevard
Ann Arbor MI 48108-9657
Phone (734) 994-3096

On Jun 27, 2014, at 9:17 AM, barrylamb-at-meadtest.com wrote:

}
}
}
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} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
} To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
} On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} ----------------------------------------------------------------------------
}
} Hi Rosemary,
}
} We had Perspex canopies made up for our SEMs, Auger system and SIMS
} machines to protect from direct sprinkler spray.
}
} Regards, Barry
}
} Barry Lamb
} Director
} Mead Testing Ltd
} Registered address: 25 Mead Park,River Way, Harlow, Essex, CM20 2SE, UK
} Company no. 4572600
} barrylamb-at-meadtest.com
} www.meadtest.com
} tel: + 44 (0) 1279 635864
} fax: +44 (0) 1279 635874
}
}
} -----Original Message-----
} X-from: protrain-at-emcourses.com [mailto:protrain-at-emcourses.com]
} Sent: 27 June 2014 13:21
} To: barrylamb-at-meadtest.com
} Subject: [Microscopy] RE: Sprinklers in instrument rooms
}
}
}
}
} ------------------------------------------------------------------------
} ----
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of
} America
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} http://www.microscopy.com/MicroscopyListserver
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} ------------------------------------------------------------------------
} ----
}
} Hi Rosemary
}
} Out of interest I have dealt with water cascading over microscopes, yes
} it
} has happened twice in my 49 electron microscopy years. As a service
} technician the main problem is not so much the water, but the debris
} that
} the water beings down onto the electrical circuits. Both of the
} instruments
} in question were running on vacuum only and managed to keep going
} overnight
} until security found the water running through the particular
} laboratories.
} One was a tap exploding, shooting a fountain of water into the EM room,
} the
} other the result of putting out a fire three levels above.
}
} A third situation occurred when a water pipe came off one instrument in
} a
} basement EM unit, flooding the complete area to about 6 inches. Everyone
} of
} the instruments kept going, but as I arrived a technician with rubber
} boots
} and a long wooden pole was taking care of switching the most endangered
} instruments off. However this was in the 1980s when most EM Units were
} underground; the one I mentioned was also used as a Tornado shelter!
}
} EM Units and water, great fun!
}
} Steve
}
} Steve Chapman FRMS
} Protrain for Consultancy and Courses in Electron Microscopy
} Mobile +44 (0) 7711 606967 Fax +44 (0)1280 814007
} www.emcourses.com
}
} -----Original Message-----
} X-from: Rosemary.White-at-csiro.au [mailto:Rosemary.White-at-csiro.au]
} Sent: 26 June 2014 11:09
} To: protrain-at-emcourses.com
} Subject: [Microscopy] Sprinklers in instrument rooms
}
} Hi all,
}
} I know this has been discussed before, just wondered if there were any
} new
} thoughts. We're moving twice, first into a refurbished basement, second
} time
} into a new building. The basement has a sprinkler system in case of
} fire,
} unlike our current building which just has alarms (it's a single storey
} brick building). I imagine the new building will have some sort of
} sprinkler
} system too.
}
} What precautions, if any, do people take to protect against the unlikely
} showering of your confocal or EM?
} Are there any relatively simple alternatives? (One alternative is to
} have
} fire-doors and walls for each room but that is prohibitive.)
}
} thanks much,
} Rosemary
}
} Dr Rosemary White
} CSIRO Plant Industry (for 4 more days only) GPO Box 1600 Canberra, ACT
} 2601
}
} T 02 6246 5475
} F 02 6246 5334
} E rosemary.white-at-csiro.au
}
}
}
} ==============================Original
} Headers==============================
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} 26 Jun 2014 20:06:24 +1000 8, 42 -- From: {Rosemary.White-at-csiro.au} 8,
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} instrument
} rooms 8, 42 -- Thread-Topic: Sprinklers in instrument rooms 8, 42 --
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} 28, 25 -- From barrylamb-at-meadtest.com Fri Jun 27 08:09:19 2014
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} 28, 25 -- Subject: RE: [Microscopy] RE: Sprinklers in instrument rooms
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Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

As hinted at from my post on the confocal listserver, I worked in an E. M. lab years ago in a hospital with a sprinkler head that developed a drip. fortunately, the head wasn't over the microscope, and was swapped out. In my new lab, the engineers and construction crew put a sprinkler head directly above my TEM column, which I made them move to the left wall of my microscope suite.

Ed

Edward Haller, Lab Manager
University of South Florida
Department of Integrative Biology
Electron Microscopy Core
SCA 110
4202 East Fowler Avenue
Tampa, FL 33620
(813)974-2676
ehaller-at-usf.edu
Office: ISA 1046
http://biology.usf.edu/ib/research/facilities/

________________________________________
X-from: jfmjfm-at-umich.edu [jfmjfm-at-umich.edu]
Sent: Friday, June 27, 2014 12:01 PM
To: Haller, Edward

My question is is:
“How many people have experienced sparkler systems go off in their microscope rooms?”
Anyone want to jump in a relate experiences?

____
John Mansfield PhD Cphys MInstP
*****Note New Business Mailing Address, all other information remains the same*****
Microscopy Society of America - President-Elect
Associate Director
North Campus Electron Microbeam Analysis Laboratory
Building 22, Room G010, University of Michigan
2800 Plymouth Road, Ann Arbor MI 48109-2800 USA
Phone: (734) 936-3352 FAX (734) 763-2282
Cell. Phone: (734) 834-3913
Email: jfmjfm-at-umich.edu
URL: http://emalwww.engin.umich.edu/
iMessage: thejfmjfm
Skype: thejfmjfm

Home address:
4304 Spring Lake Boulevard
Ann Arbor MI 48108-9657
Phone (734) 994-3096

On Jun 27, 2014, at 9:17 AM, barrylamb-at-meadtest.com wrote:

}
}
}
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
} To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
} On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} ----------------------------------------------------------------------------
}
} Hi Rosemary,
}
} We had Perspex canopies made up for our SEMs, Auger system and SIMS
} machines to protect from direct sprinkler spray.
}
} Regards, Barry
}
} Barry Lamb
} Director
} Mead Testing Ltd
} Registered address: 25 Mead Park,River Way, Harlow, Essex, CM20 2SE, UK
} Company no. 4572600
} barrylamb-at-meadtest.com
} www.meadtest.com
} tel: + 44 (0) 1279 635864
} fax: +44 (0) 1279 635874
}
}
} -----Original Message-----
} X-from: protrain-at-emcourses.com [mailto:protrain-at-emcourses.com]
} Sent: 27 June 2014 13:21
} To: barrylamb-at-meadtest.com
} Subject: [Microscopy] RE: Sprinklers in instrument rooms
}
}
}
}
} ------------------------------------------------------------------------
} ----
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of
} America
} To Subscribe/Unsubscribe --
} http://www.microscopy.com/MicroscopyListserver
} On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} ------------------------------------------------------------------------
} ----
}
} Hi Rosemary
}
} Out of interest I have dealt with water cascading over microscopes, yes
} it
} has happened twice in my 49 electron microscopy years. As a service
} technician the main problem is not so much the water, but the debris
} that
} the water beings down onto the electrical circuits. Both of the
} instruments
} in question were running on vacuum only and managed to keep going
} overnight
} until security found the water running through the particular
} laboratories.
} One was a tap exploding, shooting a fountain of water into the EM room,
} the
} other the result of putting out a fire three levels above.
}
} A third situation occurred when a water pipe came off one instrument in
} a
} basement EM unit, flooding the complete area to about 6 inches. Everyone
} of
} the instruments kept going, but as I arrived a technician with rubber
} boots
} and a long wooden pole was taking care of switching the most endangered
} instruments off. However this was in the 1980s when most EM Units were
} underground; the one I mentioned was also used as a Tornado shelter!
}
} EM Units and water, great fun!
}
} Steve
}
} Steve Chapman FRMS
} Protrain for Consultancy and Courses in Electron Microscopy
} Mobile +44 (0) 7711 606967 Fax +44 (0)1280 814007
} www.emcourses.com
}
} -----Original Message-----
} X-from: Rosemary.White-at-csiro.au [mailto:Rosemary.White-at-csiro.au]
} Sent: 26 June 2014 11:09
} To: protrain-at-emcourses.com
} Subject: [Microscopy] Sprinklers in instrument rooms
}
} Hi all,
}
} I know this has been discussed before, just wondered if there were any
} new
} thoughts. We're moving twice, first into a refurbished basement, second
} time
} into a new building. The basement has a sprinkler system in case of
} fire,
} unlike our current building which just has alarms (it's a single storey
} brick building). I imagine the new building will have some sort of
} sprinkler
} system too.
}
} What precautions, if any, do people take to protect against the unlikely
} showering of your confocal or EM?
} Are there any relatively simple alternatives? (One alternative is to
} have
} fire-doors and walls for each room but that is prohibitive.)
}
} thanks much,
} Rosemary
}
} Dr Rosemary White
} CSIRO Plant Industry (for 4 more days only) GPO Box 1600 Canberra, ACT
} 2601
}
} T 02 6246 5475
} F 02 6246 5334
} E rosemary.white-at-csiro.au
}
}
}
} ==============================Original
} Headers==============================
} 8, 42 -- From prvs=247c2aa8e=Rosemary.White-at-csiro.au Thu Jun 26 05:06:26
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} 26 Jun 2014 20:06:24 +1000 8, 42 -- From: {Rosemary.White-at-csiro.au} 8,
} 42 --
} To: {microscopy-at-microscopy.com} 8, 42 -- Subject: Sprinklers in
} instrument
} rooms 8, 42 -- Thread-Topic: Sprinklers in instrument rooms 8, 42 --
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} 17, 24 -- From protrain-at-emcourses.com Fri Jun 27 06:57:02 2014
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} 17, 24 -- To: {Rosemary.White-at-csiro.au}
} 17, 24 -- Cc: "'Microscopical Soc of America'"
} {microscopy-at-microscopy.com}
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} 17, 24 -- Subject: RE: [Microscopy] Sprinklers in instrument rooms
} 17, 24 -- Date: Fri, 27 Jun 2014 12:56:58 +0100
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15, 47 -- From ehaller-at-health.usf.edu Fri Jun 27 11:44:10 2014
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Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

At a previous job of mine, we had a sprinkler head spontaneously fail
catastrophically. The entire contents of the rooftop reservoir emptied into
the space, which had offices and labs (fortunately "just" test equipment).
Lots of water, and had a dispersion of fine black particulates in it.
Apparently this built up from reactions of water with the pipes over long
times. Very destructive.

-----Original Message-----
X-from: ehaller-at-health.usf.edu [mailto:ehaller-at-health.usf.edu]
Sent: Friday, June 27, 2014 12:55 PM
To: LES-at-ZSGENETICS.COM

As hinted at from my post on the confocal listserver, I worked in an E. M.
lab years ago in a hospital with a sprinkler head that developed a drip.
fortunately, the head wasn't over the microscope, and was swapped out. In my
new lab, the engineers and construction crew put a sprinkler head directly
above my TEM column, which I made them move to the left wall of my
microscope suite.

Ed

Edward Haller, Lab Manager
University of South Florida
Department of Integrative Biology
Electron Microscopy Core
SCA 110
4202 East Fowler Avenue
Tampa, FL 33620
(813)974-2676
ehaller-at-usf.edu
Office: ISA 1046
http://biology.usf.edu/ib/research/facilities/

________________________________________
X-from: jfmjfm-at-umich.edu [jfmjfm-at-umich.edu]
Sent: Friday, June 27, 2014 12:01 PM
To: Haller, Edward

My question is is:
"How many people have experienced sparkler systems go off in their
microscope rooms?"
Anyone want to jump in a relate experiences?

____
John Mansfield PhD Cphys MInstP
*****Note New Business Mailing Address, all other information remains the
same***** Microscopy Society of America - President-Elect Associate Director
North Campus Electron Microbeam Analysis Laboratory Building 22, Room G010,
University of Michigan
2800 Plymouth Road, Ann Arbor MI 48109-2800 USA
Phone: (734) 936-3352 FAX (734) 763-2282 Cell. Phone: (734) 834-3913
Email: jfmjfm-at-umich.edu
URL: http://emalwww.engin.umich.edu/
iMessage: thejfmjfm
Skype: thejfmjfm

Home address:
4304 Spring Lake Boulevard
Ann Arbor MI 48108-9657
Phone (734) 994-3096

On Jun 27, 2014, at 9:17 AM, barrylamb-at-meadtest.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
} ----------------------------------------------------------------------
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}
} Hi Rosemary,
}
} We had Perspex canopies made up for our SEMs, Auger system and SIMS
} machines to protect from direct sprinkler spray.
}
} Regards, Barry
}
} Barry Lamb
} Director
} Mead Testing Ltd
} Registered address: 25 Mead Park,River Way, Harlow, Essex, CM20 2SE,
} UK Company no. 4572600 barrylamb-at-meadtest.com www.meadtest.com
} tel: + 44 (0) 1279 635864
} fax: +44 (0) 1279 635874
}
}
} -----Original Message-----
} X-from: protrain-at-emcourses.com [mailto:protrain-at-emcourses.com]
} Sent: 27 June 2014 13:21
} To: barrylamb-at-meadtest.com
} Subject: [Microscopy] RE: Sprinklers in instrument rooms
}
}
}
}
} ----------------------------------------------------------------------
} --
} ----
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of
} America To Subscribe/Unsubscribe --
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} --
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}
} Hi Rosemary
}
} Out of interest I have dealt with water cascading over microscopes,
} yes it has happened twice in my 49 electron microscopy years. As a
} service technician the main problem is not so much the water, but the
} debris that the water beings down onto the electrical circuits. Both
} of the instruments in question were running on vacuum only and managed
} to keep going overnight until security found the water running through
} the particular laboratories.
} One was a tap exploding, shooting a fountain of water into the EM
} room, the other the result of putting out a fire three levels above.
}
} A third situation occurred when a water pipe came off one instrument
} in a basement EM unit, flooding the complete area to about 6 inches.
} Everyone of the instruments kept going, but as I arrived a technician
} with rubber boots and a long wooden pole was taking care of switching
} the most endangered instruments off. However this was in the 1980s
} when most EM Units were underground; the one I mentioned was also used
} as a Tornado shelter!
}
} EM Units and water, great fun!
}
} Steve
}
} Steve Chapman FRMS
} Protrain for Consultancy and Courses in Electron Microscopy
} Mobile +44 (0) 7711 606967 Fax +44 (0)1280 814007
} www.emcourses.com
}
} -----Original Message-----
} X-from: Rosemary.White-at-csiro.au [mailto:Rosemary.White-at-csiro.au]
} Sent: 26 June 2014 11:09
} To: protrain-at-emcourses.com
} Subject: [Microscopy] Sprinklers in instrument rooms
}
} Hi all,
}
} I know this has been discussed before, just wondered if there were any
} new thoughts. We're moving twice, first into a refurbished basement,
} second time into a new building. The basement has a sprinkler system
} in case of fire, unlike our current building which just has alarms
} (it's a single storey brick building). I imagine the new building will
} have some sort of sprinkler system too.
}
} What precautions, if any, do people take to protect against the
} unlikely showering of your confocal or EM?
} Are there any relatively simple alternatives? (One alternative is to
} have fire-doors and walls for each room but that is prohibitive.)
}
} thanks much,
} Rosemary
}
} Dr Rosemary White
} CSIRO Plant Industry (for 4 more days only) GPO Box 1600 Canberra, ACT
} 2601
}
} T 02 6246 5475
} F 02 6246 5334
} E rosemary.white-at-csiro.au
}
}
}
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} 28, 25 -- From barrylamb-at-meadtest.com Fri Jun 27 08:09:19 2014 28, 25
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-0500
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} 28, 25 -- Fri, 27 Jun 2014 14:09:17 +0100 (BST)
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15, 47 -- From ehaller-at-health.usf.edu Fri Jun 27 11:44:10 2014 15, 47 --
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Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

I just love the Apple autocorrect!
Sprinkler systems. I am willing to bet that NO-ONE has experienced an accidental release of water from a sprinkler system.
Flooding from other labs or burst pipes, yes, but sprinklers I think not.

____
John Mansfield PhD Cphys MInstP
*****Note New Business Mailing Address, all other information remains the same*****
Microscopy Society of America - President-Elect
Associate Director
North Campus Electron Microbeam Analysis Laboratory
Building 22, Room G010, University of Michigan
2800 Plymouth Road, Ann Arbor MI 48109-2800 USA
Phone: (734) 936-3352 FAX (734) 763-2282
Cell. Phone: (734) 834-3913
Email: jfmjfm-at-umich.edu
URL: http://emalwww.engin.umich.edu/
iMessage: thejfmjfm
Skype: thejfmjfm

Home address:
4304 Spring Lake Boulevard
Ann Arbor MI 48108-9657
Phone (734) 994-3096

On Jun 27, 2014, at 12:26 PM, Philip Oshel {oshel1pe-at-cmich.edu} wrote:

} John,
}
} A very relevant question, given the 4th of July is next week, but I don't think many people have "sparkler systems" in their microscopy labs.
} Would be pretty and exciting, though ...
}
} Phil
}
}
} } My question is is:
} } “How many people have experienced sparkler systems go off in their microscope rooms?”
} } Anyone want to jump in a relate experiences?
} }
} }
} } ____
} } John Mansfield PhD Cphys MInstP
} } *****Note New Business Mailing Address, all other information remains the same*****
} } Microscopy Society of America - President-Elect
} } Associate Director
} } North Campus Electron Microbeam Analysis Laboratory
} } Building 22, Room G010, University of Michigan
} } 2800 Plymouth Road, Ann Arbor MI 48109-2800 USA
} } Phone: (734) 936-3352 FAX (734) 763-2282
} } Cell. Phone: (734) 834-3913
} } Email: jfmjfm-at-umich.edu
} } URL: http://emalwww.engin.umich.edu/
} } iMessage: thejfmjfm
} } Skype: thejfmjfm
} }
} } Home address:
} } 4304 Spring Lake Boulevard
} } Ann Arbor MI 48108-9657
} } Phone (734) 994-3096
} }
} } On Jun 27, 2014, at 9:17 AM, barrylamb-at-meadtest.com wrote:
} }
} } }
} } }
} } }
} } } ----------------------------------------------------------------------------
} } } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
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} } }
} } } Hi Rosemary,
} } }
} } } We had Perspex canopies made up for our SEMs, Auger system and SIMS
} } } machines to protect from direct sprinkler spray.
} } }
} } } Regards, Barry
} } }
} } } Barry Lamb
} } } Director
} } } Mead Testing Ltd
} } } Registered address: 25 Mead Park,River Way, Harlow, Essex, CM20 2SE, UK
} } } Company no. 4572600
} } } barrylamb-at-meadtest.com
} } } www.meadtest.com
} } } tel: + 44 (0) 1279 635864
} } } fax: +44 (0) 1279 635874
} } }
} } }
} } } -----Original Message-----
} } } X-from: protrain-at-emcourses.com [mailto:protrain-at-emcourses.com]
} } } Sent: 27 June 2014 13:21
} } } To: barrylamb-at-meadtest.com
} } } Subject: [Microscopy] RE: Sprinklers in instrument rooms
} } }
} } }
} } }
} } }
} } } ------------------------------------------------------------------------
} } } ----
} } } The Microscopy ListServer -- CoSponsor: The Microscopy Society of
} } } America
} } } To Subscribe/Unsubscribe --
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} } } ----
} } }
} } } Hi Rosemary
} } }
} } } Out of interest I have dealt with water cascading over microscopes, yes
} } } it
} } } has happened twice in my 49 electron microscopy years. As a service
} } } technician the main problem is not so much the water, but the debris
} } } that
} } } the water beings down onto the electrical circuits. Both of the
} } } instruments
} } } in question were running on vacuum only and managed to keep going
} } } overnight
} } } until security found the water running through the particular
} } } laboratories.
} } } One was a tap exploding, shooting a fountain of water into the EM room,
} } } the
} } } other the result of putting out a fire three levels above.
} } }
} } } A third situation occurred when a water pipe came off one instrument in
} } } a
} } } basement EM unit, flooding the complete area to about 6 inches. Everyone
} } } of
} } } the instruments kept going, but as I arrived a technician with rubber
} } } boots
} } } and a long wooden pole was taking care of switching the most endangered
} } } instruments off. However this was in the 1980s when most EM Units were
} } } underground; the one I mentioned was also used as a Tornado shelter!
} } }
} } } EM Units and water, great fun!
} } }
} } } Steve
} } }
} } } Steve Chapman FRMS
} } } Protrain for Consultancy and Courses in Electron Microscopy
} } } Mobile +44 (0) 7711 606967 Fax +44 (0)1280 814007
} } } www.emcourses.com
} } }
} } } -----Original Message-----
} } } X-from: Rosemary.White-at-csiro.au [mailto:Rosemary.White-at-csiro.au]
} } } Sent: 26 June 2014 11:09
} } } To: protrain-at-emcourses.com
} } } Subject: [Microscopy] Sprinklers in instrument rooms
} } }
} } } Hi all,
} } }
} } } I know this has been discussed before, just wondered if there were any
} } } new
} } } thoughts. We're moving twice, first into a refurbished basement, second
} } } time
} } } into a new building. The basement has a sprinkler system in case of
} } } fire,
} } } unlike our current building which just has alarms (it's a single storey
} } } brick building). I imagine the new building will have some sort of
} } } sprinkler
} } } system too.
} } }
} } } What precautions, if any, do people take to protect against the unlikely
} } } showering of your confocal or EM?
} } } Are there any relatively simple alternatives? (One alternative is to
} } } have
} } } fire-doors and walls for each room but that is prohibitive.)
} } }
} } } thanks much,
} } } Rosemary
} } }
} } } Dr Rosemary White
} } } CSIRO Plant Industry (for 4 more days only) GPO Box 1600 Canberra, ACT
} } } 2601
} } }
} } } T 02 6246 5475
} } } F 02 6246 5334
} } } E rosemary.white-at-csiro.au
} } }
} } }
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} } } 28, 25 -- From barrylamb-at-meadtest.com Fri Jun 27 08:09:19 2014
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} } .
} }
}
} --
} Philip Oshel
} Microscopy Facility Supervisor
} Biology Department
} 024C Brooks Hall
} Central Michigan University
} Mt. Pleasant, MI 48859
} (989) 774-3576
}

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The previous link did not work for some

Try this link to the search page

http://purdue.taleo.net/careersection/wl/moresearch.ftl#

The job number is 1401687

Christopher J. Gilpin Ph.D.
Director, Life Science Microscopy Facility
Interim Director, Biological Sciences Electron Microscope Facility
Purdue University
Whistler Hall of Agriculture Research, Room S052
170 S. University St
West Lafayette, IN 47907
765-494-7750
gilpin-at-purdue.edu
http://www3.ag.purdue.edu/facilities/microscopy/Pages/default.aspx
https://www.bio.purdue.edu/molecular_biosciences/microscope_facility.html

==============================Original Headers==============================
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Hi there,

Do you have experience with HPF-FS of petals from mature flowers? If so
than I would appreciate your contacting me as I have some technical
questions to ask you. We are primarily interested in the epithelial layer
and this is a challenge since there is so little cytoplasm.

Thanks,
Debby

Debra Sherman, Founder & CSO
DS imaging LLC
Purdue Technology Center
1281 Win Hentschel Blvd
West Lafayette, IN 47906
E-mail: debby.sherman-at-dsimagingllc.com
www.dsimagingllc.com
Mobile: 765-418-8540

==============================Original Headers==============================
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8, 31 -- From: "Sherman, Debra" {dsherman-at-purdue.edu}
8, 31 -- To: "message to: MSA list" {microscopy-at-microscopy.com}
8, 31 -- Subject: HPF of flower petals
8, 31 -- Thread-Topic: HPF of flower petals
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From pharmacy-popular11-at-charter.com Fri Jun 27 19:34:38 2014
Return-Path: {pharmacy-popular11-at-charter.com}
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X-from: dan.fairweather-at-delphi.com ()

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Email: dan.fairweather-at-delphi.com
Name: Dan Fairweather

Organization: Delphi Automotive, Powertrain Div

Title-Subject: [Filtered] S3200N spare parts?

Message: It looks like we need a new needle valve for our Hitachi S3200N. We have one on order with
a promised delivery of late August. I was wondering if there is any one out there who might have
something on the shelf, or maybe a "mildly" used one.

The Hitachi part number is 49E-7391.

Thank you for your assistance,
Dan Fairweather
Senior Materials Development Engineer
Delphi Powertrain

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Email: scalliom-at-myumanitoba.ca
Name: Melanie Scallion

Organization: University of Manitoba

Title-Subject: [Filtered] SEM - Need help with which scope to use for insects

Message: I am an undergraduate student at the University of Manitoba working on a revision of a
genus of parasitic wasps. I have three new species that I would like to take images of in order to
see their sculpturing, and I’m trying to figure out which SEM I should use. The specimens are about
3mm in length, and because they are not mine, I need them to remain as is (they have to be returned
in the same condition in which I received them). The insects are point mounted, which means they are
glued to the end of a small, pointed piece of paper which then has a pin stuck through it. The glue
used likely dissolves in ethanol.

The two scopes we have here at the university are: Philips XL 30 and JEOL JSM-5900LV. I’m hoping
someone could provide me with a recommendation on which to use, and advise me on what I will need to
do if one of these scopes is suitable.

Any information would be of great help! Thank you,

Melanie Scallion

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Email: roger.ristau-at-uconn.edu
Name: Roger Ristau

Organization: University of Connecticut

Title-Subject: [Filtered] Re: Sprinklers in instrument rooms

Message: Here is a situation that I bet not many have encountered: We had a TEM which developed a
leak in lens cooling with the result that the INSIDE of the TEM filled with water. (I was not here
but was told the camera/viewing chamber looked like "a fish bowl.")

My predecessor, Larry McCurdy, courageously attacked the mess and restored the TEM to service! It
soldiered on for many years.

Point of story: Flooding the OUTSIDE of the TEM may actually be more harmful (electrical
shorts,etc.) than flooding the INSIDE!

Cheers
Roger

Institute of Materials Science
University of Connecticut

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Email: rashmi_mehata-at-yahoo.com
Name: Rashmi

Organization: DBT

Title-Subject: [Filtered] beam fluctuation in FEI T20

Message: Hello,

Variation in beam intensity is observed with LaB6. How ever the LaB6 filament is showing the
connectivity. What could be the possible problem.

Rashmi

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Melanie - I think you are going to have a tough time viewing these in an SEM in a way that leaves them in an untouched condition. An alternative would be to see if you can get access to a high quality stereomicroscope - ideally one with software that allows extended depth of focus (EDF) images to be taken. One problem with a conventional stereoscope is that you can see the top of the insect in focus but not the bottom or vice versa. With EDF software, one can grab a digital image of each focal plane and the software reconstructs an image that is in good focus for the entire depth of the specimen. We have a motorized Leica MTOF FA stereomicroscope in our core facility and clients working with insects love it. You can see some examples of images taken with and without EDF on this microscope at our website http://www.biotech.missouri.edu/mcc/Stereoscope_M205.html . Good luck. Tom

Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
phillipst-at-missouri.edu

http://www.biology.missouri.edu/faculty/phillips.html
http://www.biotech.missouri.edu/mcc/

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Email: scalliom-at-myumanitoba.ca
Name: Melanie Scallion

Organization: University of Manitoba

Title-Subject: [Filtered] SEM - Need help with which scope to use for insects

Message: I am an undergraduate student at the University of Manitoba working on a revision of a genus of parasitic wasps. I have three new species that I would like to take images of in order to see their sculpturing, and I'm trying to figure out which SEM I should use. The specimens are about 3mm in length, and because they are not mine, I need them to remain as is (they have to be returned in the same condition in which I received them). The insects are point mounted, which means they are glued to the end of a small, pointed piece of paper which then has a pin stuck through it. The glue used likely dissolves in ethanol.

The two scopes we have here at the university are: Philips XL 30 and JEOL JSM-5900LV. I'm hoping someone could provide me with a recommendation on which to use, and advise me on what I will need to do if one of these scopes is suitable.

Any information would be of great help! Thank you,

Melanie Scallion

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Hi Melanie,
Tom's suggestion to use EDF is a good one. Those motorized Leica
systems are excellent and pricey. Good manual alternatives are
available at much lower cost. A good DSLR kit on a stereo with Helicon
focus or other free or inexpensive software packages do a good job.
Just ensure the software has a stereo microscope plugin to account for
the lateral image shift as the stereo is focused.
Jim

--
JIM SCHULTE

Micro Source Imaging
612 770-3885
www.ms-imaging.com
Sales and service of Canon, Motic and Diagnostic Instruments cameras
Motic, Accu-scope and Unitron microscopes

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From drugs_canadian3-at-irtel.ru Sat Jun 28 18:02:56 2014
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FYI a summary of responses. Thanks much to all, very useful.

1. Alternatives are available, but expensive. These include:
- firewalls around each room lacking sprinklers, and firedoor entryway -
rather expensive. The walls and door have to be able to resist fire for 1
hour.
- gas systems - also expensive, and in Australia these are apparently only
allowed in non-occupied rooms (I am told, not going to fight it, save
fight for other stuff...).

So

2. Accept the inevitability of sprinklers, and insist on the following:
- sprinkler head(s) as far from instrument as possible, esp. from main
power supplies
- sprinkler heads enclosed in wire cages to prevent accidental knocking
when moving large items in the room

3. Realise that floods from pipes/sinks/rain events are much more likely
than sprinkler activation, so:
- install drip pans above the sprinklers, either below the ceiling or
within it, to deflect potential floods from above away from the instrument
- raise important components at least several cm above floor level
- have a system that only fills the sprinkler pipes when the sprinkler(s)
are activated to avoid slow leaks (drips)

4. Also realise that sprinkler pipes may be sources of conductive
interference, so:
- ask that pipes are empty unless sprinkler(s) activated
- insert a rubber or other non-metallic connection between the main
sprinkler supply system and the sprinklers in instrument rooms

and also, check your insurance policy!

Like others, the only floods I've experienced were from other sources - in
a previous institution on one torrential weekend several staff came in to
check for leaks and ended up doing a bucket brigade from the EM unit in
the basement. All fine, except for the piles of paperwork around desks...

And Roger Ristau's email reminded me that about 15 years ago the in lens
cooling system in our JEOL 6400 developed a leak and flooded the lens,
detected by water dripping down into the chamber - messy! The old girl is
still chugging along, though not for much longer - the old electronic
boards are cracking and won't survive the move, so turning off for good in
October. :(

cheers,
Rosemary

Dr Rosemary White
CSIRO Plant Industry (for 1 more day)
GPO Box 1600
Canberra, ACT 2601
Australia

T 61 2 6246 5475
F 61 2 6246 5334
E rosemary.white-at-csiro.au

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I assume not too many rooms have been flooded from accidental releases from sparkler systems, but I am aware of some accidental releases from sprinkler systems.

I had a project come through years ago where a consulting engineer was looking at the metal fuse in a sprinkler element. It had released a flooded a store. His question was whether its release was prompted by a thermal event or a mechanical event. I suppose both would show some signs of ductile rupture and he did not have an exemplar for us to examine from the two possibilities.

On the flood story side, we are waiting for the plumbers to resolved a problem in the bathroom upstairs. Someone saw fit to replumb with low-flow valves. However, the replumbing has failed on at least two occasions leading to lots of "clean" water coming through the floor into our lab below. I don't know what the savings in water would have been but the initial costs of replumbing, but the costs of clean-up and the necessary replacement of ceiling tiles has certainly lengthened out the pay-back period. I am really leery of "upgrades".

Warren

-----Original Message-----
X-from: jfmjfm-at-umich.edu [mailto:jfmjfm-at-umich.edu]
Sent: Friday, June 27, 2014 2:47 PM
To: Straszheim, Warren E [BIOTC]

I just love the Apple autocorrect!
Sprinkler systems. I am willing to bet that NO-ONE has experienced an accidental release of water from a sprinkler system.
Flooding from other labs or burst pipes, yes, but sprinklers I think not.

____
John Mansfield PhD Cphys MInstP
*****Note New Business Mailing Address, all other information remains the same*****
Microscopy Society of America - President-Elect
Associate Director
North Campus Electron Microbeam Analysis Laboratory
Building 22, Room G010, University of Michigan
2800 Plymouth Road, Ann Arbor MI 48109-2800 USA
Phone: (734) 936-3352 FAX (734) 763-2282
Cell. Phone: (734) 834-3913
Email: jfmjfm-at-umich.edu
URL: http://emalwww.engin.umich.edu/
iMessage: thejfmjfm
Skype: thejfmjfm

Home address:
4304 Spring Lake Boulevard
Ann Arbor MI 48108-9657
Phone (734) 994-3096

On Jun 27, 2014, at 12:26 PM, Philip Oshel {oshel1pe-at-cmich.edu} wrote:

} John,
}
} A very relevant question, given the 4th of July is next week, but I don't think many people have "sparkler systems" in their microscopy labs.
} Would be pretty and exciting, though ...
}
} Phil
}
}
} } My question is is:
} } "How many people have experienced sparkler systems go off in their microscope rooms?"
} } Anyone want to jump in a relate experiences?
} }
} }
} } ____
} } John Mansfield PhD Cphys MInstP
} } *****Note New Business Mailing Address, all other information remains the same*****
} } Microscopy Society of America - President-Elect
} } Associate Director
} } North Campus Electron Microbeam Analysis Laboratory
} } Building 22, Room G010, University of Michigan
} } 2800 Plymouth Road, Ann Arbor MI 48109-2800 USA
} } Phone: (734) 936-3352 FAX (734) 763-2282
} } Cell. Phone: (734) 834-3913
} } Email: jfmjfm-at-umich.edu
} } URL: http://emalwww.engin.umich.edu/
} } iMessage: thejfmjfm
} } Skype: thejfmjfm
} }
} } Home address:
} } 4304 Spring Lake Boulevard
} } Ann Arbor MI 48108-9657
} } Phone (734) 994-3096
} }
} } On Jun 27, 2014, at 9:17 AM, barrylamb-at-meadtest.com wrote:
} }
} } }
} } }
} } }
} } } ----------------------------------------------------------------------------
} } } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
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} } } ----------------------------------------------------------------------------
} } }
} } } Hi Rosemary,
} } }
} } } We had Perspex canopies made up for our SEMs, Auger system and SIMS
} } } machines to protect from direct sprinkler spray.
} } }
} } } Regards, Barry
} } }
} } } Barry Lamb
} } } Director
} } } Mead Testing Ltd
} } } Registered address: 25 Mead Park,River Way, Harlow, Essex, CM20 2SE, UK
} } } Company no. 4572600
} } } barrylamb-at-meadtest.com
} } } www.meadtest.com
} } } tel: + 44 (0) 1279 635864
} } } fax: +44 (0) 1279 635874
} } }
} } }
} } } -----Original Message-----
} } } X-from: protrain-at-emcourses.com [mailto:protrain-at-emcourses.com]
} } } Sent: 27 June 2014 13:21
} } } To: barrylamb-at-meadtest.com
} } } Subject: [Microscopy] RE: Sprinklers in instrument rooms
} } }
} } }
} } }
} } }
} } } ------------------------------------------------------------------------
} } } ----
} } } The Microscopy ListServer -- CoSponsor: The Microscopy Society of
} } } America
} } } To Subscribe/Unsubscribe --
} } } http://www.microscopy.com/MicroscopyListserver
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} } } ------------------------------------------------------------------------
} } } ----
} } }
} } } Hi Rosemary
} } }
} } } Out of interest I have dealt with water cascading over microscopes, yes
} } } it
} } } has happened twice in my 49 electron microscopy years. As a service
} } } technician the main problem is not so much the water, but the debris
} } } that
} } } the water beings down onto the electrical circuits. Both of the
} } } instruments
} } } in question were running on vacuum only and managed to keep going
} } } overnight
} } } until security found the water running through the particular
} } } laboratories.
} } } One was a tap exploding, shooting a fountain of water into the EM room,
} } } the
} } } other the result of putting out a fire three levels above.
} } }
} } } A third situation occurred when a water pipe came off one instrument in
} } } a
} } } basement EM unit, flooding the complete area to about 6 inches. Everyone
} } } of
} } } the instruments kept going, but as I arrived a technician with rubber
} } } boots
} } } and a long wooden pole was taking care of switching the most endangered
} } } instruments off. However this was in the 1980s when most EM Units were
} } } underground; the one I mentioned was also used as a Tornado shelter!
} } }
} } } EM Units and water, great fun!
} } }
} } } Steve
} } }
} } } Steve Chapman FRMS
} } } Protrain for Consultancy and Courses in Electron Microscopy
} } } Mobile +44 (0) 7711 606967 Fax +44 (0)1280 814007
} } } www.emcourses.com
} } }
} } } -----Original Message-----
} } } X-from: Rosemary.White-at-csiro.au [mailto:Rosemary.White-at-csiro.au]
} } } Sent: 26 June 2014 11:09
} } } To: protrain-at-emcourses.com
} } } Subject: [Microscopy] Sprinklers in instrument rooms
} } }
} } } Hi all,
} } }
} } } I know this has been discussed before, just wondered if there were any
} } } new
} } } thoughts. We're moving twice, first into a refurbished basement, second
} } } time
} } } into a new building. The basement has a sprinkler system in case of
} } } fire,
} } } unlike our current building which just has alarms (it's a single storey
} } } brick building). I imagine the new building will have some sort of
} } } sprinkler
} } } system too.
} } }
} } } What precautions, if any, do people take to protect against the unlikely
} } } showering of your confocal or EM?
} } } Are there any relatively simple alternatives? (One alternative is to
} } } have
} } } fire-doors and walls for each room but that is prohibitive.)
} } }
} } } thanks much,
} } } Rosemary
} } }
} } } Dr Rosemary White
} } } CSIRO Plant Industry (for 4 more days only) GPO Box 1600 Canberra, ACT
} } } 2601
} } }
} } } T 02 6246 5475
} } } F 02 6246 5334
} } } E rosemary.white-at-csiro.au
} } }
} } }
} } }
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} } } 17, 24 -- From protrain-at-emcourses.com Fri Jun 27 06:57:02 2014
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} } } ==============================Original Headers==============================
} } } 28, 25 -- From barrylamb-at-meadtest.com Fri Jun 27 08:09:19 2014
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} } 8, 37 -- Subject: Re: [Microscopy] Sprinklers in instrument rooms
} } 8, 37 -- From: John Mansfield {jfmjfm-at-umich.edu}
} } 8, 37 -- In-Reply-To: {201406271317.s5RDH02h020872-at-ns.microscopy.com}
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} } .
} }
}
} --
} Philip Oshel
} Microscopy Facility Supervisor
} Biology Department
} 024C Brooks Hall
} Central Michigan University
} Mt. Pleasant, MI 48859
} (989) 774-3576
}

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Melanie:

I agree that the light microscope and EDF software would
produce some great images. I have produced some fun images
of bees, flies, and mosquitos with a stereo microscope and
free edf software called Combined ZM. The natural color of
the light microscope images adds to the image compared to
gray-scale SEM images for low-mag work.

For SEM, the JEOL 5900LV will be your best bet for
examination in an undisturbed state. In the LV mode, you can
observe the wasps without coating and should be able to get
some good images of general physical characteristics. You
will not be able to get real high magnification images since
you will be limited to backscattered imaging You would have
to sputter coat the samples to image secondary electrons in
the 5900LV or the Philips XL 30 for higher magnifications.
If you can sputter coat a specimen, you should be able to
get great images of the fine details with the Philips.

Good luck.

On 6/28/2014 7:35 AM,
microscopylistserver-noreply-at-microscopy.com wrote:
} Email:scalliom-at-myumanitoba.ca
} Name: Melanie Scallion
}
} Organization: University of Manitoba
}
} Title-Subject: [Filtered] SEM - Need help with which scope to use for insects
}
} Message: I am an undergraduate student at the University of Manitoba working on a revision of a
} genus of parasitic wasps. I have three new species that I would like to take images of in order to
} see their sculpturing, and I’m trying to figure out which SEM I should use. The specimens are about
} 3mm in length, and because they are not mine, I need them to remain as is (they have to be returned
} in the same condition in which I received them). The insects are point mounted, which means they are
} glued to the end of a small, pointed piece of paper which then has a pin stuck through it. The glue
} used likely dissolves in ethanol.
}
} The two scopes we have here at the university are: Philips XL 30 and JEOL JSM-5900LV. I’m hoping
} someone could provide me with a recommendation on which to use, and advise me on what I will need to
} do if one of these scopes is suitable.
}
} Any information would be of great help! Thank you,
}
} Melanie Scallion

--
________
Larry D. Hanke, P.E.
Principal Engineer
Materials Evaluation and Engineering, Inc.
(763) 449-8870

==============================Original Headers==============================
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From drugstore_discounted6-at-expertiserealty.com Sun Jun 29 20:13:39 2014
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Hi all,

As mentioned, we're eventually moving into a new building, and if
possible, we'll incorporate all the best building recommendations. I
remember when all TEM and SEM rooms had floating floors to eliminate or at
least substantially reduce vibrational noise. Is this still the best
option? It's rather expensive and if not really needed we could divert the
saved funds elsewhere. We are planning to have a plant room in which all
the noisy components - pumps and fans, mainly, are isolated from the main
instruments.

We have a VP-SEM and have a FESEM on the to-buy list for 2-3 years' time.
No TEM any more. We'll be on the ground floor and it's a fairly
vibration-free site - no main roads nearby, etc. Just need to make sure
the threshing and grinding equipment isn't installed next door...

thanks much,
Rosemary

Dr Rosemary White
CSIRO Plant Industry (last day)
GPO Box 1600
Canberra, ACT 2601

T 02 6246 5475
F 02 6246 5334
E rosemary.white-at-csiro.au

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Dear Roger,
dear all,

I would like to inform that the same
(micro-leak in the cooling line at the junction plastic hose to
copper-fitting of the projective lens of my ZEISS EM109) happened
in our EM-Lab (it was 18th April 2000 after nearly 20 years of operation).

It took some hours to get rid of all the water within the viewing
chamber and the 6x9 camera beneath the viewing chamber.

Needed a new projective lens, which is in function until now...
Not remembering what the Cost for repair were.
So, I think it will be time to either have an eye on it or to put it
into mothballs.....(;-()

Have a great and accident-free week,
regards,
Wolfgang

Wolfgang Muss
SALZBURG-AUSTRIA

} -----Ursprüngliche Nachricht-----
} Von: microscopylistserver-noreply-at-microscopy.com
} [mailto:microscopylistserver-noreply-at-microscopy.com]
} Gesendet: Samstag, 28. Juni 2014 14:42
} An: Muß Wolfgang
} Betreff: [Microscopy] Re: Sprinklers in instrument rooms
}
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} Email: roger.ristau-at-uconn.edu Name: Roger Ristau
} Organization: University of Connecticut
} Title-Subject: Re: Sprinklers in instrument
} rooms
} Message:
}
} Here is a situation that I bet not many have encountered:
} We had a TEM which developed a leak in lens cooling with the result
} that the INSIDE of the TEM filled with water. (I was not here
} but was told the camera/viewing chamber looked like "a fish bowl.")
}
} My predecessor, Larry McCurdy, courageously attacked the mess and
} restored the TEM to service! It soldiered on for many years.
} Point of story: Flooding the OUTSIDE of the TEM may actually be more
} harmful (electrical shorts,etc.) than flooding the INSIDE!
}
} Cheers
} Roger
}
} Institute of Materials Science
} University of Connecticut
}
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Dear Rosemary,

We moved the department wholesale into a new building last August.
During the design stage I was heavily involved in benchmarking good
laboratories with the structural engineers (Ramboll, www.ranboll.co.uk)
and our vibration/noise consultant, Michael Gendreau of Colin Gordons &
Associates (www.colingordon.com).

Michael was instrumental in our decision to do without isolation plinths
when he demonstrated that, in the low frequency regime, i.e. below the
resonant frequency of the plinth, the amplitude of the vibration was
worse than the outside structure, i.e. it was an amplifier of noise.
Above the resonant frequency, the plinth attenuated it, as expected
(referring to a damped harmonic oscillator). The Q-factor of each plinth
was determined by the damping material underneath the plinth. In some
cases the isolation block was moving 100 times faster (at resonance)
than the surrounding building (in one lab we tested).

The money was spent on making the single raft slab thicker - we doubled
it from the early design thickness of 1 metre to 2 metres thick and that
is what we have now (the main department building is 0.5m thick).
Further, we have separate room-within-a-room build with each microscope
having a cross-laminate timber walls and ceiling that are 20cm thick.
The rooms were finsihed with acoustic panels on the ceiling and 2 walls
to reduce noise. Room cooling was with a chilled ceiling (95% cooling)
with two plenums for a little cool fresh air. For a view of the room,
see here:

http://www-hrem.msm.cam.ac.uk/facilities/cm30/index.shtml

We have found that this works really well for us. For example, we were
getting information out to 0.9 Angstrom with an FEI Titan TEM in the old
building, but now, in the new, heavy, foundation we get information down
to 0.6 to 0.7 Angstrom.

In summary, the foundation of the building is as big and heavy as we
could make it and we avoided floating tables/isolation plinths. Michael
acknowledged that this was what they were recommending ten-to-fifteen
years ago, but have decided against it since when evidence accumulated
that they only work in certain circumstances.

I hope this helps.

Jon

} Hi all,
}
} As mentioned, we're eventually moving into a new building, and if
} possible, we'll incorporate all the best building recommendations. I
} remember when all TEM and SEM rooms had floating floors to eliminate or
} at
} least substantially reduce vibrational noise. Is this still the best
} option? It's rather expensive and if not really needed we could divert
} the
} saved funds elsewhere. We are planning to have a plant room in which
} all
} the noisy components - pumps and fans, mainly, are isolated from the
} main
} instruments.
}
} We have a VP-SEM and have a FESEM on the to-buy list for 2-3 years'
} time.
} No TEM any more. We'll be on the ground floor and it's a fairly
} vibration-free site - no main roads nearby, etc. Just need to make sure
} the threshing and grinding equipment isn't installed next door...
}
} thanks much,
} Rosemary
}
} Dr Rosemary White
} CSIRO Plant Industry (last day)
} GPO Box 1600
} Canberra, ACT 2601
}
} T 02 6246 5475
} F 02 6246 5334
} E rosemary.white-at-csiro.au

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11, 31 -- From jsb43-at-cam.ac.uk Mon Jun 30 06:49:31 2014
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Hi Rashmi

There are many reasons why a tip may become unstable but one of the most
common is a build up of material inside the cathode aperture.

Good luck

Steve

Steve Chapman FRMS
Protrain for Consultancy and Courses in Electron Microscopy
Mobile +44 (0) 7711 606967 Fax +44 (0)1280 814007
www.emcourses.com

Email: rashmi_mehata-at-yahoo.com
Name: Rashmi

Organization: DBT

Title-Subject: [Filtered] beam fluctuation in FEI T20

Message: Hello,

Variation in beam intensity is observed with LaB6. How ever the LaB6
filament is showing the connectivity. What could be the possible problem.

Rashmi

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Hi, Melanie,

I would recommend that you contact Sam Droeg. He works for the U. S. Geological Survey Department, and has developed a technique and hardware system for macrophotography of insects. He has posted remarkable insect photos on the web, and can talk you through your macrophotography need with a digital camera and macro lens. His contact information is: sdroege-at-usgs.gov 301 497 5840

You can see some of his work at:

USGSBIML site:

http://www.flickr.com/photos/usgsbiml/ USGS macrophotograph site.

Best pictures at USGSBIML (eye candy set):

http://www.flickr.com/photos/usgsbiml/sets/72157630468783226/ Macrophotographs of insects.

Specimens taken in Hand Sanitizer:

http://www.flickr.com/photos/usgsbiml/sets/72157631861396270/ Macrophotographs of specimens taken in cuvettes filled with hand sanitizer.

Perhaps the best technical forum on macrophotography:

http://www.photomacrography.net/ Online blog of photographers who do macrophotography.

Youtube on basic USGSBIML set up:

http://www.youtube.com/watch?v=S-_yvIsucOY Video on setting up a camera for macrophotography.

USGSBIML Photoshopping technique: Note that he now has added using the color burn brush at 20% opacity to clean up the halos that bleed into the black background from "hot" color sections of the picture.

http://www.youtube.com/watch?v=Bdmx_8zqvN4 Video of how to use Adobe Bridge and Photoshop to clean up photos after image stacking.

PDF of basic USGSBIML photography set up:

ftp://ftpext.usgs.gov/pub/er/md/laurel/Droege/How%20to%20Take%20MacroPhotographs%20of%20Insects%20BIML%20Lab2.pdf PDF file with description of camera and lens setup for macrophotography.

Google Hangout demonstration of techniques
https://plus.google.com/events/c5569losvskrv2nu606ltof8odo
or
http://www.youtube.com/watch?v=4c15neFttoU 47 minute interview showing the entire procedure of photography, with camera setup.

He uses a 3-D stitching software program called Zerene Stacker (although you could use Photoshop or other programs that handle 3-D images, if you are familiar with the software):

Zerene Stacker â€” Purchasing Licenses
http://zerenesystems.com/cms/stacker/docs/purchasing

Zerene Stacker has several advantages over other commonly used stacking programs:
â€¢ Highest quality output images, especially in difficult cases
o Accurate and robust alignment and interpolation
o Advanced stacking algorithms
ï‚§ Clean handling of hairs and bristles â€“ no halos or contour lines
ï‚§ Preserves low contrast detail and avoids â€œstacking mushâ€ with deep stacks
o Fast and flexible retouching makes it easy to combine the best features of multiple algorithms as well as original frames
â€¢ Retouching supported by all versions, even at the lowest price
â€¢ Supports 8- and 16-bit input and output files
â€¢ Can generate stereo and 3-D rocking animations from a single stack, even for difficult subjects with structural overlaps and bristles.
â€¢ Fully utilizes modern multi-core processors and multi-processor computers
â€¢ Runs on all major computer systems: Microsoft WindowsÂ®, Macintosh OS XÂ®, LinuxÂ®.

Zerene Stacker license, as of 12/2013
â€¢ Professional Edition, $289 USD
â€¢ Prosumer Edition, $189 USD
â€¢ Personal Edition, $89 USD
â€¢ Student Edition, $39 USD

Ed

Edward Haller, Lab Manager
University of South Florida
Department of Integrative Biology
Electron Microscopy Core
SCA 110
4202 East Fowler Avenue
Tampa, FL 33620
(813)974-2676
ehaller-at-usf.edu
Office: ISA 1046
http://biology.usf.edu/ib/research/facilities/

________________________________________

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Email: scalliom-at-myumanitoba.ca
Name: Melanie Scallion

Organization: University of Manitoba

Title-Subject: [Filtered] SEM - Need help with which scope to use for insects

Message: I am an undergraduate student at the University of Manitoba working on a revision of a
genus of parasitic wasps. I have three new species that I would like to take images of in order to
see their sculpturing, and Iâ€™m trying to figure out which SEM I should use. The specimens are about
3mm in length, and because they are not mine, I need them to remain as is (they have to be returned
in the same condition in which I received them). The insects are point mounted, which means they are
glued to the end of a small, pointed piece of paper which then has a pin stuck through it. The glue
used likely dissolves in ethanol.

The two scopes we have here at the university are: Philips XL 30 and JEOL JSM-5900LV. Iâ€™m hoping
someone could provide me with a recommendation on which to use, and advise me on what I will need to
do if one of these scopes is suitable.

Any information would be of great help! Thank you,

Melanie Scallion

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Through the efforts of our Hitachi service engineer, we will have a needle valve installed tomorrow. Thanks to everyone for your helpful suggestions.

Dan Fairweather
Senior Materials Development Engineer
Advanced Materials, Dept. 32-22

-----Original Message-----
X-from: Philip Oshel [mailto:oshel1pe-at-cmich.edu]
Sent: Monday, June 30, 2014 11:29 AM
To: Fairweather, Dan

Years ago, we had done some work with Roy Larimer of MicroOptics. It looks like Visionary Digital is now selling his BK set-up.

http://www.visionarydigital.com/Pricing.html

The system was based on a clever use of image scanning. Also, he developed a special chamber for holding the bugs. The resulting bug images were amazing. I still have a copy of an app note we wrote and tried to locate the original website (2002!) but it now comes up in Japanese.

Check out the Visionary Digital site and see if this would be helpful.

Caveat: We have no commerical involvement/interest in this product.

Good Hunting!

Barbara Foster, President & Chief Consultant
Microscopy/Microscopy Education*
www.MicroscopyEducation.com

*A subsidiary of The Microscopy & Imaging Place, Inc.
7101 Royal Glen Trail, Suite A
McKinney, TX 75070
P: 972-924-5310
F: 214-592-0277

MME is currently scheduling courses for now and through Fall 2014. Call us today for a free training evaluation.

At 02:37 PM 6/29/2014, you wrote:

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From reasonable-meds10-at-charter.com Mon Jun 30 16:52:22 2014
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Barry;
Lucky you! At Intel, barriers to protect electron microscopes from sprinklers were not permitted, because if the microscope was on fire, the sprinklers had to put it out. The building was considered more important than the microscope!

John Mardinly, ASU

-----Original Message-----
X-from: barrylamb-at-meadtest.com [mailto:barrylamb-at-meadtest.com]
Sent: Friday, June 27, 2014 6:18 AM
To: John Mardinly

Hi Rosemary,

We had Perspex canopies made up for our SEMs, Auger system and SIMS machines to protect from direct sprinkler spray.

Regards, Barry

Barry Lamb
Director
Mead Testing Ltd
Registered address: 25 Mead Park,River Way, Harlow, Essex, CM20 2SE, UK Company no. 4572600 barrylamb-at-meadtest.com www.meadtest.com
tel: + 44 (0) 1279 635864
fax: +44 (0) 1279 635874

-----Original Message-----
X-from: protrain-at-emcourses.com [mailto:protrain-at-emcourses.com]
Sent: 27 June 2014 13:21
To: barrylamb-at-meadtest.com

Hi Rosemary

Out of interest I have dealt with water cascading over microscopes, yes
it
has happened twice in my 49 electron microscopy years. As a service
technician the main problem is not so much the water, but the debris
that
the water beings down onto the electrical circuits. Both of the
instruments
in question were running on vacuum only and managed to keep going
overnight
until security found the water running through the particular
laboratories.
One was a tap exploding, shooting a fountain of water into the EM room,
the
other the result of putting out a fire three levels above.

A third situation occurred when a water pipe came off one instrument in
a
basement EM unit, flooding the complete area to about 6 inches. Everyone
of
the instruments kept going, but as I arrived a technician with rubber
boots
and a long wooden pole was taking care of switching the most endangered
instruments off. However this was in the 1980s when most EM Units were
underground; the one I mentioned was also used as a Tornado shelter!

EM Units and water, great fun!

Steve

Steve Chapman FRMS
Protrain for Consultancy and Courses in Electron Microscopy
Mobile +44 (0) 7711 606967 Fax +44 (0)1280 814007
www.emcourses.com

-----Original Message-----
X-from: Rosemary.White-at-csiro.au [mailto:Rosemary.White-at-csiro.au]
Sent: 26 June 2014 11:09
To: protrain-at-emcourses.com

Dear Roger and Listers,

I have had the joy of a sprinkler system waterfall originally just outside
my TEM lab in the hallway a few years back. I say originally for the
water shortly flooded through the my space and several other labs and
offices. Hence the several inches of water was not over but under our
scope. Thankfully the scope was not damaged but the mess took several
days to clean up.

I have lived long enough that I DID experience a TEM full of water. It
was a Phillips 200 (1966 version) in the Biology Department at the
University of Pennsylvania. When I went on the scope and looked in the
binoculars it looked very strange. Water was in and on the outside of the
column. The water was not near the filament but I immediately turned it
off!!! One does not think to see if the column is flooded on a regular
basis. This was before Phillips became FEI but I do not remember the year,
maybe 1978? Two very talented service engineers spent about a week, if I
remember correctly, taking the column apart, drying each part out and
putting it back together. Several years later I found water on the outside
of the column in the crack where two lenses joined together but I caught
it at the beginning and shut down the water supply immediately. An O-ring
had broken where the hose was clamped into the lens, an easy fix. This old
scope outlived several of its service engineers and was turned off for the
last time in 2005.

Oh! I had also found that when my beam would pulse with a power
instability, it was not the fault of the filament or a bad vacuum tube or
a transistor but the SPARK PLUG. The very small gap would have a fleck of
carbon blocking it. After I ran a piece of paper through to open the gap
all would be fine for the next eight years or so.

Opinions and experiences related are those of Pat Connelly and do not
represent the NIH. This message is not confidential and
can be freely shared and reproduced.

Pat

Patricia Stranen Connelly
Research Assistant
NHLBI EM Core Facility
National Institutes of Health
Building 14E Room 111B
Bethesda, MD 20892-5570
301-496-3491

http://www.nhlbi.nih.gov/research/intramural/researchers/core/electron-micr
oscopy-core/index.html

==================
On 6/28/14 9:07 AM, "microscopylistserver-noreply-at-microscopy.com"
{microscopylistserver-noreply-at-microscopy.com} wrote:

} X-from: roger.ristau-at-uconn.edu ()
}
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} Email: roger.ristau-at-uconn.edu
} Name: Roger Ristau
}
} Organization: University of Connecticut
}
} Title-Subject: [Filtered] Re: Sprinklers in instrument rooms
}
} Message: Here is a situation that I bet not many have encountered: We had
} a TEM which developed a
} leak in lens cooling with the result that the INSIDE of the TEM filled
} with water. (I was not here
} but was told the camera/viewing chamber looked like "a fish bowl.")
}
} My predecessor, Larry McCurdy, courageously attacked the mess and
} restored the TEM to service! It
} soldiered on for many years.
}
} Point of story: Flooding the OUTSIDE of the TEM may actually be more
} harmful (electrical
} shorts,etc.) than flooding the INSIDE!
}
} Cheers
} Roger
}
} Institute of Materials Science
} University of Connecticut

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From drugs_reasonable14-at-rr.com Mon Jun 30 20:36:43 2014
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I wonder if anybody has explored the "Rocking" mode available in the ASID (scanning) panel on a JEM-2100 LaB6 or similar instrument. I can get nice channeling patterns using the backscatter detector; also some corresponding Kikuchi diffraction with the STEM detector. The software allows control of both working distance and camera length. But there is essentially no documentation on this beyond "Channeling pattern observation mode". It is not entirely clear to me what the actual ray diagram is here, or how it differs from normal STEM mode. Should the probe be rocking about a fixed point? The signal looks like a channeling pattern superimposed on an image. I'd especially like to do electron channeling contrast imaging.

Please let me know if you have any tips. Thanks.
------------------------------------------
Phil Ahrenkiel, Associate Professor
Nanoscience and Nanoengineering
South Dakota School of Mines and Technology
501 E. Saint Joseph St.
Rapid City, SD 57701 U.S.A.
Office: EP 221
Phone: 605-394-5238, Cell: 720-988-6627
Email: Phil.Ahrenkiel-at-sdsmt.edu

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5, 37 -- Subject: rocking on a JEM-2100
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Name: John Schreiber

Organization: Princeton Univ

Title-Subject: [Filtered] LAB6 filament

Message: Hello Rashmi,

You can check that the support of the LAB6 crystal is intact. Look at where the material that holds
the crystal in place is intact and does not have cracks. The cracks will cause beam instabilities.

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Title-Subject: [Filtered] Microscope Training

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Title-Subject: [Filtered] Detector info for DTSA II

Message: Has anyone had any luck obtaining detector info from Oxford to fill out the detector
preferences in DTSA II?

Thanks,

TJ

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Organization: Monash Centre for Electron Microscopy, Monash University

Title-Subject: [Filtered] Re: Two fixed-term scanning electron microscopist positions

Message: The Monash Centre for Electron Microscopy is seeking outstanding candidates for two
replacement Scanning Electron Microscopists. Both positions are located at the Clayton campus.

The Monash Centre for Electron Microscopy conducts research and provides advanced instrumentation,
training and expertise in electron microscopy. The Centre undertakes and facilitates research
projects across a range of disciplines in the physical sciences and engineering and serves
researchers from Monash University, other universities, government research agencies and industry.

The Centre has a world-class suite of instrumentation housed in a dedicated, high-stability building
designed to optimise instrument performance. You will provide expertise and training in scanning
electron microscopy to support the research activities of researchers using the Centre.

The successful applicants will have advanced understanding and skills in the acquisition and
interpretation of SEM data and its application to the solution of research problems in the physical
sciences. These positions are maternity leave replacements and will be for a minimum duration of 6
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Appointments will be made at the level appropriate to your experience and qualifications. Please
address the appropriate selection criteria according to your skills and qualifications.

These roles are full-time positions; however, flexible working arrangements may be negotiated.

For details see http://www.monash.edu.au/jobs/, ref. Electron Microscopist Job 525148

Enquiries: Professor Joanne Etheridge, Director, Monash Centre for Electron Microscopy, +61 3 9905
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Dear Dr White, my lab is also recently looking into building a new lab to house the Aberration Corrected TEM. We are also in the discussion whether we can do away the fire sprinkler. One proposal (which is not yet finalized) is to have a two-step solution. First is to have a pre-action system (plus sprinkler), second is to have a FM-200 waterless fire suppression systems. So if fire started,the FM-200 will be activated, after sometime if it didn't put out the fire, sprinkler will comes in (correct me if i am wrong in the procedure). However, in Smy country, we have various regulatories on this fire suppression system which we need to deal with before we consider viable. Another problem is the cost. So i am not sure if Australia allows.

Best Regards,
Yee Yan
FACTS Lab
Nanyang Technological University

On Sunday, 29 June 2014, 16:21, "Rosemary.White-at-csiro.au" {Rosemary.White-at-csiro.au} wrote:

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FYI a summary of responses. Thanks much to all, very useful.

1. Alternatives are available, but expensive. These include:
- firewalls around each room lacking sprinklers, and firedoor entryway -
rather expensive. The walls and door have to be able to resist fire for 1
hour.
- gas systems - also expensive, and in Australia these are apparently only
allowed in non-occupied rooms (I am told, not going to fight it, save
fight for other stuff...).

So

2. Accept the inevitability of sprinklers, and insist on the following:
- sprinkler head(s) as far from instrument as possible, esp. from main
power supplies
- sprinkler heads enclosed in wire cages to prevent accidental knocking
when moving large items in the room

3. Realise that floods from pipes/sinks/rain events are much more likely
than sprinkler activation, so:
- install drip pans above the sprinklers, either below the ceiling or
within it, to deflect potential floods from above away from the instrument
- raise important components at least several cm above floor level
- have a system that only fills the sprinkler pipes when the sprinkler(s)
are activated to avoid slow leaks (drips)

4. Also realise that sprinkler pipes may be sources of conductive
interference, so:
- ask that pipes are empty unless sprinkler(s) activated
- insert a rubber or other non-metallic connection between the main
sprinkler supply system and the sprinklers in instrument rooms

and also, check your insurance policy!

Like others, the only floods I've experienced were from other sources - in
a previous institution on one torrential weekend several staff came in to
check for leaks and ended up doing a bucket brigade from the EM unit in
the basement. All fine, except for the piles of paperwork around desks...

And Roger Ristau's email reminded me that about 15 years ago the in lens
cooling system in our JEOL 6400 developed a leak and flooded the lens,
detected by water dripping down into the chamber - messy! The old girl is
still chugging along, though not for much longer - the old electronic
boards are cracking and won't survive the move, so turning off for good in
October. :(

cheers,
Rosemary

Dr Rosemary White
CSIRO Plant Industry (for 1 more day)
GPO Box 1600
Canberra, ACT 2601
Australia

T 61 2 6246 5475
F 61 2 6246 5334
E rosemary.white-at-csiro.au

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Hi All,

I'd like to plastic embed samples of minced meat for LM and TEM analysis. My fear is that the mince will dissipate when put in glutaraldehyde for fixation. Do you have any experience or suggestion of how to keep the mince structure intact during the fixation, dehydration and infiltration steps?

Best regards,
Sofia

.............................
Dr Sofia Kihlman Øiseth
Research Scientist

CSIRO Food and Nutrition Flagship
E sofia.oiseth-at-csiro.au T +61 3 9731 3541
671 Sneydes road, Werribee, VIC 3030, Australia
www.csiro.au

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Organization: USDA ERRC

Title-Subject: [Filtered] denatured EtOH

Message: Is denatured ethanol sufficient for dehydrating samples for SEM & TEM? Or is absolute
required? There is quite a price difference, as well as ordering hurdles.

thanks

Joe

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Name: Phil Fraundorf

Organization: University of Missouri St. Louis

Title-Subject: [Filtered] TEM: javascript electron optics?

Message: Perhaps for the first time, HTML
with javascript FFT's is fast enough
on many platforms to do some
interesting stuff in real time. In
particular, we've been looking at
pedagogical uses for JS/HTML5 in
strong phase-object wave-propagation
& Fourier-window darkfield analysis.

This link* illustrates ways
these might be used e.g. to provide
microscopy students with on-line
exercises in focus-adjustment and
data-acquisition, as well as in
study of contrast-transfer and image
periodicities.

* https://sites.google.com/site/electrondetectives/tutorial-materials/f-javascript-fft-simulators

Unlike plugins for ImageJ,
routines in JS/HTML5 are not likely
to be much use for analyzing your
own data. Local programs are
generally needed for that. However
they do hold promise for giving folks
visceral experience with key microscopy
challenges. In this context, how might
freely-available web resources like
this be re-arranged so as to be
useful to folks learning to set up,
and analyze, images in your lab?

P. Fraundorf
UM-StL Physics & Astronomy/CNS

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From popular-medications8-at-detelic.com Thu Jul 3 07:09:48 2014
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Joe,
We run a large number of both TEM and SEM experiments and have always used
pint sized (open just before 100% step) Ethyl Alcohol 200 proof absolute
andydrous ACS/USP grade form Pharmco-AAPER #64-17-5. We use the opened
bottles to make ethanol parts and general lab use. It is crucial that the
ethanol contain no water. In the old days I used to molecular sieve the
ethanol, but it is no longer necessary.
Good luck;
Michael Delannoy

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Title-Subject: [Filtered] denatured EtOH

Message: Is denatured ethanol sufficient for dehydrating samples for SEM &
TEM? Or is absolute required? There is quite a price difference, as well
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thanks

Joe

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Hi Joe:
You should always dehydrate using pure ethanol, starting at 25 or 50%, then continue with these changes:
65%, 80%, 95%, 3x 100% then if you are using an epoxy resin transition into that.
Absolute ethanol is necessary for the 100% changes.
Karen Bentley
University of Rochester Medical Center

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Title-Subject: [Filtered] denatured EtOH

Message: Is denatured ethanol sufficient for dehydrating samples for SEM & TEM? Or is absolute
required? There is quite a price difference, as well as ordering hurdles.

thanks

Joe

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Title-Subject: [Filtered] opening RevolutionEDX files

Message: Greetings,

I have a user who collected TEM/EDS data on a 4pi RevolutionEDX system.
We need to (if possible) open and evaluate the spectrums. No access to
the RevEDX software itself....obviously, and the 4pi website wasn't much
help. Having no luck reading the files with software we have on hand.
I've tried DTSA-II with no luck??

Any suggestions or advice would be greatly appreciated.

Thanks!

Tom

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From reasonable-meds1-at-spbmts.ru Sun Jul 6 07:47:31 2014
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All,

Come to beautiful Tasmania for AMAS XIII, the 13th biennial Australian Microbeam Analysis Symposium, which will be held in Hobart on 11-13 February 2015, with Pre-Meeting Workshops 9-10 Feb, 2015.

The AMAS symposia provide a forum to discuss and share ideas on advances, trends and challenges in microanalysis and imaging with national and international leaders in the field, with an emphasis on practical solutions and applications.

Continuing the successful format of previous editions, AMAS XIII will be hosted at University venues and exclusively have oral presentations in a single session over three days, thus providing an intimate environment for in-depth discussions.

We invite you to present your research on technological developments and applications in a wide range of microscopy and microanalysis techniques.

A range of exciting invited speakers and workshops have already been confirmed.

For more information, see our website

http://microscopy.org.au/amas/amas13/

or talk to me or one of the other Aussies at M&M/IUMAS6 in Hartford.

Many thanks,

Karsten

AMAS XIII Co-Chair

Dr Karsten Goemann
Research Fellow, Electron Microscopy & X-Ray Microanalysis
Central Science Laboratory, University of Tasmania
Mail: Private Bag 74, Hobart TAS 7001, Australia
Location: Rooms 254-256, Chemistry Building, Dobson Road, Sandy Bay TAS 7005, Australia
T: +61 (0)3 6226 2146 | F: +61 (0)3 6226 2494 | M: +61 (0)407 101 990
www.utas.edu.au/research/central-science-laboratory
CRICOS 00586B

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From cheapest_drugstore7-at-in-addr.arpa Mon Jul 7 07:13:45 2014
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Getting the most from your AFM requires effective utilization of image
processing
software. This free, live-streaming seminar covers how to level images,

display images, and analyze images with AFM image processing software.
The demonstration will utilize Gwyddion's open source software and be
led
by Paul West, PhD.

To RSVP for this 30 minute AFM Image Processing Seminar, visit
AFMWorkshop's Ustream Channel:
http://www.ustream.tv/channel/Atomic-Force-Microscopes

Thank you,
Pamela Stone
AFMWorkshop, Inc.
1434 E. 33rd Street
Signal Hill, CA 90755
www.afmworkshop.com

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Hi, Everyone-

We have two Optronics cameras on light microscopes, a MacroFire and a
MagnaFire. One has a Sony chip and one a Kodak chip, and they have
different software, and we like both, especially Picture Frame
(MacroFire). But the MacroFire driver does not work past Windows XP
service pack 3, and the MagnaFire does not work past Win XP service pack
1. Their respective computers are dying, and although we keep putting them
on newer old computers, it is a losing battle. Has anyone come up with a
hack to keep the older Optronics cameras going? I can sort of make them
work with a generic twain driver, but we really like the capabiities of
Picture Frame for mixing channels, etc. I heard a rumor they would work on
Win 7 32 bit, but that didn't pan out. Any other solutions?

Aloha,
Tina

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

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4, 20 -- From tina-at-pbrc.hawaii.edu Tue Jul 8 18:34:55 2014
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From drugs-cheapest11-at-mts.ru Wed Jul 9 02:12:18 2014
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Hi Tina,

Have you tried Windows XP mode in Windows 7?
Odds are that the Windows 7 version you have access to allows for this
possibility.
Windows XP mode was designed to allow for the use of legacy software
in Windows 7. I am not sure how well it handles legacy hardware.

Some install info from Microsoft:
http://windows.microsoft.com/en-us/windows7/install-and-use-windows-xp-mode-in-windows-7

best of luck

Rafael Buono
Department of Botany
University of Wisconsin - Madison

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Group,

Interested in hearing from labs running EDS and WDS detectors on their SEM's.

Please comment on your experience with the hardware and software from vendor you are using.

Feel free to contact me directly with your thoughts.

Regards

Roy Beavers
rbeavers-at-smu.edu

==============================Original Headers==============================
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Hello listserv subscribers,

We're interested in purchasing an ion beam cross section polishing
system to complement our other SEM and TEM sample preparation
equipment. If you have experience with the systems from Gatan,
Hitachi, JEOL, or Leica, could you please contact me offline to
discuss the performance, reliability, ease of use, operating costs,
and any other useful information which will help guide our purchase.
Vendor responses are welcome as well.

Thank you for your help,
Chris
Senior Research Associate
Nanoscale Characterization and Fabrication Laboratory
Institute for Critical Technology and Applied Science
Virginia Tech
(540) 200-9511

==============================Original Headers==============================
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3, 26 -- Subject: Experiences with ion beam cross section polishing systems
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Dear Colleagues,

The diaphragm pump in our plasma cleaner (Gatan Model 950)is failing, we
are in need to find a replacement. This is a Pfeiffer MVP 020-3 DC
membrane pump, anyone has suggested vendors in US we can purchase from?

Thank you in advance.
Xinran Liu

________________________________________

Xinran Nick Liu, M.D. & Ph.D.
Director of CCMI Electron Microscopy Core Facility
Research Scientist in Cell Biology
Yale University School of Medicine
333 Cedar Street, SHM I-E16A
New Haven, CT 06520
Office: 203.785.4050
Lab: 203.785.5390
http://medicine.yale.edu/ccmi/em

==============================Original Headers==============================
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10, 35 -- From: "Liu, Xinran" {xinran.liu-at-yale.edu}
10, 35 -- To: "Microscopy-at-microscopy.com" {Microscopy-at-microscopy.com}
10, 35 -- Subject: Pump replacement for Gatan plasma cleaner
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Name: Rashmi

Organization: DBT

Title-Subject: [Filtered] Three way valve and filament

Message:
Dear All

Our 'FEI' HRTEM F-30 is down since over a month. There is a problem in 3
way valve in Zephyr Chiller. We would like hear from supplier who can
supply us on urgent basis.

FEI TEM T-20 is down as the filament is blown off. Can we get the
supplier in India who can supply the tungsten filament on urgent basis.

Regards
Rashmi

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University

Title-Subject: [Filtered] FEI service response time

Message: Hi - I was wondering what the response time has been for FEI
TEM users for service calls - average time for your service reps to show
up from the time you make a service call - thanks!!
Sue

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Dear Rashmi,

Sometimes a good hit with a hammer works on the three-way valve.
It can also be opened and cleaned.

Filaments can be ordered directly online at kymbalphysics.com.

Best,

Wim

} On Jul 10, 2014, at 19:49, zaluzec-at-microscopy.com wrote:
}
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} Name: Rashmi
}
} Organization: DBT
}
} Title-Subject: [Filtered] Three way valve and filament
}
} Message:
} Dear All
}
} Our 'FEI' HRTEM F-30 is down since over a month. There is a problem in 3
} way valve in Zephyr Chiller. We would like hear from supplier who can
} supply us on urgent basis.
}
} FEI TEM T-20 is down as the filament is blown off. Can we get the
} supplier in India who can supply the tungsten filament on urgent basis.
}
} Regards
} Rashmi
}
} Login Host: 14.139.126.167
} Listserver Email Form V - 20120416
} ---------------------------------------------------------------------------
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}
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Hi Sue,

Keep in mind that mere appearance of the representative does not yet
mean a solution to your problem. While Average Response Time (ART) is a
good indicator of responsiveness of the service organization, feedback
on Mean Time To Repair (MTTR) could be even more valuable in evaluating
its effectiveness.

When reviewing ART and MTTR from users and comparing there's feedback to
your situation, take a note of either response comes from a customer
under full "parts and labor" service contract or is it from Time and
Materials (T&M) service call. Regardless of the particular service
organization, wuite often there seem to be a significant difference in
availability and quality of service available to full-contract and T&M
customers.

Valery Ray
=================================
PBS&T, MEO Engineering Co., Inc.
290 Broadway, Suite 298
Methuen, MA 01844 USA
Phone: +1-978-296-5063 - leave a message with call-back number
US Mobile: +1-978-305-0479
Skype: pbstmeo
E-mail: vray-at-partbeamsystech.com
Web: www.partbeamsystech.com
Web: www.freudlabs.com

On 7/10/2014 1:35 PM, zaluzec-at-microscopy.com wrote:
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} Email: susan.vanhorn-at-stonybrook.edu
} Name: Susan C. Van Horn
}
} Organization: Central Microscopy Imaging Center (C-MIC) Stony Brook
} University
}
} Title-Subject: [Filtered] FEI service response time
}
} Message: Hi - I was wondering what the response time has been for FEI
} TEM users for service calls - average time for your service reps to show
} up from the time you make a service call - thanks!!
} Sue
}
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6, 37 -- Date: Thu, 10 Jul 2014 14:33:20 -0400
6, 37 -- From: Valery Ray {vray-at-partbeamsystech.com}
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6, 37 -- To: microscopy-at-microscopy.com, susan.vanhorn-at-stonybrook.edu
6, 37 -- Subject: Re: [Microscopy] viaWWW:FEI service response time
6, 37 -- References: {201407101735.s6AHZrsR031910-at-ns.microscopy.com}
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Contents Retrieved from Microscopy Listserver Archives
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Hi Rashmi,

In addition to the option of purchasing electron sources from either
Kimball Physics or Denka, you may consider refurbishing the old source
by replacing filament and the tip. You can contact York Probe Sources in
UK for tip rebuilding service.

Valery Ray
=================================
PBS&T, MEO Engineering Co., Inc.
290 Broadway, Suite 298
Methuen, MA 01844 USA
Phone: +1-978-296-5063 - leave a message with call-back number
US Mobile: +1-978-305-0479
Skype: pbstmeo
E-mail: vray-at-partbeamsystech.com
Web: www.partbeamsystech.com
Web: www.freudlabs.com

On 7/10/2014 2:05 PM, wim.hagen-at-me.com wrote:
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} Dear Rashmi,
}
} Sometimes a good hit with a hammer works on the three-way valve.
} It can also be opened and cleaned.
}
} Filaments can be ordered directly online at kymbalphysics.com.
}
} Best,
}
} Wim
}
} } On Jul 10, 2014, at 19:49, zaluzec-at-microscopy.com wrote:
} }
} }
} }
} }
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} } Email: rashmi_mehata-at-yahoo.com
} } Name: Rashmi
} }
} } Organization: DBT
} }
} } Title-Subject: [Filtered] Three way valve and filament
} }
} } Message:
} } Dear All
} }
} } Our 'FEI' HRTEM F-30 is down since over a month. There is a problem in 3
} } way valve in Zephyr Chiller. We would like hear from supplier who can
} } supply us on urgent basis.
} }
} } FEI TEM T-20 is down as the filament is blown off. Can we get the
} } supplier in India who can supply the tungsten filament on urgent basis.
} }
} } Regards
} } Rashmi
} }
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4, 44 -- Date: Thu, 10 Jul 2014 14:44:16 -0400
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4, 44 -- Subject: Re: [Microscopy] Re: viaWWW:Three way valve and filament for FEI
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Contents Retrieved from Microscopy Listserver Archives
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Dear All,

The Microscopy and Microanalysis Meeting (Aug. 3-7 2014 in Hartford, CT) is
coming closer and I would like to make you aware that there are still openings
in our Sunday Short Course on 3D electron microscopy (description below).It
is not too late to register for it.

This course is ideal for people who need a comprehensive introduction or a
refresher in 3D reconstruction methods and is suitable for biologists and
material scientists. If you have already registered for the meeting you still
can add the course registration as "extra item only".

Description:

X12 3D Electron Microscopy of Macromolecular Assemblies
Teresa Ruiz, Michael Radermacher, Edward Morris
8:30 AM to 5:00 PM on Sunday, August 3.

Sample preparation: deep stain, vitreous ice
Imaging conditions, low-dose imaging, tilt-pair data collection
Alignment techniques and multivariate statistical analysis
3D reconstruction methods
X-ray structure docking
Techniques described have applications in both biological and
material science

This short course will provide a comprehensive description of the methods used
for 3D structure determination from electron micrographs of macromolecular
complexes or weakly scattering specimens available in multiple
copies.Specimen-
preparation techniques for single particles (deep stain, vitreous ice) will be
presented,followed by selection of optimal imaging conditions, including
low-dose imaging. Next,a detailed explanation of image-processing techniques,
with special emphasis on the random-conical reconstruction technique, will be
presented. Finally, structure interpretation and docking of X-ray structures
to 3D EM densities will be demonstrated.The techniques described could be
applied to both biological and materials science specimens.

--
Michael Radermacher, Ph.D., Prof., FMSA
University of Vermont
Dept. Molecular Physiology and Biophysics
HSRF 120 / 149 Beaumont Ave
Burlington, VT 05405
USA

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Title-Subject: [Filtered] Radioisotopes in the TEM (Ga-67)

Message: We've received a request for imaging Ga67-containing particles
in the TEM. Our facility has very little experience with radioisotopes.

Ga67 is a gamma emitter, with a 1/2 life of 3.6 days. The Ga is
supposedly bound within the matrix of the particles, but I do not have
the details of the kind of material in which the Ga is embedded. The
dose on a single grid works out to be around 2 nCi. We'd most likely
have to image 1-2 grids.

What pitfalls and precautions should be considered when dealing with
this kind of sample, if we choose to handle it? My main concerns are Ga
migration, cross-contamination, and personnel safety.

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Title-Subject: [Filtered] annealing nanoparticles on c-film grids

Message: Has anyone annealed nanoparticles on TEM c-film grids? If so,
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Dear Listers

I was wondering if any of you could help me out. We have recently
installed a new SDD on our Jeol 1200 mk2 TEM it has a ASID with a
SAP10 polepice and it is mounted on the high angle 70 degree port. We
have a standard tungsten source in this one. The problem we have is we
are getting a lot less stored counts with the new detector compared to
the old SiLi detector, and we are picking up more noise and stray. We
have rulled out hardward, software, and site but are now looking at
geometry.

I was wondering if anyone out there who has the same TEM
(with/without ASID) with EDS in the same port could let me know make,
model, toa, crystal tilt and flange angle or any similar experiences
it would be very useful. if you have a rough idea of counts at a given
setup that would also be really helpful.

Thanks in advance
John

==============================Original Headers==============================
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From bimyekno-at-net.upcbroadband.cz Fri Jul 11 08:07:46 2014
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Dear Marissa,

unfortunately I can't help with this your request anyway.

But in case you are also on the platform {ResearchGate}
http://www.researchgate.net/

you should enter the Link/Address (= search phrase):
https://www.researchgate.net/search.Search.html?query=Annealing%20nanoparticles&type=question
an you'll get a lot of threads (Questions and answers) dealing with {annealing...} , some of them also of {nanoparticles}

Best wishes, good luck and have a nice and recreative weekend,

Wolfgang

Wolfgang MUSS PhD
Member of MSA
EM-Lab, Pathology
SALK-LKH & PMU SALZBURG
AUSTRIA

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} An: Muß Wolfgang
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} Organization: NCEM, LBNL
} Title-Subject: annealing nanoparticles on c-film
} grids
} Message:
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} Has anyone annealed nanoparticles on TEM c-film grids? If so, can you
} tell me the temp and time at which the carbon started to break
} down?
} And the specific grid you used for the experiment?
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Our first annual student poster and speaker competition will be held at the Frederick Seitz Materials Research Laboratory this coming November at our third annual Biological Structures workshop.

This is a conference covering biological imaging, or a combination of biological and materials imaging.

November 12-13th
Winners will receive $100 check and their image and names in local papers and on our web site.

In order to give Researchers time to allow their students to prepare, we already have all the rules, regulations and registration up and ready for participants at our online site.

http://mrl.illinois.edu/events/conferences-workshops/third-annual-biological-imaging-workshop

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

Vendors!

Your registration is also ready for the workshop, and includes the ability to help contribute to the contest prize ( email Lou Ann at lamiller-at-illinois.edu for details and if such things are yet filled)
or help be a pizza sponsor for the second day ( able to do at registration).

Those vendors offering to help with the prize will be limited to $100 ( state & University regulations), will be able to name the prize after your company, and will be included in judging and images that go to the local papers and online.

We are still formulating our tutorial speaker program, that will be posted as soon as we have it. Vendors, there is an opportunity to give talks, they must be tutorial in nature. If time permits 45 min, but be prepared for a 30 min slot. A third day may be set up where all can come for free if the numbers of presentations permit.

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

Our facilities has a PC, slots for USB, laptop connection( Mac & PC), laser pointers and lapel microphone.
We have poster size regulations in the poster competition rules, speaker rules are also online.

{ { { { { { { { { { { { { { { { {} } } } } } } } } } } } } } } }
Lou Ann Miller
Biological Electron Microscopy
Frederick Seitz Materials Research Laboratory
Lab Room 125 Office 374 MRL
104 South Goodwin Ave
Urbana, IL 61801
Campus mail code: MC-230

217-244-1567
lamiller-at-illinois.edu
http://mrl.illinois.edu

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From drugs-cheapest1-at-marveldirectory.com Fri Jul 11 13:27:13 2014
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Dear Marissa,

Carbon films are stable in the TEM vacuum up to 1000°C and even far
beyond. But the choice of the film supporting grid is critical.

I used with success gold 200 mesh grids with thin carbon films during my
PhD work on the melting temperature of gold nanocrystals (Buffat,
Phys.Rev. A13 (1976) 2287).

A priori molybdenum grids should had allowed high temperatures with good
mechanical stability. But it rapidly appeared that the C-film reacts
with Mo to form Mo carbide and cracks (see for instance Leroy et al,
J.Appl.Phys 99 (2006) 063704 about Mo-C reactivity).
Copper grids were excluded because of the risk of alloying with gold.
Eventually gold grids were the best compromise though the largest Au
crystals did melt only a few degrees before the supporting grid itself.

The vacuum in the electron diffraction camera (a TEM without magnifiying
section) was 2 10-6 mbar. A slow heating cycle took about 15 minutes and
I didn't observed any significant damage to the film when the sample was
transferred to a TEM for crystal size observation.

More experiments were performed with heating in a TEM (Philips EM430)
though with less temperature measurement accuracy, but still no
significant film damage due to heating.

However it appeared that the carbon film may destroys under strong
electron beam during HRTEM observation. At that time we still used
photographic plan films and we noticed a clear correlation between the
film sputtering speed and the time between loading films in the camera
and HRTEM observation. This shows that despite the presence of a
differential pressure aperture between the plate camera and the TEM
column itself, water (or methane?) molecules were able to fly straight
toward the sample and enhance sputtering (the mean free path of
molecules is about meter(s) in this vacuum range). Film stability under
strong electron beams depends more on the vacuum quality and residual
atmosphere composition than temperature.

Regards

Philippe

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10, 19 -- From philippe.buffat-at-epfl.ch Sat Jul 12 08:39:14 2014
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From medications-reasonable7-at-thrane.com Sat Jul 12 18:18:13 2014
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Dear all,

I am studying cellulose aerogel made from Nanocellulose Fibres (10-30nm diameter and several micron in length).

MATERIAL:
The material show highly porosity (up to 90% air) and a very intricate structure with meso and nano pores.

SAMPLE:
Samples are produced via supercritical CO2 drying after filtration and solvent exchange. The resulting sample is a ""paper-like"" fluffy material, 3 cm in diameter and a thickness of 300 micron.

SEM:
To study the microstructure I tried to embed a small piece of aerogel in araldite. The cross-section surface of the sample was exposed with a diamond microtome and sputtered with 20nm of gold.

In the obtained images is difficult to distinguish between the araldite matrix and the sample (a small change in contrast at low magnification). Worst is that on the surface I don't see any micro-structure. Only a light grey smooth surface.

QUESTION:
- Is it possible in your opinion to obtain a nice cross section image of my aerogel?
- I am thinking to change the embedding material with a more stiff epoxy. It could help?
- Do you have any others ideas?

Many thanks in advance,
Ferruccio

PS: This is my first message. Hope I did everything correct.

==============================Original Headers==============================
11, 41 -- From Ferruccio.Bolla-at-empa.ch Mon Jul 14 01:18:38 2014
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Hi,
Can you explain why you want to embed your sample? Particularly if
you can coat with something finer than gold (platinum, carbon) and if
you have access to some low voltage/charge minimizing features of the
recent generation of SEMs, it should work. Just a thought.

Tobias Baskin

On 7/14/14, 2:19 AM, Ferruccio.Bolla-at-empa.ch wrote:
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}
} Dear all,
}
} I am studying cellulose aerogel made from Nanocellulose Fibres (10-30nm diameter and several micron in length).
}
} MATERIAL:
} The material show highly porosity (up to 90% air) and a very intricate structure with meso and nano pores.
}
} SAMPLE:
} Samples are produced via supercritical CO2 drying after filtration and solvent exchange. The resulting sample is a ""paper-like"" fluffy material, 3 cm in diameter and a thickness of 300 micron.
}
} SEM:
} To study the microstructure I tried to embed a small piece of aerogel in araldite. The cross-section surface of the sample was exposed with a diamond microtome and sputtered with 20nm of gold.
}
} In the obtained images is difficult to distinguish between the araldite matrix and the sample (a small change in contrast at low magnification). Worst is that on the surface I don't see any micro-structure. Only a light grey smooth surface.
}
} QUESTION:
} - Is it possible in your opinion to obtain a nice cross section image of my aerogel?
} - I am thinking to change the embedding material with a more stiff epoxy. It could help?
} - Do you have any others ideas?
}
} Many thanks in advance,
} Ferruccio
}
}
} PS: This is my first message. Hope I did everything correct.
}
}
} ==============================Original Headers==============================
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--
__ ___ ^ ___ ___ Tobias I. Baskin
/ \ / / \ / \ Professor
/ / / / \ \ \ Biology Department
/ __/ /__ /___ \ \ \__ 611 N. Pleasant St.
/ / / \ \ \ University of Massachusetts
/ / / \ \ \ Amherst, MA, 01003 USA
/ /___ / \ \___/ \_____ 413 - 545 - 1533
www.bio.umass.edu/biology/baskin

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Ferrucio,

Why embed the aerogel? I've done aerogel in the SEM, but never with
embedding.
Coat with something other than plain gold - Au/Pd, Pt, or something -
and use low kV. "Low kV" depends on what SEM you have, but no more than
5 kV. If you can go down to 1 or 1.5 kV, that would be better. Possibly
lower. If you can find the kV at which the charges on the sample
balance, you may not need coating at all.
Also, you can submerge the aerogel in liquid nitrogen and carefully snap
it to expose the interior. I'm assuming the fibers have glass transition
above the LN2 temperature, but I'm sure it does.

If you have to embed the nanocellulose fibers in Araldite, you can stain
the Araldite with RuO4: place the sectioned blocks in a sealed dish in a
fume hood with a crystal or two of RuO4, and let the vapors stain the
resin. The nanocellulose should remain unstained.
Or: sputter coat the aerogel with gold or any heavy metal, but use
shorter coating runs, and turn over the block of aerogel between coats,
so the metal gets equal access to the aerogel from all sides. Then
embed. The fibers will be the bits that are surronded by the heavy
metal, which will show up in the SEM. Mind, this last is only good for
sections of the fibers, the surface will be obscured.

Phil

} Dear all,
}
} I am studying cellulose aerogel made from Nanocellulose Fibres (10-30nm diameter and several micron in length).
}
} MATERIAL:
} The material show highly porosity (up to 90% air) and a very intricate structure with meso and nano pores.
}
} SAMPLE:
} Samples are produced via supercritical CO2 drying after filtration and solvent exchange. The resulting sample is a ""paper-like"" fluffy material, 3 cm in diameter and a thickness of 300 micron.
}
} SEM:
} To study the microstructure I tried to embed a small piece of aerogel in araldite. The cross-section surface of the sample was exposed with a diamond microtome and sputtered with 20nm of gold.
}
} In the obtained images is difficult to distinguish between the araldite matrix and the sample (a small change in contrast at low magnification). Worst is that on the surface I don't see any micro-structure. Only a light grey smooth surface.
}
} QUESTION:
} - Is it possible in your opinion to obtain a nice cross section image of my aerogel?
} - I am thinking to change the embedding material with a more stiff epoxy. It could help?
} - Do you have any others ideas?
}
} Many thanks in advance,
} Ferruccio
} PS: This is my first message. Hope I did everything correct.

--
Philip Oshel
Microscopy Facility Supervisor
Biology Department
024C Brooks Hall
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576

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Hi Ferruccio
If you don't manage to manually open the aerogel successfully, you can try
to embed it paraffine and get some thick sections, which then you treat with
a warm (} 70C) solven compatible with your staff to get rid of the wax and
then coat and view in SEM?
best
yorgos

----- Original Message -----
X-from: "Bolla, Ferruccio" {Ferruccio.Bolla-at-empa.ch}
To: "yorgos nikas" {eikonika-at-otenet.gr}
Sent: Monday, July 14, 2014 12:46 PM

Hi Ferruccio,
it might be the same problem like cutting polystyrene (styropor).
I succeeded only with a piece of polystyrene submerged in liquid nitrogen and then using a NEW razorblade cooled at LN2 temp to
cut the polystyrene. The cells and walls looked relatively unharmed in the SEM afterwards.
See some images of it here:
http://www.electronmicroscopy.info/shop_materials.htm
image number 27 to 31...

Best wishes,
Stefan

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Arndtstrasse 22
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Websites:
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Anfahrt: http://Mail.map24.com/Stefan.Diller
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Am 14.07.14 08:24, schrieb Ferruccio.Bolla-at-empa.ch:
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}
} Dear all,
}
} I am studying cellulose aerogel made from Nanocellulose Fibres (10-30nm diameter and several micron in length).
}
} MATERIAL:
} The material show highly porosity (up to 90% air) and a very intricate structure with meso and nano pores.
}
} SAMPLE:
} Samples are produced via supercritical CO2 drying after filtration and solvent exchange. The resulting sample is a ""paper-like"" fluffy material, 3 cm in diameter and a thickness of 300 micron.
}
} SEM:
} To study the microstructure I tried to embed a small piece of aerogel in araldite. The cross-section surface of the sample was exposed with a diamond microtome and sputtered with 20nm of gold.
}
} In the obtained images is difficult to distinguish between the araldite matrix and the sample (a small change in contrast at low magnification). Worst is that on the surface I don't see any micro-structure. Only a light grey smooth surface.
}
} QUESTION:
} - Is it possible in your opinion to obtain a nice cross section image of my aerogel?
} - I am thinking to change the embedding material with a more stiff epoxy. It could help?
} - Do you have any others ideas?
}
} Many thanks in advance,
} Ferruccio
}
}
} PS: This is my first message. Hope I did everything correct.
}
}
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Dear All, recently, i am trying to help a user to section her sample embedded in Epo-Fix epoxy. As i am not very experience with ultramicrotome, I decide to section the Epo-Fix epoxy without any sample in it with a glass knife as a start. Unfortunately , i am unable to section it to a thickness of about 100nm. What i observed is that the glass knife is only able to section at alternating interval. I learn that this can be due to either the epoxy is quite soft of the glass knife is not sharp enough (or the block face might still be too big?). The student told me that that glass transition of the epoxy is above room temperature so there shouldn't be a need to adopt cryo-ultramicrotomy approach. I also attempt to trim the block face to a small size e.g to about 1mm (or slightly less) but still the same alternate sectioning observation is made. I have tried sectioning with biological sample embedded in aradite before and there is no problem achieving
thickness of 100nm or less.

My glass knife cutting angle is at 6 degree.

May i ask if i have missed out anything? Is Epo-fix suitable to be sectioned with a glass knife to achieve a thickness of 100nm? On my second question, does the Tg of the sample determines if one should use a room-temperature ultramicrotome or a cryo-ultramicrotome?

Cheers,
Yee Yan, Tay
FACTS Lab
Nanyang Technological University

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Dear Yee Yan Tay,

IMHO we need also to know which ultramicrotome (thermal or mechanical feed) you are using for this approach.
Epo-Fix is known to me as a specimen mountant(mounting medium) in Materials Sciences
(cf. e.g. http://www.struers.com/resources/elements/12/255648/Cold%20Mounting%20table_Epoxies.pdf )
and unfortunately I haven't experience with sectioning Epo-Fix-Epoxy embedded specimens. *)
It might be this resin too soft or too hard. It might be also the type of glass strips you use and how the quality of your knives really is.
Trials to overcome the problem with your glass knives (including the existing angle to be 35-45-55 degr. comparable with the diamond knives 35/45/55 degr.**)) could be:
i) sectioning at a (narrower) cutting angle of say 4.5 - 5.5 degree (instead of using 6 degr.),
ii) lowering level of water in the knife boat
iii) playing around with section speed (ensuring that the section phase is set sufficiently long enough, i. e. start cutting at least 1 mm above block edge, end of cutting phase as short as possible below /after having sectioned through the block at its lower edge).
iv) trying to harden the block further (oven over night, 65-80 degr.C.)
v) last but not least - only if available - use of a diamond knife.

With the usual resins for TEM (e.g. Epon 812, Embed812, Glycidether 100 substitute (for Epon 812), LX-112 etc in former times (also using glass knives) - when mixed thoroughly according to the recommandations of LUFT (1961) and others I had no problems cutting also bigger specs at least up to 4x4 mm (for my thesis I had a really soft resin mixture for cutting whole rat hypothalami - 1day PN to 320 days of age - including the ventricle).

Would like to ask you honestly to keep us informed about your problem and eventually the solution you found (or even personal Re's you receive on this unfortunate but interesting matter),
best of luck,
regards
Wolfgang

Wolfgang MUSS PhD
EM-Lab, Pathology
SALK-LKH & PMU Salzburg
SALZBURG, Austria
[if you need the pdf and can't download, let me know and I will send it to you -at- (rongchigram79-at-yahoo.com.sg)]

*) From: EPOFIX AND VACUUM: AN EASY METHOD TO MAKE CASTS OF HARD SUBSTRATES
Jan Kresten Nielsen and Jesper Maiboe
[Nielsen, Jan Kresten and Maiboe, Jesper 2000. Epofix and Vacuum: An Easy Method to Make Casts of Hard Substrates.
Palaeontologia Electronica, vol. 3, issue 1, art. 2: 10pp., 1.3MB.
http://palaeo-electronica.org/2000_1/epofix/issue1_00.htm
Copyright: Palaeontological Association, 15 April 2000
Submission: 20 August 1999, Acceptance: 24 February 2000]
see: Palaeontologia Electronica-http://www-odp.tamu.edu/paleo
"The sectioning properties of Epofix mixture are good. A comparison with other embedding media is beyond the scope of this paper. The reader is referred to Bromage (1985), who provided a systematic evaluating procedure for casts and moulds."
(Bromage, T. G. 1985. Systematic inquiry in tests of negative/positive replica combinations for SEM. Journal of Microscopy, 137:209-216)

**) cf. http://cime.epfl.ch/files/content/sites/cime2/files/shared/Files/Teaching/MSE_603_2013_autumn/Chapter%2012%20Sample%20prep%202013.pdf
(pdf of ppt: TEM Samples preparation_D.Laub_2013 ) scroll down to foil 75:
Resin: Knive angle Compression
Epofix 45° 11%
35° 6%
the further (following foils) 76ff perhaps also are interesting....Foil 80: Section thickness 40 - 50nm; Sectioning speed 0.2mm/sec

Searching GOOGLE for { EPOfix AND cutting quality sectioning for TEM or Transmission electron microscopy } reveals 241 results.

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

Von: rongchigram79-at-yahoo.com.sg [mailto:rongchigram79-at-yahoo.com.sg]
Gesendet: Montag, 14. Juli 2014 17:54
An: Muß Wolfgang
Betreff: [Microscopy] Experience with Epo-Fix Epoxy as Embedding Medium for Ultramicrotome
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Dear All, recently, i am trying to help a user to section her sample embedded in Epo-Fix epoxy.
As i am not very experienced with ultramicrotome, I decided to section the Epo-Fix epoxy without any sample in it with a glass knife as a start.

Unfortunately, i am unable to section it to a thickness of about 100nm.
What i observed is that the glass knife is only able to section at alternating interval.
I learn that this can be due to either the epoxy is quite soft of [or] the glass knife is not sharp enough (or the block face might still be too big?).

The student told me that that glass transition of the epoxy is above room temperature so there shouldn't be a need to adopt cryo-ultramicrotomy approach. I also attempt to trim the block face to a small size e. g to about 1mm (or slightly less) but still the same alternate sectioning observation is made. I have tried sectioning with biological sample embedded in araldite before and there is no problem achieving thickness of 100nm or less.

My glass knife cutting angle is at 6 degree.

May i ask if i have missed out anything?
Is Epo-fix suitable to be sectioned with a glass knife to achieve a thickness of 100nm?
On my second question, does the Tg of the sample determine if one should use a room-temperature ultramicrotome or a cryo-ultramicrotome?

Cheers,
Yee Yan, Tay
FACTS Lab
Nanyang Technological University

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Hi Ferruccio,

Welcome to the list!

I have done quite a bit of imaging of SiO2 aerogel, (not nanocellulose) but some of our imaging approaches may work for you.

To image it in an SEM, I use low voltage, uncoated. Â Because of the low density of aerogel, the charge balance point is surprisingly high. Â Our aerogels are especially low density aerogels (about 20 mg/cm3, which for SiO2 is about 99.3% vacuum) and the charge point on most of our aerogels comes in at around 2.1 to 2.2 keV. Â You can get some very nice images this way. It helps to use a field emitter because of the very fine structure of the fibers, and the fact that you want to keep the current low as well as the voltage.

For sectioning aerogel, we use motorized glass needle cutting: Â Westphal, A. J., Snead, C., Butterworth, A. L., Graham, G. A., Bradley, J. P., Bajt, S., et al. (2004). Aerogel keystones: Extraction of complete hypervelocity impact events from aerogel collectors. Meteoritics and Planetary Science, 39(8), 1375â€“1386.

This may be a lot of work to set up, but it works very, very well. Â One thing you can do is cut a 100 micron thick section, flip it on its side, and cut another section a few tens of microns thick. Â Then you can affix that to a substrate and image it uncoated with high voltage in an SEM. Â It works because the density is so low the electron beam just goes clean through. Â (Poor man's TEM.)

Finally, I have gotten images in a TEM, but since I'm not usually studying the aerogel itself (but rather minerals trapped in it) I've never developed a good protocol here. Â However, it is my experience that epoxy embedding always alters the structure of the aerogel, so you probably will be better off going with some of the other suggestions if the above doesn't work for you.

I concur with Stephan that it is possible to get a good cut with razorblades (though don't drink coffee first -- it will make your hand shake!) Â And Phil's approach seems sharp too.

Hope this helps,

Zack

}
} } Ferruccio.Bolla-at-empa.ch:
} } ----------------------------------------------------------------------------
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} } ----------------------------------------------------------------------------
} }
} } Dear all,
} }
} } I am studying cellulose aerogel made from Nanocellulose Fibres (10-30nm diameter and several micron in length).
} }
} } MATERIAL:
} } The material show highly porosity (up to 90% air) and a very intricate structure with meso and nano pores.
} }
} } SAMPLE:
} } Samples are produced via supercritical CO2 drying after filtration and solvent exchange. The resulting sample is a ""paper-like"" fluffy material, 3 cm in diameter and a thickness of 300 micron.
} }
} } SEM:
} } To study the microstructure I tried to embed a small piece of aerogel in araldite. The cross-section surface of the sample was exposed with a diamond microtome and sputtered with 20nm of gold.
} }
} } In the obtained images is difficult to distinguish between the araldite matrix and the sample (a small change in contrast at low magnification). Worst is that on the surface I don't see any micro-structure. Only a light grey smooth surface.
} }
} } QUESTION:
} } - Is it possible in your opinion to obtain a nice cross section image of my aerogel?
} } - I am thinking to change the embedding material with a more stiff epoxy. It could help?
} } - Do you have any others ideas?
} }
} } Many thanks in advance,
} } Ferruccio
} }
} }
} } PS: This is my first message. Hope I did everything correct.
} }

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Dear List
In our Jeol JSEM 5600LV the filament current does not increase and after the
scroll bar reaches the middle a message appears "filament burn out". Yet the
filament is not burned and we can see an instant glow when the high tension
is turned on.
The last days the "filament burn out" message was coming on with increasing
frequency and we had to turn on again and again the high tension to continue
working.
The Wehlnet cap and the connections of the high tension cable on the top of
the gun look fine, so far I can see
Any suggestion will be greatly appreciated
Best regards

yorgos

Dr Yorgos Nikas
Athens Innovative Microscopy
Skra 36 Voula 16673 GREECE

Tel/fax +30 210 8957677
mobile +30 6945 107477
www.eikonika.net www.aim.cat
************************************

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Hi

We recently observed some polymer aerogels in un test run for the
purchase of a new SEM, and with recent SEMs it is possible to do nice
things at very low voltage (300 V) in deceleration mode (the sample is
positively biased to slow down the incident electrons, and the SE and BE
are re-accelerated up to the detector), without any coating. It works
well, with imaging up to more than 100 kx, but need some skill to find
the right conditions.

For the cross-section, why not try the ion beam cross-polisher. You
could obtain some test run asking one of the manufacturer (Gatan, Jeol,
Hitachi, Leica Technor-Linda, etc). No embeding, possibily no metalisation.

Hope it helps !

Jacques

Le 14/07/2014 08:31, Ferruccio.Bolla-at-empa.ch a écrit :
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
} To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
} On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} ----------------------------------------------------------------------------
}
} Dear all,
}
} I am studying cellulose aerogel made from Nanocellulose Fibres (10-30nm diameter and several micron in length).
}
} MATERIAL:
} The material show highly porosity (up to 90% air) and a very intricate structure with meso and nano pores.
}
} SAMPLE:
} Samples are produced via supercritical CO2 drying after filtration and solvent exchange. The resulting sample is a ""paper-like"" fluffy material, 3 cm in diameter and a thickness of 300 micron.
}
} SEM:
} To study the microstructure I tried to embed a small piece of aerogel in araldite. The cross-section surface of the sample was exposed with a diamond microtome and sputtered with 20nm of gold.
}
} In the obtained images is difficult to distinguish between the araldite matrix and the sample (a small change in contrast at low magnification). Worst is that on the surface I don't see any micro-structure. Only a light grey smooth surface.
}
} QUESTION:
} - Is it possible in your opinion to obtain a nice cross section image of my aerogel?
} - I am thinking to change the embedding material with a more stiff epoxy. It could help?
} - Do you have any others ideas?
}
} Many thanks in advance,
} Ferruccio
}
}
} PS: This is my first message. Hope I did everything correct.
}
}
} ==============================Original Headers==============================
} 11, 41 -- From Ferruccio.Bolla-at-empa.ch Mon Jul 14 01:18:38 2014
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--

J. Faerber
IPCMS-DSI
Institut de Physique et Chimie des Matériaux de Strasbourg
Département Surfaces et Interfaces
23, rue de Loess ; BP43
67034 Strasbourg CEDEX 2
France

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

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Contents Retrieved from Microscopy Listserver Archives
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Dr Muss, sorry to add in the information. i am using Leica UCT. I use a mechanical feed of 100nm. At 300nm, it seems not a problem.

Cheers,
Yee Yan

On Tuesday, 15 July 2014, 1:16, "W.Muss-at-salk.at" {W.Muss-at-salk.at} wrote:

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Dear Yee Yan Tay,

IMHO we need also to know which ultramicrotome (thermal or mechanical feed) you are using for this approach.
Epo-Fix is known to me as a specimen mountant(mounting medium) in Materials Sciences
(cf. e.g. http://www.struers.com/resources/elements/12/255648/Cold%20Mounting%20table_Epoxies.pdf )
and unfortunately I haven't experience with sectioning Epo-Fix-Epoxy embedded specimens. *)
It might be this resin too soft or too hard. It might be also the type of glass strips you use and how the quality of your knives really is.
Trials to overcome the problem with your glass knives (including the existing angle to be 35-45-55 degr. comparable with the diamond knives 35/45/55 degr.**)) could be:
i) sectioning at a (narrower) cutting angle of say 4.5 - 5.5 degree (instead of using 6 degr.),
ii) lowering level of water in the knife boat
iii) playing around with section speed (ensuring that the section phase is set sufficiently long enough, i. e. start cutting at least 1 mm above block edge, end of cutting phase as short as possible below /after having sectioned through the block at its lower edge).
iv) trying to harden the block further (oven over night, 65-80 degr.C.)
v) last but not least - only if available - use of a diamond knife.

With the usual resins for TEM (e.g. Epon 812, Embed812, Glycidether 100 substitute (for Epon 812), LX-112 etc in former times (also using glass knives) - when mixed thoroughly according to the recommandations of LUFT (1961) and others I had no problems cutting also bigger specs at least up to 4x4 mm (for my thesis I had a really soft resin mixture for cutting whole rat hypothalami - 1day PN to 320 days of age - including the ventricle).

Would like to ask you honestly to keep us informed about your problem and eventually the solution you found (or even personal Re's you receive on this unfortunate but interesting matter),
best of luck,
regards
Wolfgang

Wolfgang MUSS PhD
EM-Lab, Pathology
SALK-LKH & PMU Salzburg
SALZBURG, Austria
[if you need the pdf and can't download, let me know and I will send it to you -at- (rongchigram79-at-yahoo.com.sg)]

*) From: EPOFIX AND VACUUM: AN EASY METHOD TO MAKE CASTS OF HARD SUBSTRATES
Jan Kresten Nielsen and Jesper Maiboe
[Nielsen, Jan Kresten and Maiboe, Jesper 2000. Epofix and Vacuum: An Easy Method to Make Casts of Hard Substrates.
Palaeontologia Electronica, vol. 3, issue 1, art. 2: 10pp., 1.3MB.
http://palaeo-electronica.org/2000_1/epofix/issue1_00.htm
Copyright: Palaeontological Association, 15 April 2000
Submission: 20 August 1999, Acceptance: 24 February 2000]
see: Palaeontologia Electronica-http://www-odp.tamu.edu/paleo
"The sectioning properties of Epofix mixture are good. A comparison with other embedding media is beyond the scope of this paper. The reader is referred to Bromage (1985), who provided a systematic evaluating procedure for casts and moulds."
(Bromage, T. G. 1985. Systematic inquiry in tests of negative/positive replica combinations for SEM. Journal of Microscopy, 137:209-216)

**) cf. http://cime.epfl.ch/files/content/sites/cime2/files/shared/Files/Teaching/MSE_603_2013_autumn/Chapter%2012%20Sample%20prep%202013.pdf
(pdf of ppt: TEM Samples preparation_D.Laub_2013 ) scroll down to foil 75:
Resin:     Knive angle Compression
Epofix     45°    11%
        35°    6%
the further (following foils) 76ff perhaps also are interesting....Foil 80: Section thickness 40 - 50nm; Sectioning speed 0.2mm/sec

Searching GOOGLE for { EPOfix AND cutting quality sectioning for TEM or Transmission electron microscopy } reveals 241 results.

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

Von: rongchigram79-at-yahoo.com.sg [mailto:rongchigram79-at-yahoo.com.sg]
Gesendet: Montag, 14. Juli 2014 17:54
An: Muß Wolfgang
Betreff: [Microscopy] Experience with Epo-Fix Epoxy as Embedding Medium for Ultramicrotome
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Dear All, recently, i am trying to help a user to section her sample embedded in Epo-Fix epoxy.
As i am not very experienced with ultramicrotome, I decided to section the Epo-Fix epoxy without any sample in it with a glass knife as a start.

Unfortunately, i am unable to section it to a thickness of about 100nm.
What i observed is that the glass knife is only able to section at alternating interval.
I learn that this can be due to either the epoxy is quite soft of [or] the glass knife is not sharp enough (or the block face might still be too big?).

The student told me that that glass transition of the epoxy is above room temperature so there shouldn't be a need to adopt cryo-ultramicrotomy approach. I also attempt to trim the block face to a small size e. g to about 1mm (or slightly less) but still the same alternate sectioning observation is made. I have tried sectioning with biological sample embedded in araldite before and there is no problem achieving thickness of 100nm or less.

My glass knife cutting angle is at 6 degree.

May i ask if i have missed out anything?
Is Epo-fix suitable to be sectioned with a glass knife to achieve a thickness of 100nm?
On my second question, does the Tg of the sample determine if one should use a room-temperature ultramicrotome or a cryo-ultramicrotome?

Cheers,
Yee Yan, Tay
FACTS Lab
Nanyang Technological University

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Hi, All-

If you have old equipment to dump, consider offering it up on this forum
or others. When I posted that I was taking apart an EM lab because the
director had passed away, I got 30 emails from this group! People were
willing to take whole instruments or any number of parts. There are many
who are trying to keep old equipment going, some who can breathe new life
into parts, and now several hacker spaces where kids can be exposed to all
kinds of technology and can learn how to make stuff work.

If you requsted parts from me, I hope I sent you a personal email letting
you know if you "won" something or not. If not, please let me apologize; I
got swamped.

I found homes for a sputter coater, critical point dryer, ultramicrotome,
and plasma asher, all mostly working and fairly easily repaired (I hope).
Even parts from the most desperately old piece of equipment, an old Denton
vacuum evaporator, will have a new life. And the turbo pump and
controller, and cards and valves and gauges and manuals from the SEM have
been sent to several people. I had multiple requests for each item!

So don't forget there are a lot of people out there who could benefit from
pieces and parts. Nerds and hackers keep it going. And now I've managed to
train/corrupt a sweet young undergrad who helped me strip everything, and
she thinks it's some of the most fun she's ever had. Plus we have pictures
of her dropping a turbo with which to torment her four brothers who only
know how to work on mundane cars.

Reuse, recycle, and spread the knowledge and talent.

Aloha,
Tina

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

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Sorry, I made an error : to decelerate the incident beam, the sample is
negatively biased, and not positively as I wrote. Shure, you have all
automatically corrected my miswriting ! ;-)

Regards

Jacques

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Hi

We recently observed some polymer aerogels in un test run for the
purchase of a new SEM, and with recent SEMs it is possible to do nice
things at very low voltage (300 V) in deceleration mode (the sample is
positively biased to slow down the incident electrons, and the SE and BE
are re-accelerated up to the detector), without any coating. It works
well, with imaging up to more than 100 kx, but need some skill to find
the right conditions.

For the cross-section, why not try the ion beam cross-polisher. You
could obtain some test run asking one of the manufacturer (Gatan, Jeol,
Hitachi, Leica Technor-Linda, etc). No embeding, possibily no metalisation.

Hope it helps !

Jacques

Le 14/07/2014 08:31, Ferruccio.Bolla-at-empa.ch a écrit :
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} Dear all,
}
} I am studying cellulose aerogel made from Nanocellulose Fibres (10-30nm diameter and several micron in length).
}
} MATERIAL:
} The material show highly porosity (up to 90% air) and a very intricate structure with meso and nano pores.
}
} SAMPLE:
} Samples are produced via supercritical CO2 drying after filtration and solvent exchange. The resulting sample is a ""paper-like"" fluffy material, 3 cm in diameter and a thickness of 300 micron.
}
} SEM:
} To study the microstructure I tried to embed a small piece of aerogel in araldite. The cross-section surface of the sample was exposed with a diamond microtome and sputtered with 20nm of gold.
}
} In the obtained images is difficult to distinguish between the araldite matrix and the sample (a small change in contrast at low magnification). Worst is that on the surface I don't see any micro-structure. Only a light grey smooth surface.
}
} QUESTION:
} - Is it possible in your opinion to obtain a nice cross section image of my aerogel?
} - I am thinking to change the embedding material with a more stiff epoxy. It could help?
} - Do you have any others ideas?
}
} Many thanks in advance,
} Ferruccio
}
}
} PS: This is my first message. Hope I did everything correct.
}
}
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--

J. Faerber
IPCMS-DSI
Institut de Physique et Chimie des Matériaux de Strasbourg
Département Surfaces et Interfaces
23, rue de Loess ; BP43
67034 Strasbourg CEDEX 2
France

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

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J. Faerber
IPCMS-DSI
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Département Surfaces et Interfaces
23, rue de Loess ; BP43
67034 Strasbourg CEDEX 2
France

Tel 00 33(0)3 88 10 71 01
Fax 00 33(0)3 88 10 72 48
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Dear Yee Yan,

thank you for the information.
IF there is NO mechanical problem with your Leica UCT (stepper motor, feeding) then I am pretty convinced you should be able to cut at least down to 50nm (if not 30nm).
Have you considered also the room climate as a source of failure(i.e. humidity, [cold]draught)? Perhaps some shielding around the block-face - section area could be of benefit.
There exists a description of errors and trouble checking for cutting from the late H. Sitte (who was one of the main inventors of the classical Reichert ultramicrotome series OMU-2,OMU-3, followed by Ultracut, Ultracut E and then UCT etc.).
I shall send this pdf to you, so you perhaps can find out other sources of failure in cutting serial sections with equal thickness (specified therein esp. point 4.: "irregular section thickness").

Hope there will be any other considerations from the list....
Best wishes and good luck,
sincerely,
Wolfgang

} -----Ursprüngliche Nachricht-----
} Von: rongchigram79-at-yahoo.com.sg [mailto:rongchigram79-at-yahoo.com.sg]
} Gesendet: Dienstag, 15. Juli 2014 18:54
} An: Muß Wolfgang
} Betreff: [Microscopy] Re: Re: Experience with Epo-Fix Epoxy as Embedding
} Medium
}
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} Dr Muss, sorry to add in the information. i am using Leica UCT. I use a
} mechanical feed of 100nm. At 300nm, it seems not a problem.
} Cheers,
} Yee Yan
} On Tuesday, 15 July 2014, 1:16, "W.Muss-at-salk.at" {W.Muss-at-salk.at} wrote:
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} Dear Yee Yan Tay,
} IMHO we need also to know which ultramicrotome (thermal or mechanical feed)
} you are using for this approach.
} Epo-Fix is known to me as a specimen mountant(mounting medium) in Materials
} Sciences
} (cf. e.g.
} http://www.struers.com/resources/elements/12/255648/Cold%20Mounting%20table_Ep
} oxies.pdf )
} and unfortunately I haven't experience with sectioning Epo-Fix-Epoxy embedded
} specimens. *)
} It might be this resin too soft or too hard. It might be also the type of
} glass strips you use and how the quality of your knives really is.
} Trials to overcome the problem with your glass knives (including the existing
} angle to be 35-45-55 degr. comparable with the diamond knives 35/45/55
} degr.**)) could be:
} i) sectioning at a (narrower) cutting angle of say 4.5 - 5.5 degree (instead
} of using 6 degr.),
} ii) lowering level of water in the knife boat
} iii) playing around with section speed (ensuring that the section phase is set
} sufficiently long enough, i. e. start cutting at least 1 mm above block edge,
} end of cutting phase as short as possible below /after having sectioned
} through the block at its lower edge).
} iv) trying to harden the block further (oven over night, 65-80 degr.C.)
} v) last but not least - only if available - use of a diamond knife.
}
} With the usual resins for TEM (e.g. Epon 812, Embed812, Glycidether 100
} substitute (for Epon 812), LX-112 etc in former times (also using glass
} knives) - when mixed thoroughly according to the recommandations of LUFT
} (1961) and others I had no problems cutting also bigger specs at least up to
} 4x4 mm (for my thesis I had a really soft resin mixture for cutting whole rat
} hypothalami - 1day PN to 320 days of age - including the ventricle).
}
} Would like to ask you honestly to keep us informed about your problem and
} eventually the solution you found (or even personal Re's you receive on this
} unfortunate but interesting matter),
} best of luck,
} regards
} Wolfgang
}
}
} Wolfgang MUSS PhD
} EM-Lab, Pathology
} SALK-LKH & PMU Salzburg
} SALZBURG, Austria
} [if you need the pdf and can't download, let me know and I will send it to you
} -at- (rongchigram79-at-yahoo.com.sg)]
}
}
} *) From: EPOFIX AND VACUUM: AN EASY METHOD TO MAKE CASTS OF HARD SUBSTRATES
} Jan Kresten Nielsen and Jesper Maiboe
} [Nielsen, Jan Kresten and Maiboe, Jesper 2000. Epofix and Vacuum: An Easy
} Method to Make Casts of Hard Substrates.
} Palaeontologia Electronica, vol. 3, issue 1, art. 2: 10pp., 1.3MB.
} http://palaeo-electronica.org/2000_1/epofix/issue1_00.htm
} Copyright: Palaeontological Association, 15 April 2000
} Submission: 20 August 1999, Acceptance: 24 February 2000]
} see: Palaeontologia Electronica-http://www-odp.tamu.edu/paleo
} "The sectioning properties of Epofix mixture are good. A comparison with other
} embedding media is beyond the scope of this paper. The reader is referred to
} Bromage (1985), who provided a systematic evaluating procedure for casts and
} moulds."
} (Bromage, T. G. 1985. Systematic inquiry in tests of negative/positive replica
} combinations for SEM. Journal of Microscopy, 137:209-216)
}
} **) cf.
} http://cime.epfl.ch/files/content/sites/cime2/files/shared/Files/Teaching/MSE_
} 603_2013_autumn/Chapter%2012%20Sample%20prep%202013.pdf
} (pdf of ppt: TEM Samples preparation_D.Laub_2013 ) scroll down to foil 75:
} Resin:     Knive angle Compression
} Epofix     45°    11%
}         35°    6%
} the further (following foils) 76ff perhaps also are interesting....Foil 80:
} Section thickness 40 - 50nm; Sectioning speed 0.2mm/sec
}
} Searching GOOGLE for { EPOfix AND cutting quality sectioning for TEM or
} Transmission electron microscopy } reveals 241 results.
}
} ========================
}
} Von: rongchigram79-at-yahoo.com.sg [mailto:rongchigram79-at-yahoo.com.sg]
} Gesendet: Montag, 14. Juli 2014 17:54
} An: Muß Wolfgang
} Betreff: [Microscopy] Experience with Epo-Fix Epoxy as Embedding Medium for
} Ultramicrotome
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} Dear All, recently, i am trying to help a user to section her sample embedded
} in Epo-Fix epoxy.
} As i am not very experienced with ultramicrotome, I decided to section the
} Epo-Fix epoxy without any sample in it with a glass knife as a start.
}
} Unfortunately, i am unable to section it to a thickness of about 100nm.
} What i observed is that the glass knife is only able to section at alternating
} interval.
} I learn that this can be due to either the epoxy is quite soft of [or] the
} glass knife is not sharp enough (or the block face might still be too big?).
}
} The student told me that that glass transition of the epoxy is above room
} temperature so there shouldn't be a need to adopt cryo-ultramicrotomy
} approach. I also attempt to trim the block face to a small size e. g to about
} 1mm (or slightly less) but still the same alternate sectioning observation is
} made. I have tried sectioning with biological sample embedded in araldite
} before and there is no problem achieving thickness of 100nm or less.
}
} My glass knife cutting angle is at 6 degree.
}
}
} May i ask if i have missed out anything?
} Is Epo-fix suitable to be sectioned with a glass knife to achieve a thickness
} of 100nm?
} On my second question, does the Tg of the sample determine if one should use a
} room-temperature ultramicrotome or a cryo-ultramicrotome?
}
} Cheers,
} Yee Yan, Tay
} FACTS Lab
} Nanyang Technological University
}
}
}
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} 7, 34 -- Reply-To: YY YY {rongchigram79-at-yahoo.com.sg}
} 7, 34 -- Subject: Experience with Epo-Fix Epoxy as Embedding Medium for
} Ultramicrotome
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} 23, 40 -- Subject: [Microscopy] Re: Experience with Epo-Fix Epoxy as Embedding
} Medium
} 23, 40 -- for Ultramicrotome
} 23, 40 -- Thread-Topic: [Microscopy] Re: Experience with Epo-Fix Epoxy as
} Embedding
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} 36, 53 -- Subject: Re: [Microscopy] Re: Experience with Epo-Fix Epoxy as
} Embedding Medium
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8, 40 -- Subject: [Microscopy] Re: Re: Experience with Epo-Fix Epoxy as Embedding
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Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Dear all,
in my reply below I wrote " "late H. Sitte". I apologize for this my "semantic" fault, since I meant "retired Prof. H. Sitte"

John Minter sent me an e-mail regarding this untrue fact and so I cann tell:

Prof. Helmuth Sitte has celebrated his 85th birthday on 6th of May 2103.
His company still is protocolled (perhaps also his living town and address) at
Dr. Hellmuth Sitte
Reitherspitzstraße 166
A-6100 Seefeld
Austria

I confused here the death of his brother I think as well as the closure of the famous courses on Preparation techniques in (T)EM.
So, I guess this is really a big mistake I have to correct via MSA-Listserver

Regards
Wolfgang

Von: W.Muss-at-salk.at [mailto:W.Muss-at-salk.at]
Gesendet: Mittwoch, 16. Juli 2014 09:44
An: Muß Wolfgang
Betreff: CTRL_[Microscopy] Re to MSA-Listserver: Re: Experience with Epo-Fix Epoxy as Embedding

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Dear Yee Yan,

thank you for the information.
IF there is NO mechanical problem with your Leica UCT (stepper motor, feeding) then I am pretty convinced you should be able to cut at least down to 50nm (if not 30nm).
Have you considered also the room climate as a source of failure (i.e. humidity, [cold]draught)? Perhaps some shielding around the block-face - section area could be of benefit.
There exists a description of errors and trouble checking for cutting from the late H. Sitte (who was one of the main inventors of the classical Reichert ultramicrotome series OMU-2, OMU-3, followed by Ultracut, Ultracut E and then UCT etc.).
I shall send this pdf to you, so you perhaps can find out other sources of failure in cutting serial sections with equal thickness (specified therein esp. point 4.: "irregular section thickness").

Hope there will be any other considerations from the list....
Best wishes and good luck,
sincerely,
Wolfgang

} -----Ursprüngliche Nachricht-----
} Von: rongchigram79-at-yahoo.com.sg [mailto:rongchigram79-at-yahoo.com.sg]
} Gesendet: Dienstag, 15. Juli 2014 18:54
} An: Muß Wolfgang
} Betreff: [Microscopy] Re: Re: Experience with Epo-Fix Epoxy as Embedding
} Medium
}
} ----------------------------------------------------------------------------
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}
} Dr Muss, sorry to add in the information. i am using Leica UCT. I use a
} mechanical feed of 100nm. At 300nm, it seems not a problem.
} Cheers,
} Yee Yan
} On Tuesday, 15 July 2014, 1:16, "W.Muss-at-salk.at" {W.Muss-at-salk.at} wrote:
} ----------------------------------------------------------------------------
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} Dear Yee Yan Tay,
} IMHO we need also to know which ultramicrotome (thermal or mechanical feed)
} you are using for this approach.
} Epo-Fix is known to me as a specimen mountant(mounting medium) in Materials
} Sciences
} (cf. e.g.
} http://www.struers.com/resources/elements/12/255648/Cold%20Mounting%20table_Ep
} oxies.pdf )
} and unfortunately I haven't experience with sectioning Epo-Fix-Epoxy embedded
} specimens. *)
} It might be this resin too soft or too hard. It might be also the type of
} glass strips you use and how the quality of your knives really is.
} Trials to overcome the problem with your glass knives (including the existing
} angle to be 35-45-55 degr. comparable with the diamond knives 35/45/55
} degr.**)) could be:
} i) sectioning at a (narrower) cutting angle of say 4.5 - 5.5 degree (instead
} of using 6 degr.),
} ii) lowering level of water in the knife boat
} iii) playing around with section speed (ensuring that the section phase is set
} sufficiently long enough, i. e. start cutting at least 1 mm above block edge,
} end of cutting phase as short as possible below /after having sectioned
} through the block at its lower edge).
} iv) trying to harden the block further (oven over night, 65-80 degr.C.)
} v) last but not least - only if available - use of a diamond knife.
}
} With the usual resins for TEM (e.g. Epon 812, Embed812, Glycidether 100
} substitute (for Epon 812), LX-112 etc in former times (also using glass
} knives) - when mixed thoroughly according to the recommandations of LUFT
} (1961) and others I had no problems cutting also bigger specs at least up to
} 4x4 mm (for my thesis I had a really soft resin mixture for cutting whole rat
} hypothalami - 1day PN to 320 days of age - including the ventricle).
}
} Would like to ask you honestly to keep us informed about your problem and
} eventually the solution you found (or even personal Re's you receive on this
} unfortunate but interesting matter),
} best of luck,
} regards
} Wolfgang
}
}
} Wolfgang MUSS PhD
} EM-Lab, Pathology
} SALK-LKH & PMU Salzburg
} SALZBURG, Austria
} [if you need the pdf and can't download, let me know and I will send it to you
} -at- (rongchigram79-at-yahoo.com.sg)]
}
}
} *) From: EPOFIX AND VACUUM: AN EASY METHOD TO MAKE CASTS OF HARD SUBSTRATES
} Jan Kresten Nielsen and Jesper Maiboe
} [Nielsen, Jan Kresten and Maiboe, Jesper 2000. Epofix and Vacuum: An Easy
} Method to Make Casts of Hard Substrates.
} Palaeontologia Electronica, vol. 3, issue 1, art. 2: 10pp., 1.3MB.
} http://palaeo-electronica.org/2000_1/epofix/issue1_00.htm
} Copyright: Palaeontological Association, 15 April 2000
} Submission: 20 August 1999, Acceptance: 24 February 2000]
} see: Palaeontologia Electronica-http://www-odp.tamu.edu/paleo
} "The sectioning properties of Epofix mixture are good. A comparison with other
} embedding media is beyond the scope of this paper. The reader is referred to
} Bromage (1985), who provided a systematic evaluating procedure for casts and
} moulds."
} (Bromage, T. G. 1985. Systematic inquiry in tests of negative/positive replica
} combinations for SEM. Journal of Microscopy, 137:209-216)
}
} **) cf.
} http://cime.epfl.ch/files/content/sites/cime2/files/shared/Files/Teaching/MSE_
} 603_2013_autumn/Chapter%2012%20Sample%20prep%202013.pdf
} (pdf of ppt: TEM Samples preparation_D.Laub_2013 ) scroll down to foil 75:
} Resin:     Knive angle Compression
} Epofix     45°    11%
}         35°    6%
} the further (following foils) 76ff perhaps also are interesting....Foil 80:
} Section thickness 40 - 50nm; Sectioning speed 0.2mm/sec
}
} Searching GOOGLE for { EPOfix AND cutting quality sectioning for TEM or
} Transmission electron microscopy } reveals 241 results.
}
} ========================
}
} Von: rongchigram79-at-yahoo.com.sg [mailto:rongchigram79-at-yahoo.com.sg]
} Gesendet: Montag, 14. Juli 2014 17:54
} An: Muß Wolfgang
} Betreff: [Microscopy] Experience with Epo-Fix Epoxy as Embedding Medium for
} Ultramicrotome
} ---------------------------------------------------------------------------
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} Dear All, recently, i am trying to help a user to section her sample embedded
} in Epo-Fix epoxy.
} As i am not very experienced with ultramicrotome, I decided to section the
} Epo-Fix epoxy without any sample in it with a glass knife as a start.
}
} Unfortunately, i am unable to section it to a thickness of about 100nm.
} What i observed is that the glass knife is only able to section at alternating
} interval.
} I learn that this can be due to either the epoxy is quite soft of [or] the
} glass knife is not sharp enough (or the block face might still be too big?).
}
} The student told me that that glass transition of the epoxy is above room
} temperature so there shouldn't be a need to adopt cryo-ultramicrotomy
} approach. I also attempt to trim the block face to a small size e. g to about
} 1mm (or slightly less) but still the same alternate sectioning observation is
} made. I have tried sectioning with biological sample embedded in araldite
} before and there is no problem achieving thickness of 100nm or less.
}
} My glass knife cutting angle is at 6 degree.
}
}
} May i ask if i have missed out anything?
} Is Epo-fix suitable to be sectioned with a glass knife to achieve a thickness
} of 100nm?
} On my second question, does the Tg of the sample determine if one should use a
} room-temperature ultramicrotome or a cryo-ultramicrotome?
}
} Cheers,
} Yee Yan, Tay
} FACTS Lab
} Nanyang Technological University
}
}
}
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Email: iskandar-at-gfe.rwth-aachen.de
Name: Mohamad Riza Iskandar

Organization: Central Facility for Electron Microscopy RWTH Aachen University

Title-Subject: [Filtered] Running Digital Micrograph on Mac OSX

Message: Dear all,

I'm trying to run Digital Micrograph on my Mac (10.9.4) but unfortunately I get problem with the
installation process. I've followed the direction as written in one article in Microscopy today
(March 2012) but I always get problem when installing the Digital Micrograph. The installation
process always stop with an error message of "Removing device DMVIDEO, cleanup FAILURE". I did
install everything I knew as prerequisite as mentioned in that article, but still no luck.

I would appreciate if someone can help me to solve this problem. Any suggestions via private message
are also welcome.

Thank you in advanced,

Best regards,

Riza Iskandar

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Dear Prof Muss, thank you very much for your kindness in providing all the valuable advices and experiences! Let me read them slowly and try on my machine. I also have a Leica UC7 to try on. My country is hot and humid but the lab is kept at 40-50% humidity. I am not sure if it will contribute to the degrading of the Epo-fix.

Thank you once again!!

Cheers,
Yee Yan

On Wednesday, July 16, 2014 3:59 PM, "W.Muss-at-salk.at" {W.Muss-at-salk.at} wrote:

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Dear Yee Yan,

thank you for the information.
IF there is NO mechanical problem with your Leica UCT (stepper motor, feeding) then I am pretty convinced you should be able to cut at least down to 50nm (if not 30nm).
Have you considered also the room climate as a source of failure(i.e. humidity, [cold]draught)? Perhaps some shielding around the block-face - section area could be of benefit.
There exists a description of errors and trouble checking for cutting from the late H. Sitte (who was one of the main inventors of the classical Reichert ultramicrotome series OMU-2,OMU-3, followed by Ultracut, Ultracut E and then UCT etc.).
I shall send this pdf to you, so you perhaps can find out other sources of failure in cutting serial sections with equal thickness (specified therein esp. point 4.: "irregular section thickness").

Hope there will be any other considerations from the list....
Best wishes and good luck,
sincerely,
Wolfgang

} -----Ursprüngliche Nachricht-----
} Von: rongchigram79-at-yahoo.com.sg [mailto:rongchigram79-at-yahoo.com.sg]
} Gesendet: Dienstag, 15. Juli 2014 18:54
} An: Muß Wolfgang
} Betreff: [Microscopy] Re: Re: Experience with Epo-Fix Epoxy as Embedding
} Medium
}
} ----------------------------------------------------------------------------
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}
} Dr Muss, sorry to add in the information. i am using Leica UCT. I use a
} mechanical feed of 100nm. At 300nm, it seems not a problem.
} Cheers,
} Yee Yan
} On Tuesday, 15 July 2014, 1:16, "W.Muss-at-salk.at" {W.Muss-at-salk.at} wrote:
} ----------------------------------------------------------------------------
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} Dear Yee Yan Tay,
} IMHO we need also to know which ultramicrotome (thermal or mechanical feed)
} you are using for this approach.
} Epo-Fix is known to me as a specimen mountant(mounting medium) in Materials
} Sciences
} (cf. e.g.
} http://www.struers.com/resources/elements/12/255648/Cold%20Mounting%20table_Ep
} oxies.pdf )
} and unfortunately I haven't experience with sectioning Epo-Fix-Epoxy embedded
} specimens. *)
} It might be this resin too soft or too hard. It might be also the type of
} glass strips you use and how the quality of your knives really is.
} Trials to overcome the problem with your glass knives (including the existing
} angle to be 35-45-55 degr. comparable with the diamond knives 35/45/55
} degr.**)) could be:
} i) sectioning at a (narrower) cutting angle of say 4.5 - 5.5 degree (instead
} of using 6 degr.),
} ii) lowering level of water in the knife boat
} iii) playing around with section speed (ensuring that the section phase is set
} sufficiently long enough, i. e. start cutting at least 1 mm above block edge,
} end of cutting phase as short as possible below /after having sectioned
} through the block at its lower edge).
} iv) trying to harden the block further (oven over night, 65-80 degr.C.)
} v) last but not least - only if available - use of a diamond knife.
}
} With the usual resins for TEM (e.g. Epon 812, Embed812, Glycidether 100
} substitute (for Epon 812), LX-112 etc in former times (also using glass
} knives) - when mixed thoroughly according to the recommandations of LUFT
} (1961) and others I had no problems cutting also bigger specs at least up to
} 4x4 mm (for my thesis I had a really soft resin mixture for cutting whole rat
} hypothalami - 1day PN to 320 days of age - including the ventricle).
}
} Would like to ask you honestly to keep us informed about your problem and
} eventually the solution you found (or even personal Re's you receive on this
} unfortunate but interesting matter),
} best of luck,
} regards
} Wolfgang
}
}
} Wolfgang MUSS PhD
} EM-Lab, Pathology
} SALK-LKH & PMU Salzburg
} SALZBURG, Austria
} [if you need the pdf and can't download, let me know and I will send it to you
} -at- (rongchigram79-at-yahoo.com.sg)]
}
}
} *) From: EPOFIX AND VACUUM: AN EASY METHOD TO MAKE CASTS OF HARD SUBSTRATES
} Jan Kresten Nielsen and Jesper Maiboe
} [Nielsen, Jan Kresten and Maiboe, Jesper 2000. Epofix and Vacuum: An Easy
} Method to Make Casts of Hard Substrates.
} Palaeontologia Electronica, vol. 3, issue 1, art. 2: 10pp., 1.3MB.
} http://palaeo-electronica.org/2000_1/epofix/issue1_00.htm
} Copyright: Palaeontological Association, 15 April 2000
} Submission: 20 August 1999, Acceptance: 24 February 2000]
} see: Palaeontologia Electronica-http://www-odp.tamu.edu/paleo
} "The sectioning properties of Epofix mixture are good. A comparison with other
} embedding media is beyond the scope of this paper. The reader is referred to
} Bromage (1985), who provided a systematic evaluating procedure for casts and
} moulds."
} (Bromage, T. G. 1985. Systematic inquiry in tests of negative/positive replica
} combinations for SEM. Journal of Microscopy, 137:209-216)
}
} **) cf.
} http://cime.epfl.ch/files/content/sites/cime2/files/shared/Files/Teaching/MSE_
} 603_2013_autumn/Chapter%2012%20Sample%20prep%202013.pdf
} (pdf of ppt: TEM Samples preparation_D.Laub_2013 ) scroll down to foil 75:
} Resin:     Knive angle Compression
} Epofix     45°    11%
}         35°    6%
} the further (following foils) 76ff perhaps also are interesting....Foil 80:
} Section thickness 40 - 50nm; Sectioning speed 0.2mm/sec
}
} Searching GOOGLE for { EPOfix AND cutting quality sectioning for TEM or
} Transmission electron microscopy } reveals 241 results.
}
} ========================
}
} Von: rongchigram79-at-yahoo.com.sg [mailto:rongchigram79-at-yahoo.com.sg]
} Gesendet: Montag, 14. Juli 2014 17:54
} An: Muß Wolfgang
} Betreff: [Microscopy] Experience with Epo-Fix Epoxy as Embedding Medium for
} Ultramicrotome
} ---------------------------------------------------------------------------
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} ----------------------------------------------------------------------------
}
} Dear All, recently, i am trying to help a user to section her sample embedded
} in Epo-Fix epoxy.
} As i am not very experienced with ultramicrotome, I decided to section the
} Epo-Fix epoxy without any sample in it with a glass knife as a start.
}
} Unfortunately, i am unable to section it to a thickness of about 100nm.
} What i observed is that the glass knife is only able to section at alternating
} interval.
} I learn that this can be due to either the epoxy is quite soft of [or] the
} glass knife is not sharp enough (or the block face might still be too big?).
}
} The student told me that that glass transition of the epoxy is above room
} temperature so there shouldn't be a need to adopt cryo-ultramicrotomy
} approach. I also attempt to trim the block face to a small size e. g to about
} 1mm (or slightly less) but still the same alternate sectioning observation is
} made. I have tried sectioning with biological sample embedded in araldite
} before and there is no problem achieving thickness of 100nm or less.
}
} My glass knife cutting angle is at 6 degree.
}
}
} May i ask if i have missed out anything?
} Is Epo-fix suitable to be sectioned with a glass knife to achieve a thickness
} of 100nm?
} On my second question, does the Tg of the sample determine if one should use a
} room-temperature ultramicrotome or a cryo-ultramicrotome?
}
} Cheers,
} Yee Yan, Tay
} FACTS Lab
} Nanyang Technological University
}
}
}
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} 23, 40 -- Subject: [Microscopy] Re: Experience with Epo-Fix Epoxy as Embedding
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} 36, 53 -- Subject: Re: [Microscopy] Re: Experience with Epo-Fix Epoxy as
} Embedding Medium
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8, 40 -- Subject: [Microscopy] Re: Re: Experience with Epo-Fix Epoxy as Embedding
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From pharmacy-express1-at-acbrecovery.com Wed Jul 16 22:49:24 2014
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We are having internal discussions about the structure and support for our microscopy facility and I would like to take an informal poll of others who run departmental or campus microscopy facilities.

1. Does your institution support in part or fully, a facility scientist(s) to help run the facility day to day
2. Do you receive some form of compensation as the person in charge of the facility in terms of salary or reduced teaching or other?
3. Does your institution help support repairs or service contracts for instruments?

Will compile results and post if desired. Any extra information appreciated.
Thanks- Dave

Dr. David Knecht
Professor of Molecular and Cell Biology
Core Microscopy Facility Director
University of Connecticut
Storrs, CT 06269
860-486-2200

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All,

We have many fume hoods in the department but no perchloric hood. I'm
looking for an electro polisher recipe for making TEM foils of
nickel-chrome-aluminum based superalloys that doesn't require perchloric
acid.

I appreciate any help you can lend.

Cheers Owen

--
Owen P Mills
Michigan Technological University
1400 Townsend Dr
Houghton, MI 49931
906-369-1875

==============================Original Headers==============================
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Dear colleagues,

The Advanced Imaging Center (AIC) at HHMI Janelia Research Campus is
jointly sponsored by the Howard Hughes Medical Institute and the Gordon
and Betty Moore Foundation. Our mission is to make cutting-edge,
pre-commercial microscopes developed at Janelia available to visiting
scientists, maximizing the impact of the latest developments in optical
instruments and emerging microscopy technologies.

We encourage applications from scientists who are addressing significant
scientific questions that require measurements of cellular/molecular
behavior at spatial and/or temporal resolutions that would only be
possible for them through access to the AIC.

Under the Program, visiting scientists will spend 1-2 weeks (or longer if
there is justified scientific reason) at Janelia to conduct experiments
on the scope with the support of the AIC team.

The currently available microscopes, which provide advanced capabilities
that are not yet widely available, are:

- Interferometric photoactivated localization microscope (iPALM)
- Single-molecule total internal reflection fluorescence (smTIRF)
microscope
- Aberration-corrected multifocus microscope (acMFM)
- Lattice light sheet microscope (2nd generation bessel beam microscope)
- Live cell multicolor structured illumination microscope (SIM)

Our website, http://www.janelia.org/aic provides more details about these
unique instruments.

Our first Call-for-Proposals begins today, July 15, 2014, and closes on
August 15, 2014. Proposals will be evaluated based on scientific merit
through a two-tier review process by AIC/Janelia scientists and by a
review panel of scientists from HHMI/Janelia, GBMF, and extramural
imaging experts. Additional information and instructions on how apply
are available here: http://www.janelia.org/aic

Scientists whose proposals are selected will travel to Janelia to use the
facility. The Janelia Visitor Program will cover the cost of lodging for
the visiting scientist, experiments, technical support from our
applications scientists, and scope time.

Email aic-at-janelia.hhmi.org or me with direct inquiries.

Please help us by spreading the word to your colleagues. Here is the
link to the HHMI press release. http://tinyurl.com/ncbwwbp

Regards,

Teng-Leong Chew, PhD
Director, Advanced Imaging Center
Howard Hughes Medical Institute at Janelia
Ashburn, VA 20147
(571) 209-4676
chewt-at-janelia.hhmi.org

==============================Original Headers==============================
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From drugs-canadian10-at-hinet.net Thu Jul 17 13:52:58 2014
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Email: gary-at-gaugler.com
Name: Dr Gary Gaugler

Organization: Microtechnics, Inc.

Title-Subject: [Filtered] SFEG SEMs

Message: Hello Listers:

I am looking for a used SFEG and know some makes and models but could use some help to round out the
search list. First, did LEO make a SFEG before the Zeiss Supra40 and if so, what model is it?
Second, are there any small/medium chamber size SFEG SEMs like the Supra40 chamber from other
makers? The largest chamber system I had was the Zeiss Supra55. Nice but not that big of chamber
needed. I'm doing pin stubs and metallurgical coupons. High resolution, EDS and mag up to 300KX with
dual SE detectors and BSD are about it. I think that Hitachi only does CFE but JEOL does CFE and
TFE. I'd like to get a system up and running by the end of this year or early next year depending on
what I can find with PC control and is affordable. All inputs are appreciated.

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From affordable-drugstore10-at-jeffco.k12.co.us Thu Jul 17 23:00:48 2014
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Dear listers,

I'd like to have your opinion about the use or non-use of sodium borohydride in the procedure for preparing cells or tissue for immunology. The fixative that has been used is 4% paraformaldehyde and 0.1% glutaraldehyde in phosphate buffer which give us better membranes rather than not using any glutaraldehyde.

I know that the sodium borohydride changes the unbound aldehyde to an alcohol but how much aldehyde can be left in the cells after all the washes?

Opinions and experiences related are those of Pat Connelly and do not represent the NIH. This message is not confidential and can be freely shared and reproduced.
Pat
Patricia Stranen Connelly
Research Assistant
NHLBI EM Core Facility
National Institutes of Health
Building 14E Room 111B
Bethesda, MD 20892-5570
301-496-3491
connellyps-at-mail.nih.gov
http://www.nhlbi.nih.gov/research/intramural/researchers/core/electron-microscopy-core/index.html

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I am looking to purchase a new dimpling grinder, I've only used the Gatan
model but am curious about the VCR and Fischione products. If you have any
comments on the tools or comparison between manufacturers could you please
contact me off list?

Thank you,
Leslie Thompson

Staff Scientist/Engineer
IBM Research-Almaden

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Dear listers

I recently got back from MMC 2014 in the UK (microscopy convention)
and I was truly surprised that all the SEM manufactures still do not
allow users to calibrate the scale bar on the new SEMS. The responses
were all the same "our product IS calibrated in the factory, there is
no need to mess with it" or "if you want to calibrate then why don't
you run a standard in parallel?". When I push the reps most confess
that the calibration is done at best on a ~2000l/mm grating which on
our SEM does not equal good high mag measurements 400kX+. I am also
amazed that most companies also do not correct for slow scan drift in
the pixel so seeing as there is never a vertical scale bar in the
images (well not ours) how can you believe the measurements?

I am not sure if it just me or my experience with older SEMs but I am
sure there are many fields where a user might want to have independent
calibration? I know this has been much discussed in the past but with
digital recording we seem to have lost respect for calibration
(perhaps this is just my jaded view?)

In our center we use a third party imaging system which is calibrated
by ourselves against standards of our own making and verified with
commercial standards and we get very good accuracy and reproducibility
(we think). We account for all changes in beam and vacuum state. It
certainly agrees with the TEM, XRD and other techniques we might use.
The same can not be said for trusting the machine scalebars.We have
very old machines. I am right in not trusting new machines as much as
the old ones?

I would be interested to hear what peoples experiences are with this
mag calibration issue especially in high mag. Has anyone found a way
of doing this within a modern SEM? What standards do you use? Do you
think the standards are good enough?

I also wanted to see if anyone would be interested in a little global
test. We make LED devices in house so measurement for us is critical.
I would like to propose that I send out to anyone interested a small
piece of a bulk wafer (the same one!) with a pin array on it with a
set size, shape, spacing. The aim would be to take pictures at set
mags eg 10, 100, 250, 500kx and then make measurements based on your
own lab method. You could then return the data to me along with the
make and model of the SEM and your method of imaging/calibration. I
would then collate and circulate the anonymised data to the list. This
is not about the age of the SEM, the max mag, the type of source or
even the lab budget I am just really interested in seeing what the
real variation is out there.

If you have an interest in this and perhaps would like to participate
in my little test (I would welcome instrument manufacturers to take
part). If you want to reply to me on or off list any responses,
opinions will be kept anonymous if requested.

Thanks for reading
John

==============================Original Headers==============================
8, 26 -- From john.mitchels-at-gmail.com Sat Jul 19 15:41:08 2014
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8, 26 -- Subject: Mag calibration variation with SEMs, proposed: a little test
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From medications_best9-at-ftninc.com Sun Jul 20 16:50:17 2014
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John:

My mentor at HP, Nancy Phillips always had us check the Mag Cal on our many SEMs every quarter. We used an MRS 5 at that time, NIST traceable. If we found a measurement of the MRS 5 that deviated more than 5% we let FEI know and they (Jeff Cohen) would come change the Calibration for us! It was nice as we had an FEI service Engineer, Jeff, onsite. We really never saw deviations above 5% ever, a testament to Jeff's ability.

Now that I am at Oregon State, I have implemented a similar policy. I purchased an MRS 6, at $5k they are not cheap, and then I check all our SEMs at a random selection of Magnifications, voltages, detectors, ... We then update an excel spreadsheet that is shared between my coworker and I on a google drive. I have not seen deviation above 5%. Again, testament to our FEI guys: Bob Johnson and Frank. Just today I checked our Nova NanoSEM at 5kx, 10kx using the ETD and 100kx with the TLD in immersion. Deviations of the MRS 6 pitch, were well below 5%.

I would certainly participate in an inter lab comparison with your LED. I did one of these with the ANSI TAG group on gold nanoparticles a few years back and it was fun and educational.

Cheers,

Pete Eschbach
Oregon State University Electron Microscope Facility Director
541 737 5645

-----Original Message-----
X-from: john.mitchels-at-gmail.com [mailto:john.mitchels-at-gmail.com]
Sent: Saturday, July 19, 2014 2:13 PM
To: Eschbach, Peter

Dear listers

I recently got back from MMC 2014 in the UK (microscopy convention) and I was truly surprised that all the SEM manufactures still do not allow users to calibrate the scale bar on the new SEMS. The responses were all the same "our product IS calibrated in the factory, there is no need to mess with it" or "if you want to calibrate then why don't you run a standard in parallel?". When I push the reps most confess that the calibration is done at best on a ~2000l/mm grating which on our SEM does not equal good high mag measurements 400kX+. I am also amazed that most companies also do not correct for slow scan drift in the pixel so seeing as there is never a vertical scale bar in the images (well not ours) how can you believe the measurements?

I am not sure if it just me or my experience with older SEMs but I am sure there are many fields where a user might want to have independent calibration? I know this has been much discussed in the past but with digital recording we seem to have lost respect for calibration (perhaps this is just my jaded view?)

In our center we use a third party imaging system which is calibrated by ourselves against standards of our own making and verified with commercial standards and we get very good accuracy and reproducibility (we think). We account for all changes in beam and vacuum state. It certainly agrees with the TEM, XRD and other techniques we might use.
The same can not be said for trusting the machine scalebars.We have very old machines. I am right in not trusting new machines as much as the old ones?

I would be interested to hear what peoples experiences are with this mag calibration issue especially in high mag. Has anyone found a way of doing this within a modern SEM? What standards do you use? Do you think the standards are good enough?

I also wanted to see if anyone would be interested in a little global test. We make LED devices in house so measurement for us is critical.
I would like to propose that I send out to anyone interested a small piece of a bulk wafer (the same one!) with a pin array on it with a set size, shape, spacing. The aim would be to take pictures at set mags eg 10, 100, 250, 500kx and then make measurements based on your own lab method. You could then return the data to me along with the make and model of the SEM and your method of imaging/calibration. I would then collate and circulate the anonymised data to the list. This is not about the age of the SEM, the max mag, the type of source or even the lab budget I am just really interested in seeing what the real variation is out there.

If you have an interest in this and perhaps would like to participate in my little test (I would welcome instrument manufacturers to take part). If you want to reply to me on or off list any responses, opinions will be kept anonymous if requested.

Thanks for reading
John

==============================Original Headers==============================
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8, 26 -- Subject: Mag calibration variation with SEMs, proposed: a little test 8, 26 -- From: John Mitchels {john.mitchels-at-gmail.com} 8, 26 -- To: Microscopy-at-microscopy.com 8, 26 -- Content-Type: text/plain; charset=UTF-8 ==============================End of - Headers==============================

==============================Original Headers==============================
20, 40 -- From Peter.Eschbach-at-oregonstate.edu Mon Jul 21 13:55:45 2014
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20, 40 -- Subject: RE: [Microscopy] Mag calibration variation with SEMs, proposed: a
20, 40 -- little test
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Name: Sandra Keller

Organization: TA

Title-Subject: [Filtered] TEM-Particle counting software

Message: Hi All:
I am looking for recommendations of particle counting programs that function reliably and require
minimal interaction to obtain sizing information on particles in TEM images. If you have an
estimate on the program cost that would also be helpful. Programs I have used in the past, seem to
be fussy and difficult to use, especially when used on particles with limited contrast.
Thanks in advance,
Sandra

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Hello John
Happy to participate to your test. My impression from my Jeol JSM5600LV is
that a variation of at least 5% in magnification scale may occur within two
measurements on the same sample with the same microscope settings, provided
that the microscope was shut down and restarted between these two
measurements. Also, I have an external scan generation device (DISS5 from
"pointelectronic") that is also calibrated and can be tested for accuracy.
With best wishes
yorgos

Dr Yorgos Nikas
Athens Innovative Microscopy
Skra 36 Voula 16673 GREECE

Tel/fax +30 210 8957677
mobile +30 6945 107477
www.eikonika.net www.aim.cat
************************************

----- Original Message -----
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To: {eikonika-at-otenet.gr}
Sent: Saturday, July 19, 2014 11:56 PM

On 22.07.2014 02:34, microscopylistserver-noreply-at-microscopy.com wrote:
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} Email: sk-at-tem-analysis.com
} Name: Sandra Keller
}
} Organization: TA
}
} Title-Subject: [Filtered] TEM-Particle counting software
}
} Message: Hi All:
} I am looking for recommendations of particle counting programs that function reliably and require
} minimal interaction to obtain sizing information on particles in TEM images. If you have an
} estimate on the program cost that would also be helpful. Programs I have used in the past, seem to
} be fussy and difficult to use, especially when used on particles with limited contrast.
} Thanks in advance,
} Sandra
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Hi Sandra, Look up the page http://www.jmicrovision.com/. Maybe this
piece of soft would fulfill your expectations.
All the best

--
Jean-Michel Jaquet, PhD
Geomicrobiology Group
Section of Earth and Environmental Sciences
University of Geneva
13 rue des Maraîchers
CH-1211 Geneva 3
Switzerland
http://cms.unige.ch/sciences/terre/research/Groups/limnogeology/limno.php

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Email: sk-at-tem-analysis.com
Name: Sandra Keller

Organization: TA

Title-Subject: [Filtered] TEM-Particle measurement software-corrected

Message: Sorry, I should entitled my post: TEM-particle measurement software rather than
TEM-Particle counting software. Again, I am looking for recommendations of particle measurement
programs that function reliably and require minimal interaction to obtain sizing information on
particles in TEM images. If you have an estimate on the program cost that would also be helpful.
Programs that I have used in the past, seem to be fussy and difficult to use, especially when used
on particles with limited contrast.
Thanks in advance,
Sandra

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Yorgos,
That can (will) definitely happen if you are not, in some way, normalizing
your hysteresis. Depending upon the microscope, there are different ways to
normalize. Some involve reversing the lens currents for a bit, while others
simply turn off the lens currents. Either way, your lenses can be
considered "normalized" if you use what ever routine is appropriate for your
scope twice in a row and the image doesn't need refocusing at 1000X.

As an experiment, focus at a long working distance and collect an image of
something that you can measure. Run your focus control all the way up
(shortest working distance), then back down to focus and collect another
image. I think you'll find the actual mags may be the same but, with most
instruments, the readouts will be quite different because in the second
image the instrument thinks the WD is much longer, so the WD multiplier is
less than in the first image.

Ken Converse
owner

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

-----Original Message-----
X-from: eikonika-at-otenet.gr [mailto:eikonika-at-otenet.gr]
Sent: Monday, July 21, 2014 10:27 PM
To: kenconverse-at-qualityimages.biz

Hello John
Happy to participate to your test. My impression from my Jeol JSM5600LV is
that a variation of at least 5% in magnification scale may occur within two
measurements on the same sample with the same microscope settings, provided
that the microscope was shut down and restarted between these two
measurements. Also, I have an external scan generation device (DISS5 from
"pointelectronic") that is also calibrated and can be tested for accuracy.
With best wishes
yorgos

Dr Yorgos Nikas
Athens Innovative Microscopy
Skra 36 Voula 16673 GREECE

Tel/fax +30 210 8957677
mobile +30 6945 107477
www.eikonika.net www.aim.cat
************************************

----- Original Message -----
X-from: {john.mitchels-at-gmail.com}
To: {eikonika-at-otenet.gr}
Sent: Saturday, July 19, 2014 11:56 PM

We recently had the Gatan HAADF detector off of our JEOL 2100F. I measured inner and outer diameters and found that the angles in the manual at the specified camera length were not correct. So, in addition to going through the exercise of measuring the collection angles at different camera lengths for my GIF, I carefully measured the camera lengths at the HAADF position and calculated the inner and outer collection angles for the HAADF detector.

I work with a lot of different materials, from polymers and light element ceramics to heavy element alloys. My question is there a method to optimize the cameral length, i.e. collection angles for a particular material system when you know the elements? Should I base it on the EELS characteristic angles for the elements? If I do, use the K shells? If I want to maximize the contrast, should I try to exclude the lower Z characteristic angle from the detector?

-Scott Walck

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Hi Scott,
I did this task using NIST's Elastic32 program. This calculates the
differential electron scattering cross section for elements; combined with a
quick integration over the correct angles, relative counts can be
determined.
Regards,
Larry Scipioni

-----Original Message-----
X-from: s.walck-at-comcast.net [mailto:s.walck-at-comcast.net]
Sent: Thursday, July 24, 2014 9:07 AM
To: LES-at-ZSGENETICS.COM

We recently had the Gatan HAADF detector off of our JEOL 2100F. I measured
inner and outer diameters and found that the angles in the manual at the
specified camera length were not correct. So, in addition to going through
the exercise of measuring the collection angles at different camera lengths
for my GIF, I carefully measured the camera lengths at the HAADF position
and calculated the inner and outer collection angles for the HAADF detector.

I work with a lot of different materials, from polymers and light element
ceramics to heavy element alloys. My question is there a method to optimize
the cameral length, i.e. collection angles for a particular material system
when you know the elements? Should I base it on the EELS characteristic
angles for the elements? If I do, use the K shells? If I want to maximize
the contrast, should I try to exclude the lower Z characteristic angle from
the detector?

-Scott Walck

==============================Original Headers==============================
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Email: tina.williams-at-ars.usda.gov
Name: Tina Williams

Organization: USDA

Title-Subject: [Filtered] 1.CryoJane tape transfer 2.Recommendations for confocal system

Message: Hi,
Two questions:

1. Does anyone know of CryoJane Tape transfer users in the San Francisco Bay Area or nearby, who
would be willing to demonstrate their unit? We are wondering if it will work with our current
cryostat CM3000, and plant tissues?

2. Does anyone have recommendations for models/components on a confocal system for imaging plant
tissues? We are considering purchasing a confocal system.

Thank you,
Tina

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Email: sk-at-tem-analysis.com
Name: Sandra Keller

Organization: TA

Title-Subject: [Filtered] TEM-Particle measurement software-corrected

Message: Sorry, I should entitled my post: TEM-particle measurement software rather than
TEM-Particle counting software. Again, I am looking for recommendations of particle measurement
programs that function reliably and require minimal interaction to obtain sizing information on
particles in TEM images. If you have an estimate on the program cost that would also be helpful.
Programs that I have used in the past, seem to be fussy and difficult to use, especially when used
on particles with limited contrast.
Thanks in advance,
Sandra

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Email: robernal-at-u.northwestern.edu
Name: Rodrigo Bernal

Organization: Northwestern University

Title-Subject: [Filtered] Vacuum Leak in JEOL 2100 TEM Holder

Message: I am having a vacuum leak with a TEM specimen holder but cannot
pinpoint it. The holder behaves OK in a vacuum pumping station, reaching
~7 x 10^-6 Torr in about 30 minutes. After this, I transfer to the TEM,
where it pumps down at the first stage of pumping in about 5 minutes.
However, after this, when I do the rotation of the holder, just before
the full insertion, the vacuum in the column becomes way too high. In
particular, the vacuum ion gage, located in the lower left panel of the
microscope, which reads the vacuum at the pole piece, goes out of scale.
The vacuum readout for the first stage of pumping remains in a good
range though. Where the vacuum leak may be? Is there a way to pinpoint
which of the two o-rings is suspect (I've exchanged them and greased
them but the leak is still there).

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Email: joseph.uknalis-at-ars.usda.gov
Name: Joe Uknalis

Organization: USDA-ERRC

Title-Subject: [Filtered] agarose pellet stain

Message: I'm suspending small pectin beads in agarose for ease of
handling through the dehydration/embedding steps.

When I embed in LR white the 'pellet' becomes transparent.

Is there anything I could add to the agarose to give it optical density
without adding artifacts in the thin section?
I presume the ethanol dehydration steps will dissolve out most dyes i
could add...

thanks

Joe

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From drugs-express3-at-vdc.vn Sun Jul 27 09:01:57 2014
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Email: christinaheard83-at-gmail.com
Name: C. Heard

Organization: Pathlogic

Title-Subject: [Filtered] Grainy TEM images

Message: I am a new EM tech at a pathology lab specific to renal tissues. My images on the TEM have
been grainy and the resolution is suffering. I have adjusted my cutting angles, slowed the ultra
microtome speed to 1mm/s, and I post stain with UA and LC for 5 minutes each. If anyone out there
has an idea of a solution to the grainy images, it would be greatly appreciated.

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Dear Listers,

I have a researcher who would like access to an SEM that can do an automated scan of particles and automated EDS of the particles. Anyone out there close to Omaha, Nebraska? This researcher is a chemist working on aerosol particles. All replies appreciated.

Tom Bargar
UNMC
Electron Microscopy Core Facility
986395 Nebraska Medical Center
Omaha, NE 68198-6395
402-559-7347
tbargar-at-unmc.edu

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Hi Vickie,

} Best primary fixative for post-embedding
} immunogold or FluoroNanogold(TM)?

the best primary fixative is definitely high-pressure freezing,
freeze-substitution (omitting or reducing OsO4), and then any suitable
follow-up technique. Following HPF/FS, Epon is not contra-indicated to
post-embedding immunogold detection (even not low amounts of OsO4!), although
quite many people believe it is. LR-White or Gold or Lowicryls might be better
- to some degree, for a certain number and kind of specimen - , but this is not
always the case.

} What is the optimum buffer to use for a PFA/very light GTA fixative for
} immunoTEM on thin sections?
I would go for HEPES or MES or anything similar, these buffers are superior to
most others, in most cases, for most applications. Do not forget that it is not
only the buffer which matters; the other ingredients need to be optimized, in
concentration and in composition, for your tissue under investigation. PBS or
TBS or whatsoever is not always the best - in fact, it might be among the
worst.

} Is sodium cacodylate contraindicated wrt. antigen recognition?
Why still using caco if anything better is around? it is not only the disposal
of caco, which may cause problems.
kind regards,
Reinhard

--
Prof. Dr. Reinhard Rachel
University of Regensburg
Centre for EM / Anatomy
Faculty of Biology & Preclin. Med.
Universitaetsstrasse 31
D-93053 Regensburg - Germany
tel +49 941 943 2837, 1720
fax +49 941 943 2868
mail reinhard.rachel-at-biologie.uni-regensburg.de
office: VKL 3.1.29

Next microscopy conferences:
http://www.imc2014.com/
18th IMC 2014 in Prague, CZ
http://www.mc2015.de
DGE - Microscopy Conference 2015, Sept 6-11, GÃ¶ttingen

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Vickie,
For chemically stabilizing material, I prefer 2% Pf 0.2% Ga in PHEM buffer.
This can be followed with light tannic acid then UA before embedding.
This should be done cold and kept cold throughout either LR White or HM20
infiltration.
If possible HPF live or lightly fixed material then AFS with a Ga UA or a Ga
UA OsO4 combo (0.2 0.2 0.1%). Then infiltrate at -50C with HM20 (a freeze
substitution machine helps)
Good luck,
Michael Delannoy

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Name: Vickie Kimler

Organization: Wayne State University School of Medicine

Title-Subject: [Filtered] Best primary fixative for post-embedding
immunogold or FluoroNanogold(TM)?

Message: Hello All,
What is the optimum buffer to use for a PFA/very light GTA fixative for
immunoTEM on thin sections?
Is sodium cacodylate contraindicated wrt. antigen recognition?

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I have an older BSD attached to the bottom of the lens on an older XL30CP
SEM donated to us from a crime lab where EDS was done for ballistics.
Pulling the BSD off, one half of the detector has a dried fim on the Si
diode, I tried to remove using 200proof ethanol even sonicating for 1-2
minutes without any luck.

I then put the BSD in the SEM and did EDS on the surface at 15kv and got the
following composition:
Al 30%
F 39%
O 17%
C 5.5%

The rest of the comp was Si and Copper from the holder

Any ideas on cleaning ?

I have no details on how the BSD was used other than it was attached, under
vac when we picked up the microscope

Thank you

Fred Hayes
AMCaT Manager
UC Davis
Chem Eng Mat Sci

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Fred,
If you think there is an adhesive involved, try hexane (gasoline) to soften
it.

Not at all sure why someone would put a film directly on the diode.

Ken Converse
owner

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

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I have an older BSD attached to the bottom of the lens on an older XL30CP
SEM donated to us from a crime lab where EDS was done for ballistics.
Pulling the BSD off, one half of the detector has a dried fim on the Si
diode, I tried to remove using 200proof ethanol even sonicating for 1-2
minutes without any luck.

I then put the BSD in the SEM and did EDS on the surface at 15kv and got the
following composition:
Al 30%
F 39%
O 17%
C 5.5%

The rest of the comp was Si and Copper from the holder

Any ideas on cleaning ?

I have no details on how the BSD was used other than it was attached, under
vac when we picked up the microscope

Thank you

Fred Hayes
AMCaT Manager
UC Davis
Chem Eng Mat Sci

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Title-Subject: [Filtered] M & M meeting 2014 Cryo FIG

Message: This is to let everyone know I have a few more spots available for lunch. The meeting will
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officers. Lunch provided. Please let me know if you will be attending! It is important to see how
much interest there is in continuing this Focused interest group.

Thanks,
Caroline Miller, Interim Leader Elect

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Proper sample preparation is perhaps the most important element in
successful AFM imaging. Techniques for sample preparation in life
sciences, material sciences, and other applications will be covered.
This livestreaming seminar will be led by Peter Eaton Ph.D (Univ.Porto -
Requimte) and Paul West Ph.D (AFMWorkshop, Inc). Attendees are welcome
to ask questions and participate live via 'chat'.

To Register: http://www.ustream.tv/channel/Atomic-Force-Microscopes

Thank you,
Pamela K. Stone
AFMWorkshop, Inc.
1434 E. 33rd Street
Signal Hill, CA 90755
www.afmworkshop.com

==============================Original Headers==============================
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From meds-discounted5-at-rr.com Wed Jul 30 12:01:51 2014
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Would like to hear comments about the Leica ACE600 sputter coater from any
users. Interested in quality of coating for HR-FESEM using conductive metal
coatings, time it takes to reach proper vacuum, what vacuum do you achieve
before starting the coating process, other pertinent comments. Offline
comments are fine.

Thanks for your time.

Stephen McCartney
Senior Research Associate
ICTAS/NCFL
1991 Kraft Dr.
Va Tech
Blacksburg, VA 24061
USA

540-231-9765-office
540-553-6954-cell

http://www.ncfl.ictas.vt.edu/

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Dear all,

Any hints as to how to handle relocation into a new building, in a
smallish area between the four main lift (elevator) shafts, and next to
the heaviest traffic corridor from the main loading dock. Building will
house 600+ scientists, etc. which gives an idea of the traffic. The
architects, and presumably our corporate overlords, appear to have locked
into the building design, and say the microscopes just have to go in this
place.

My preferred option is to insist on external specialist consultants to
design something that will elminate or at least minimise any vibration and
electromagnetic interference. Any other suggestions?

thanks,
Rosemary

Dr Rosemary White
CSIRO Plant Industry
GPO Box 1600
Canberra, ACT 2601
Australia

T 61 2 6246 5475
F 61 2 6246 5334
E rosemary.white-at-csiro.au

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Name: Anza

Title-Subject: [Filtered] Cupromeronic Blue

Message: We would like to stain proteoglycans in tissue with
cupromeronic blue for EM studies. Does anyone know which companies are
still selling cupromeronic blue?

There are many other stains used for staining proteoglycans such as
Alican Blue, Toluidine Blue, Ruthenium Red, safranin O etc? Which one
works best? Any recommendation would be appreciated.

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Rosemary,

I'm dealing with a forthcoming move myself, although not like this.
Assuming you have to move, and can't stay where you are, then first,
have the EM companies whose 'scope you have come in and do site surveys.
When the site fails for both vibration and EMF, then insist on getting
the antivibration and EMF shielding before the move.

Might even be cheaper to build a ground-level addition to the building
with a proper, isolated foundation. Since this is an "addition" and not
a change to the building design, that might be approved. Especially
after the survey failures.

Phil

} Dear all,
}
} Any hints as to how to handle relocation into a new building, in a
} smallish area between the four main lift (elevator) shafts, and next to
} the heaviest traffic corridor from the main loading dock. Building will
} house 600+ scientists, etc. which gives an idea of the traffic. The
} architects, and presumably our corporate overlords, appear to have locked
} into the building design, and say the microscopes just have to go in this
} place.
}
} My preferred option is to insist on external specialist consultants to
} design something that will elminate or at least minimise any vibration and
} electromagnetic interference. Any other suggestions?
}
} thanks,
} Rosemary
}
} Dr Rosemary White
} CSIRO Plant Industry
} GPO Box 1600
} Canberra, ACT 2601
} Australia
}
} T 61 2 6246 5475
} F 61 2 6246 5334
} E rosemary.white-at-csiro.au
--
Philip Oshel
Microscopy Facility Supervisor
Biology Department
024C Brooks Hall
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576

==============================Original Headers==============================
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Dear Anza,
most former dealing companies (like SIGMA-Aldrich or also [http://www.labome.com/product/US-Biological/C8210.html] =} "Whoops! Sorry about that. This product no longer appears to be in our catalog.") ) stopped cancelled this product in their catalogues,

[Labome: http://www.labome.com/bin/ea.pl?link=pd1619103
US Biological
P.O. Box 261
Swampscott, Massachusetts 01907
service-at-usbio.net
www.usbio.net
800-520-3011
headquarters: USA

Order No.: C8210 product name: Cupromeronic Blue xAA
Size: 100 mg List Price: 125 USD ]

but I found CUPROLINIC BLUE (and know this dye has been used also for PG-staining in T-EM), which perhaps may serve the matter too:
http://www.buyersguidechem.com/Asimil.php?suchname=cupromeronic%20blue
http://www.buyersguidechem.com/AliefAus.php?pnumm=231952483398 :

Suppliers for Cuprolinic blue: 1 listed:

{ {Chemos GmbH Germany } }
go to: http://www.buyersguidechem.com/AfirmAus.php?fnumm=04279127&pnumm=231952483398
Address Chemos GmbH
Werner-von-Siemens-Str. 3
D-93128 Regenstauf, Germany
Phone +49-9402-9336 0
Fax +49-9402-933 613
send an inquiry
Homepage www.chemos-group.com

Another possibility perhaps:
http://www.alfa-chemistry.com/cas_81207-55-8.htm
Phone: 1-201-478-8534
Fax: 1-516-927-0118
Email: info-at-alfa-chemistry.com
For product inquiries, please use our online system or send an email toinquiry-at-alfa-chemistry.com
For order information, please contactsales-at-alfa-chemistry.com
For other customer services, please contact customer-at-alfa-chemistry.com

Further information perhaps via GOOGELING:
{Cupromeronic blue} or {Cuprolinic blue}

Best regards and wishes,
Wolfgang

Wolfgang MUSS
SALZBURG, AUSTRIA

Von: zaluzec-at-microscopy.com [mailto:zaluzec-at-microscopy.com]
Gesendet: Freitag, 01. August 2014 13:56
An: Muß Wolfgang
Betreff: [Microscopy] Cupromeronic Blue [staining in EM for PG's, any source?]

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We would like to stain proteoglycans in tissue with cupromeronic blue for EM studies. Does anyone know which companies are
still selling cupromeronic blue?

There are many other stains used for staining proteoglycans such as Alican Blue, Toluidine Blue, Ruthenium Red, safranin O etc?
Which one works best? Any recommendation would be appreciated.

Thank you

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Rosemary;
Putting microscopes near an elevator would be a disaster. The room may pass EMI specs, but the large ferromagnetic mass alters the earth's magnetic field for quite a distance, so when the cars go up and down, the alignment and focus of the microscope will change and the image may move.

John Mardinly, ASU

-----Original Message-----
X-from: Rosemary.White-at-csiro.au [mailto:Rosemary.White-at-csiro.au]
Sent: Thursday, July 31, 2014 11:44 PM
To: John Mardinly

Dear all,

Any hints as to how to handle relocation into a new building, in a smallish area between the four main lift (elevator) shafts, and next to the heaviest traffic corridor from the main loading dock. Building will house 600+ scientists, etc. which gives an idea of the traffic. The architects, and presumably our corporate overlords, appear to have locked into the building design, and say the microscopes just have to go in this place.

My preferred option is to insist on external specialist consultants to design something that will elminate or at least minimise any vibration and electromagnetic interference. Any other suggestions?

thanks,
Rosemary

Dr Rosemary White
CSIRO Plant Industry
GPO Box 1600
Canberra, ACT 2601
Australia

T 61 2 6246 5475
F 61 2 6246 5334
E rosemary.white-at-csiro.au

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Email: nickmorrill-at-byu.edu
Name: Nick Morrill

Organization: Brigham Young University/Precision Membranes

Title-Subject: [Filtered] Sample preparation using track-etched membranes

Message: Hello,

I recently discovered that track-etched membranes are used in some cases
to prepare samples before viewing in the SEM. We've developed a similar
technology in our materials science research group at BYU and I'd like
to better understand what types of applications currently use the
track-etched membranes and how well they work for this purpose. We are
trying to determine what commercial applications might exist for our
technology and wondering if some of the differences between our
membranes and the track-etched membranes are of any interest. If you
have experience using them, could you please let me know the following:

Related to sample preparation for SEM imaging:

- For the preparation of which samples/sample types do you currently
use track-etched membranes? What pore sizes/diameters are used?

- How well do the track-etched membranes work for what you are
trying to accomplish? If there are any pain-points, what are they? What
would make them better?

Thanks.

Nick Morrill
nickmorrill-at-byu.edu

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Hi Vickie,
just a comment on this: a few thoughts come into my mind. Isolation of
organelles (which one?) from any kind of tissue (which one?) is always tricky
and likely to be pretty harmful. They are taken out of their native environment
(usually, they are protected inside a cell, inside a tissue), and during
isolation, you disrupt the tisssue and the cells, inevitably - first damage -
and then you centrifuge the organelles, repeatedly - second damage (most of the
microorganisms we have looked at, suffer from centrifugation; some heavily,
some a lot, some less). Most of the organelles (mito, chloropl.) are also
damaged, and you fix the isolated, i.e. damaged organelles, don't you? second,
4%FA+0.05%GA is not the most rigorous fixative; I understand, for immuno, you
want to use 'mild' fixation conditions, but accordingly, the organelles are not
rigorously fixed; best would be cryofixation + cryoTEM, or 2% GA (not good for
immuno) plus TEM at RT, although this involves air-drying!. -- Next, I
understand that you use PBS, because this is the buffer in which your
antibodies are in best condition (they are made in blood). But, your organelles
do not like PBS: this is by no means a physiological buffer for a condition
"inside the cytoplasm of the cell". Neither the pH, nor the ionic strength, nor
the ionic composition does fit. To improve this, you would have to go into a
deeper analysis (Literature?) of the composition of the cells of your tissue;
tricky!! -- Finally, 5% Uac is neither necessary (far too high, IMO; 1 or 2% is
sufficient), and by the way, why staining at all? the pH of UAc is not
favorable (4.5) for organelles, and you will see the organelles without any
staining, and the contrast between the gold and the organelles might even be
better without UAc staining. Skip this. - and at the end, after application to
the (plastic or carbon-coated) grid, you air-dry the organelles, after several
steps which are not 'optimal' - again, this will result in 'less-than-optimal'
structure preservation.
A few steps can be improved - but do not expect too much.
good luck, and kind regards!
Reinhard

--
Prof. Dr. Reinhard Rachel
University of Regensburg
Centre for EM / Anatomy
Faculty of Biology & Preclin. Med.
Universitaetsstrasse 31
D-93053 Regensburg - Germany
tel +49 941 943 2837, 1720
fax +49 941 943 2868
mail reinhard.rachel-at-biologie.uni-regensburg.de
office: VKL 3.1.29

Next microscopy conferences:
http://www.imc2014.com/
18th IMC 2014 in Prague, CZ
http://www.mc2015.de
DGE - Microscopy Conference 2015, Sept 6-11, GÃ¶ttingen

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From vemeoro-at-spdop.ru Sat Aug 2 13:40:28 2014
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Thanks for all the useful feedback!!

I could also mention that the substation serving the building, plus the
main switchroom and distributor room are all within 10 metres of the
microscopes in the current building plan! The main electrical (and
everything else) risers for this 4 (or 5, not sure, haven't seen final
plans) storey building are 2 and 4 metres away. The elevators are closer
at 2 and 3 metres away.

Fun and games!

thanks,
Rosemary

Dr Rosemary White
CSIRO Black Mountain
GPO Box 1600
Canberra, ACT 2601
Australia

T 61 2 6246 5475
F 61 2 6246 5334
E rosemary.white-at-csiro.au

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Email: vakimler-at-med.wayne.edu
Name: Vickie Kimler

Organization: Wayne State University School of Medicine

Title-Subject: [Filtered] Immunogold and Grids

Message: Hello Experts,
What is the BEST Formvar-carbon grid to use for immunogold and/or
Fluoronanogold? It appears that nickel, nickel-asbestos and gold seem to
be the winners. It is necessary to glow discharge the grids?
Please write at your earliest convenience.
I am labeling thin-sections (Lowicryl and LR White hard/medium) as well
as isolated organelles.
Best,
Vickie

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Dear Vickie,

In principle Nickel, Gold, Rhodium and even Copper grids (with caution) are all suitable, as long as the films on the grids are well made without holes and the film and grid are not damaged or bent during the incubation and transfer. For that reason: consider using a loop wide enough to hold a grid instead of using sharp-edged tweezers. Transfer of incubation solutions can be minimised by blotting.

Some considerations:
Ni-grids are more solid than the other ones, which can be helpful when going through many steps in an incubation setup. Nickel has a disadvantage, in that it influences the electron beam and may require repeated adjustment of the astigmatism. They can sometimes magnetically stick to tweezers, which can be annoying.
Gold grids are fine, especially for labelling that does not use silver enhancement, but they are soft and bend easily. If silver enhancement is used and if there is a hole in the film, you may get enhancement of your grid and debris over the sections.
Many use regular copper grids. Even though copper may become oxidised over time and by chemicals in solutions, if your films are decent, those grids can be fine as well.

Good luck!

Jan

Jan Leunissen
i: www.aurion.nl

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From kydihe-at-mtu-net.ru Tue Aug 5 02:46:09 2014
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Email: joseph.heintz-at-covance.com
Name: Joe Heintz

Organization: Covance

Title-Subject: [Filtered] Leica UC-T Issue?

Message:

Hello All,
Currently I am using a Leica UC-T and lately I have been trying to obtain thin sections from an
epoxy block, but on the return stroke the sample face has been picking up water from the trough.
After a section is made, typically with a bit of water on it, the section is lifted from the trough
during the return stroke and while a new section is made the previous section is on top of the new
section.
I've tried a few things to remedy the issue; I have changed the knife angle a degree each way,
cleaned the knife edge and backside, dried both the block and the back of the knife and then lowered
the water level but to no avail.
I also work with a Leica UC-6 (using the same diamond knife, in the same room, on the same day)
and not had the difficulties I am having now with the UC-T. An observation, while working with the
UC-6, and UC-7 at another facility, during the return stroke the block face can easily be seen since
the arm retracts, what appears to be, a millimeter or two but on the UC-T the clearance of the
specimen block and the knife blades appears to be nonexistent. Could this clearance be the cause of
the block face to grab the water from the trough? Any way to increase the clearance length? Any
suggestions would be appreciated. If any one has a UC-T manual already in PDF form and are willing
to send it my way it would also be appreciated. Thanks for your time.

-Joe

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Besides carbon and oxygen what elements would you expect to find in a fingerprint, dander, or hair? When doing failure analysis I would like to know if there are elements that I can look for and combine with other information that would point to poor handling or cleanliness vs some other cause.

Jonathan Abbott

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From eoeuoae-at-merope.org Fri Aug 8 16:03:01 2014
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Besides C and O, Cl, Na, and K are common. N and P are not uncommon but generally at low levels.

Warren Straszheim

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Besides carbon and oxygen what elements would you expect to find in a fingerprint, dander, or hair? When doing failure analysis I would like to know if there are elements that I can look for and combine with other information that would point to poor handling or cleanliness vs some other cause.

Jonathan Abbott

==============================Original Headers==============================
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Other hobbyists and I are trying to gather up schematics, users manuals,
etc for our SEMs (ex: my Super IIIA). I'm particularly interested in
ISI SEMs. The ISI schematics we've gathered so far can be found here:
http://siliconpr0n.org/wiki/doku.php?id=microscope:sem:isi:schem

A general material list can be found here:
http://siliconpr0n.org/wiki/doku.php?id=microscope:sem:start

Please let me know if you have any material you can contribute to the
repository.

John

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From ziyogei-at-virginm.net Sun Aug 10 04:22:24 2014
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Listers,

So, this isn't a microscopy question per se, but I always get what I need from you all, so I'm going to ask it here.

Does anyone know where (or even if) it is possible to buy substrates -- single and polycrystal -- of complex oxides? I'm thinking in terms of pyrochlores like Ti2Y2O7 and other middle-of-the-periodic table crystals Ti-Zr-ox, Zr-Hf-ox, etc. Carbides and nitrides (Cr23C6, TiC, TaC, TiN, TaN, etc.) would also be interesting.

Vendor responses welcome.

Chad

---------------------
Chad M. Parish, Ph.D.
Research and Development Staff Member
Materials Science and Technology Division
Oak Ridge National Laboratory
Phone: 1 865 574 0092
Email: parishcm-at-ornl.gov

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Email: tsholst-at-5x25.com
Name: Tobias Holst

Title-Subject: [Filtered] JEOL 1200EX asterisk mode

Message: On JEOL 1200EX, after entering 'asterisk mode' with CTRL+A, there are some commands that I
am interested to know the function of:

- typing P or Q followed by a hexadecimal digit (0 to 9, A to F) displays a three byte value on the
screen which differs for each input. This is also the case when one of 00, 60 or 80 is typed in. It
appears to be modifiable by overtyping.

- typing one of CL, DA, EX, SC, ST or VA followed by ENTER causes an error but also emits a beep
rather than the usual silence. I suspect that this indicates an incomplete command, as I know VAC to
be the command for setting the vacuum system type. But I am curious to know if the others have any
function.

Does anyone know what the purpose of these may be? They do not exist in the operating manual.

Sincerely,
Tobias

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From ruoyze-at-mtu-net.ru Mon Aug 11 10:31:53 2014
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Email: acruz04-at-cibnor.mx
Name: Ariel Cruz

Organization: CIBNOR

Title-Subject: [Filtered] Capacitation SEM

"7° CURSO BÁSICO DE MICROSCOPÍA ELECTRÓNICA DE BARRIDO y avances en Microanálisis por sonda de rayos
X" La Paz, Baja California Sur, México

http://intranet.cibnor.mx/eplant1.php?pagID=anuncios/microscopia2014/index

Message: Hello

We are invite you to take part in the basic course of electron microscopy to be conducted in the
period from 25 to 29 August 2014, for more information check the page of
CIBNOR,http://www.cibnor.mx/, in the events area, thank you and I thank you for your attention,
greetings.

Ariel Cross Head of the laboratory of electron microscopy CIBNOR

Conducted in Spanish

Paulina Meza Navarro
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Email: Tracy.Lawrence-at-inspection.gc.ca
Name: Tracy Lawrence

Organization: Canadian Food Inspection Agency

Title-Subject: [Filtered] Sticky specimen control knob

Message: The specimen control knob that moves the specimen in the y-plane has gradually become
unresponsive and now will not move the specimen to the right at all. I have a Hitachi H-7100 TEM.
Can anyone give me some ideas how to fix it?
Thank you
Tracy

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Email: djm
Name: Dean J. Miller

Organization: ANL EMCenter - NanoScience and Technology Division

Title-Subject: [Filtered] Position Open - Facility Manager ANL EMCenter

Message: The Electron Microscopy Center at Argonne National Laboratory is seeking candidates for the
position of Facility Manager. The Facility Manager will be responsible for:

- Managing day-to-day operations of the Electron Microscopy Center facilities for materials science
research.

- Maintaining and ensuring effective performance of the capabilities operated within the EMC
including TEM, SEM, FIB-SEM, ancillary equipment, and computers and software.

- Maintaining operational records for laboratory procedures, safety, user program and other
requirements.

- Assisting in training users and supporting research as an analyst.

Considerable knowledge and experience with maintenance and operation of electron microscopes and
related equipment, and materials science sample preparation for electron microscopy is expected.
Familiarity with the operational and maintenance requirements of an experimental/scientific facility
and specialized systems relative to materials science electron microscopy is desired.

Interested candidates should visit the following URL for further information and to submit an
application for the position:

http://www.anl.gov/careers/apply-job/external-applicants

Position ID # 322487

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Dear Fellow 'Listers',

On behalf of SCUR
( http://www.scur.org/content/e1163/e1164/index_ger.html )
I take the opportunity to inform you in time:

SAVE the DATE:

1st Announcement:

"42nd Annual Meeting of the Society for Cutaneous Ultrastructure Research,
SCUR, MANNHEIM/Germany, 12th-14th of March 2015"
( http://www.scur.org/content/e1319//SCUR_Flyer_MANNHEIM_12_14th_March_2015.pdf )

General Topic:
"Looking into and beyond the skin - wounds, stem cells and differentiation"

Location: Universitätsmedizin Mannheim
Conference Centre "Alte Brauerei"
Medical Faculty Mannheim, Heidelberg University
Local Organizers: S.W. Schneider, I. Hausser-Siller, D. Breitkreutz, V. Huck

You will find an update, including Call for Abstracts, Registration
and Accomodation forms, very soon in that place.

The SCUR (www.scur.org) looks forward to your participation and
welcomes your questions / feedback / requests regarding the Meeting

sending an e-mail either to the Local Host Organisation,
att. of Dr. Volker HUCK,
E-Mail: volker.huck-at-medma.uni-heidelberg.de

or:

The Secretary of the SCUR
Dr. Wolfgang MUSS (PhD)
Member of MSA
SALZBURG, AUSTRIA
e-mail: w.muss-at-salk.at

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Hi Tracy

It is a long time since I operated a Hitachi 7000 series but your problem
could be traced to what is a generic TEM problem?

The "O" ring on the specimen rod becomes dry over time, and, as this is the
bearing on which the stage movement runs, the stage in that direction will
become erratic. The control being used here is the right hand stage drive.
If this is your problem, running a finger lightly coated in a good quality
vacuum grease round the "O" ring will ease its movement. Once free the
specimen rod will move into the microscope more quickly on specimen
exchange, so take extra care!

If the problem is on the left hand stage drive take a close look at where
the vertical shaft enters the pivot joint; does this move easily?

I hope this helps but if you need more please come back to me direct.

Regards

Steve

Steve Chapman FRMS
Protrain for Consultancy and Courses in Electron Microscopy
Mobile +44 (0) 7711 606967 Fax +44 (0)1280 814007
www.emcourses.com

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Email: Tracy.Lawrence-at-inspection.gc.ca
Name: Tracy Lawrence

Organization: Canadian Food Inspection Agency

Title-Subject: [Filtered] Sticky specimen control knob

Message: The specimen control knob that moves the specimen in the y-plane
has gradually become unresponsive and now will not move the specimen to the
right at all. I have a Hitachi H-7100 TEM.
Can anyone give me some ideas how to fix it?
Thank you
Tracy

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From medications_popular13-at-intselsteel.com Tue Aug 12 10:39:43 2014
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SUBJECT: Artifacts in AFM Images, Livestream Seminar

The livestreaming AFM seminar series continues tomorrow with a
discussion on Artifacts in AFM Images.

An inability to detect artifacts vs. actual sample topography can
undermine the validity of your research and/or measurements.

Join in this brief, free seminar to review the most common artifact
issues and their sources. All participants can ask questions live.

To Register: http://www.ustream.tv/channel/Atomic-Force-Microscopes

Thank you,
Pamela K. Stone
AFMWorkshop, Inc.
1434 E. 33rd Street
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From qsyhuey-at-txun.net Tue Aug 12 17:43:41 2014
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On Aug 9, 2014, at 12:32 AM, jabbott-at-moxtek.com wrote:

} Besides carbon and oxygen what elements would you expect to find in
} a fingerprint, dander, or hair? When doing failure analysis I would
} like to know if there are elements that I can look for and combine
} with other information that would point to poor handling or
} cleanliness vs some other cause.

Dear Jonathan,
My high school chem teacher had an acronym for that: C HOPKINS
CaFe. I'm not sure what the best of these--highest concentration in
finger print and lowest in the environment--would be. I'd guess P or S.
Yours,
Bill

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

Hi Jonathan,

Hair and (probably nails) will be unusually rich in sulfur compared to other tissues including skin. Hair proteins, keratins and keratin-associated proteins (or KAPs) contain a disproportionately high level of cysteine amino acid which has sulfur.

As I understand it (getting into less certain territory, but something you could follow up) hair also absorbs elevated levels of copper and possibly other heavy metals. Copper is the main one.

Other than that it would just be the same crew as for all biologically based material that Bill mentioned below.

Also, some of these samples, especially skin and finger prints, probably have high levels of lipids that (given enough finger prints) will be something you don't want inside a high vacuum system I would imagine.

Kind regards
Duane

-----Original Message-----
X-from: wtivol-at-sbcglobal.net [mailto:wtivol-at-sbcglobal.net]
Sent: Wednesday, 13 August 2014 2:31 p.m.
To: Harland, Duane

On Aug 9, 2014, at 12:32 AM, jabbott-at-moxtek.com wrote:

} Besides carbon and oxygen what elements would you expect to find in a
} fingerprint, dander, or hair? When doing failure analysis I would like
} to know if there are elements that I can look for and combine with
} other information that would point to poor handling or cleanliness vs
} some other cause.

Dear Jonathan,
My high school chem teacher had an acronym for that: C HOPKINS CaFe. I'm not sure what the best of these--highest concentration in finger print and lowest in the environment--would be. I'd guess P or S.
Yours,
Bill

==============================Original Headers==============================
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20, 38 -- Subject: RE: [Microscopy] Re: EDS elements associated with people
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Email: allan.mitchell-at-otago.ac.nz
Name: Allan Mitchell

Organization: Microscopy Otago, Dunedin, New Zealand

Title-Subject: [Filtered] Electron microscopes on television and film

Message: Hi All

I am giving a talk to a general audience in two weeks and, as part of a light-hearted general
introduction to electron microscopy, I was hoping to include a little about the appearance of
electron microscopes on television and film.

There was some discussion on the listserver about this in early 2012 and I recall that it grew into
quite a long list of movies and TV series.

I thought I had saved the list onto my computer but cannot now find it. I was wondering if anyone
out there recalls saving the list and if, could send it to me.

What would be great also if anyone out there has any images (screen shots or similar) of scenes from
movies or TV series with people at the electron microscope. If so, would it be possible to send me
a copy of the image, or a link to it.

Many thanks

Have a great day.

Allan

PS, the talk is titled Continuity through Change, from an EM perspective. Not sure how I got myself
into that one.

Allan Mitchell
Microscopy Otago - Electron Microscopy
c/- Department of Anatomy
Otago School of Medical Sciences
P.O. Box 913
Dunedin
New Zealand

Phone (03) 479 5642 or 479 7301
Fax (03) 479 5086 or 479 7254
EM Centre http://ocem.otago.ac.nz/

Microscopy Otago is hosting the 27th New Zealand Conference on Microscopy, 2 - 4 February 2015.

Please visit http://microscopy2015.otago.ac.nz/ for more details.

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Alan

Go to the Microscopy Listserver archives and do a simple search.

http://www.microscopy.com

Left hand Menu select SEARCH engine

Select YEAR = All of 2012

Select MESSAGE BODY enter "Movie"

Submit

Cheers,

Nestor

} Email: allan.mitchell-at-otago.ac.nz
}
} Name: Allan Mitchell
}
} Organization: Microscopy Otago, Dunedin, New Zealand
}
} Title-Subject: [Filtered] Electron microscopes on television and film
}
} Message: Hi All
}
} I am giving a talk to a general audience in two weeks and, as part of a light-hearted general introduction to electron microscopy, I was hoping to include a little about the appearance of electron microscopes on television and film.
}
} There was some discussion on the listserver about this in early 2012 and I recall that it grew into quite a long list of movies and TV series.
}
} I thought I had saved the list onto my computer but cannot now find it. I was wondering if anyone out there recalls saving the list and if, could send it to me.
}
} What would be great also if anyone out there has any images (screen shots or similar) of scenes from movies or TV series with people at the electron microscope. If so, would it be possible to send me a copy of the image, or a link to it.
}
} Many thanks
}
} Have a great day.
}
} Allan
}
} PS, the talk is titled Continuity through Change, from an EM perspective. Not sure how I got myself into that one.
}
}
}
}
} Allan Mitchell
} Microscopy Otago - Electron Microscopy
} c/- Department of Anatomy
} Otago School of Medical Sciences
} P.O. Box 913
} Dunedin
} New Zealand
}
} Phone (03) 479 5642 or 479 7301
} Fax (03) 479 5086 or 479 7254
} EM Centre
} http://ocem.otago.ac.nz/

===========================================
Dr. Nestor J. Zaluzec
Argonne National Laboratory
Electron Microscopy Center
9700 S. Cass Ave
Bldg 212 / A-143
Argonne, Illinois 60439 USA
Email: Zaluzec-at-aaem.amc.anl.gov

Tel: 530-NES-TORZ (530-637-8679) has Voice Mail
Lab: 630-252-7901
Fax: 630-252-4798

iChat: Zaluzec-at-AIM.com
Skype: Zaluzec
Polycom: 146.139.72.119
TPM: http://tpm.amc.anl.gov

Senior Scientist - Argonne National Laboratory
Fellow of the Microscopy Society of America
Senior Fellow the Computational Institute - University of Chicago
E.P. Wigner Fellow - Oak Ridge National Laboratory
Past President Microscopy Society of America
Adjunct Professor of Physics - Northern Illinois University &
the University of Illinois at Chicago
Visiting Professor of Microscopy - Manchester University

===========================================
TPMLab: http://tpm.amc.anl.gov
MMSite: http://www.amc.anl.gov
===========================================

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

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

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Dear Allan,

Titles of MSA-Listserver-Messages (1,2,3 = number of all messages -including original question- in my files)
} TEM in "Avatar" ? { (5) (started Wedn 27.01.2010 20:53)
} EMs in the Movies { (20) (started Mo,23.01.2012 23:01)
} Updated Movie List { (1) (= [Microscopy] EMs in the Movies ) (Di 24.01.2012 00:01), which I copied and pasted here for your convenience:

======================================================
"So far thanks to over 60 responses (and a quick trawl of the archives), this is great keep them coming!
We are up to the following:

1. Flash Gordon
2. Independence Day
3. The Man in White Suit
4. Predator 2
5. CSI
6. Spiderman 2
7. Blade Runner
8. NCIS
9. Dexter
10. Contagion
11. Andromeda Strain The Andromeda Strain (1971)and then the remake 2008 tv-miniseries.
12. Quincy
13. X-Files
14. Pres Obama at Intel's Titan
15. Spiderman 1
16. American Pickers
17. The Last Mimzy
18 ALTO MEDIA 'travel' into the matter.
19. special effects for one of the Star Trek
20. Batman
21. Jurassic Park
22. The Bone Collector" analyzes something
23. Avatar
24. The Prisoner (British TV series)
25. The Naked Gun

Thanks for all the responses so far! Once collected I will put together a website and send the link to the list.
John (from john.mitchels-at-gmail.com ) I see a kind of duplicity in names here (:-))
========================================
NB: this list has been completed with other titles in the message thread: " EMs in the Movies"(see above)

So you might try to contact {John Mitchels} , perhaps he has in file (over 60 responses never were displayed or sent via the MSA-Listserver).

If you would like and give permission to send to you the original Messages from MSA-Listserver as .msg -forwarding to your email-Address, please just reply to me and advise or request. This is an action of 3 min. only.

very best regards and good luck....for the lecture/presentation, as well as because of your statement: "not sure how I got myself into that one"...

Wolfgang
(retiring most likely in December 2014 with no successor in the EM-Lab, which presumably will be shut down afterwards)

Wolfgang MUSS
Member of MSA
SALZBURG, AUSTRIA

==============================================================================================================
Von: microscopylistserver-noreply-at-microscopy.com [mailto:microscopylistserver-noreply-at-microscopy.com]
Gesendet: Mittwoch, 13. August 2014 14:26
An: Muß Wolfgang
Betreff: [Microscopy] Electron microscopes on television and film and: Microscopy Otago is hosting the 27th New Zealand Conference on Microscopy, 2 - 4 February 2015.

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Hi All

I am giving a talk to a general audience in two weeks and, as part of a light-hearted general introduction to electron microscopy, I was hoping to include a little about the appearance of electron microscopes on television and film.

There was some discussion on the listserver about this in early 2012 and I recall that it grew into quite a long list of movies and TV series.

I thought I had saved the list onto my computer but cannot now find it. I was wondering if anyone out there recalls saving the list and if, could send it to me.

What would be great also if anyone out there has any images (screen shots or similar) of scenes from movies or TV series with people at the electron microscope. If so, would it be possible to send me a copy of the image, or a link to it.

Many thanks

Have a great day.

Allan

PS, the talk is titled Continuity through Change, from an EM perspective. Not sure how I got myself into that one.

Allan Mitchell
Microscopy Otago - Electron Microscopy
c/- Department of Anatomy
Otago School of Medical Sciences
P.O. Box 913
Dunedin
New Zealand

Phone (03) 479 5642 or 479 7301
Fax (03) 479 5086 or 479 7254
EM Centre http://ocem.otago.ac.nz/
Microscopy Otago is hosting the 27th New Zealand Conference on Microscopy, 2 - 4 February 2015.
Please visit http://microscopy2015.otago.ac.nz/ for more details.

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Hi everyone,
We are using a JEOL 2100F TEM. I have 3 questions!
1. What is "Angle" correction? (it is shown on "Alignment" options and
grouped along "Tilt" and "shift", when "Angle is wrong my tilt wobbler
will wobble in oval trajectory rather than on single line)
2. How badly can it affect resolution of STEM? (in other words, how
important it is)
3. Shall I align it in Tilt or Shift mode? as when i align it in one
of modes, it gets disoriented in other mode.
Thank you
Amit

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

1) The tilt and shift alignments are making sure that you get pure tilt when you tilt and no beam shift and pure shift when you shift and no tilt. The reason that you have to do this is because they use the same sets of deflection coils to do both. For the 2100F, when correcting tilt in TEM image mode with the beam at crossover, you do X only with X-tilt wobbler and then Y only with Y-tilt wobbler. When you use the each deflector, it will move the two beams across the screen and you can bring them back to the center using the corresponding X or Y beam shift control. Of course, you stop when they are on top of each other. You do the Shift correction in diffraction mode with the condenser fully counter clockwise and the diffraction focused to a caustic spot and brought to the center of the screen with PLA control. Again, shift X wobbler with X only and shift Y wobbler with Y only. If you are doing the Tilt correction, X or Y, and the two spots are offset and go past each other, i.e. you can't get them to coincide, then you have to make the Angle correction. Adjust the two spots so that they are perpendicular to the direction of travel during the adjustment. (This should be the closest approach to each other.) Then switch to Angle with the X or Y wobbler still on and correct this offset, again, only using the same X or Y deflector knob. Once you get that set, it just doesn't change over months. The shift corrections on my instrument never seem to wonder very much at all and they line up. I don't know whether the Angle should be used in this mode or not. I've never had to do it.

2) Before aligning the STEM, you have to align the TEM. I assume that not having the TEM properly aligned would be bad for STEM. You are using beam deflection coils in STEM and you have to have the tilt and shift compensated.

3) As I said above, I've only seen it occur in Tilt correction and not the Shift correction procedure on my system. I will send you my alignment procedure in a separate email from work. If they are split when doing the shift correction, you might consider asking your JEOL service engineer about it. We've had the PM done several times since I've taken over the instrument and it is always perfect.

I hope this helps.

-Scott

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

X-from: "amit welcomes u" {amit.welcomes.u-at-gmail.com}
To: "Walck, Scott" {S.Walck-at-comcast.net}
Sent: Thursday, August 14, 2014 3:36:45 AM

Hi everyone,
We are using a JEOL 2100F TEM. I have 3 questions!
1. What is "Angle" correction? (it is shown on "Alignment" options and
grouped along "Tilt" and "shift", when "Angle is wrong my tilt wobbler
will wobble in oval trajectory rather than on single line)
2. How badly can it affect resolution of STEM? (in other words, how
important it is)
3. Shall I align it in Tilt or Shift mode? as when i align it in one
of modes, it gets disoriented in other mode.
Thank you
Amit
Â
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On Aug 13, 2014, at 5:51 AM, microscopylistserver-
noreply-at-microscopy.com wrote:

}
} Email: allan.mitchell-at-otago.ac.nz
} Name: Allan Mitchell
}
} Organization: Microscopy Otago, Dunedin, New Zealand
}
} Title-Subject: [Filtered] Electron microscopes on television and film
}
} Message: Hi All
}
} I am giving a talk to a general audience in two weeks and, as part
} of a light-hearted general
} introduction to electron microscopy, I was hoping to include a
} little about the appearance of
} electron microscopes on television and film.
}
} There was some discussion on the listserver about this in early 2012
} and I recall that it grew into
} quite a long list of movies and TV series.
}
} I thought I had saved the list onto my computer but cannot now find
} it. I was wondering if anyone
} out there recalls saving the list and if, could send it to me.
}
} What would be great also if anyone out there has any images (screen
} shots or similar) of scenes from
} movies or TV series with people at the electron microscope. If so,
} would it be possible to send me
} a copy of the image, or a link to it.
}
} Many thanks
}
} Have a great day.
}
} Allan
}
} PS, the talk is titled Continuity through Change, from an EM
} perspective. Not sure how I got myself
} into that one.
}
}
}
}
} Allan Mitchell
} Microscopy Otago - Electron Microscopy
} c/- Department of Anatomy
} Otago School of Medical Sciences
} P.O. Box 913
} Dunedin
} New Zealand
}
} Phone (03) 479 5642 or 479 7301
} Fax (03) 479 5086 or 479 7254
} EM Centre http://ocem.otago.ac.nz/
}
} Microscopy Otago is hosting the 27th New Zealand Conference on
} Microscopy, 2 - 4 February 2015.
}
} Please visit http://microscopy2015.otago.ac.nz/ for more details.
}
}
Dear Alan,
Blade Runner has not only electron microscopy, but one of David
Scharf's images, which is alleged to be a snake scale.
Yours,
Bill

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Email: lyle.gordon-at-pnnl.gov
Name: Lyle Gordon

Organization: Pacific Northwest National Lab

Title-Subject: [Filtered] Looking for an old/slightly broken TEM cryo holder

Message: Dear Microscopists,

I wanted to thank all the listserv members for their assistance with my
previous inquiries. I've just started a post-doc working on in situ TEM at
low temperatures I'm looking for an old or slightly broken LN2 cryo-cooling
holder (any manufacturer) compatible. I would prefer a holder compatible
with Philips/FEI TEMs but I might be able to work with other types. Maybe
you have something in your lab from an old CM200 gathering dust or another
holder that is broken or no longer used. I would greatly appreciate any
donations of old junk and we might be able to purchase equipment if you
have something you are looking to sell that is no longer used.

Please reply off list.

Thanks very much,
Lyle

--
Lyle Gordon
Pacific Northwest National Lab
http://lylegordon.ca

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From bfylrynve-at-psqonline.org Tue Aug 19 14:40:08 2014
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Email: swallace-at-ambientgroup.com
Name: Sean Wallace

Organization: Ambient Group, Inc

Title-Subject: [Filtered] Old Noran Detector on Hitachi 600AB -Hardware Issue

Message: Hello,
we have an issue with our detector (Model 405B-3SST) and we've recently unmounted it from the TEM.
Upon examination, the "nose" of the detector is wrapped in a material that looks like a thin foil.
There is evidence of loss of this material also, as evidenced by flakes of "foil" sitting on the
column as if the nose was perhaps making contact with internal parts and scraping off material. I
have photos I can forward to anyone interested.

First question: What is this foil material and is it necessary? It looks like some may have flaked
off and was the cause of beam blockage.
I've been told that a silver epoxy has been used on some model of TEM detectors to adhere the
Det Â“collimatorÂ” snout onto the Det tube end and this is my initial speculation

Second: would the lack of this foil material cause the detector to not operate?

I'm hoping the loss of our counts was due to simple physical blockage by the stray material. Of
course the alignment issue now comes into play as these things should not be touching anything.

any comment would be appreciated.

Sincerely,
SW

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Email: nicholas.ritchie-at-nist.gov
Name: Nicholas Ritchie

Organization: NIST, DOC

Title-Subject: [Filtered] SDD XEDS detectors going wonky

Message: Recently I've had a spat of failures of various different silicon drift detectors and I'm
wondering if my experience is typical. The detectors have been failing catastrophically with the
symptom of high fast discriminator counts leading to 100% dead time (probably low energy noise).
One day they work at spec, the next day they have failed with no obvious cause.

I'm wondering about other people's experiences? BOTH GOOD AND BAD. How long are your silicon drift
detectors lasting? How do they fail?

Please respond to me directly at nicholas.ritchie-at-nist.gov as I don't want to slander any vendors.
I'll collect the information and make it available in manner that doesn't identify vendors but will
still provide value to the community.

EDS Vendor: (like Noran/Bruker/EDAX)

Module manufacturer: (like KETEK, Amptek, PN Detectors, Seiko, or unknown)

Lifetime date:
Failed after ?? months / Still operating after ?? months.

Failure mode: (what were the symptoms? gradual/catastrophic?)

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Email: glasser-at-mpip-mainz.mpg.de
Name: G.Glaßer

Organization: Dipl.Ing.(FH) / MPI-P, Mainz

Title-Subject: [Filtered] SEM Hitachi SU80xx: flash current too low after power outage?

Message: Hi,
i am hoping to get some help/info's from C-FEG user's (... best would be Hitachi S-4800 or SU80xx
users): i have been recently on a vacation for 2,5 weeks; when returning to my workplace i was
informed that there has been a power outage three days before my return. For this reason the
instrument was completly powerless for this time period.

After resetting the power braker and doing the runup procedure according to the manual the IGP's
were back to "perfect" vaccuum within a day (#1 close to end of display range, which means
"0.0"x10^-08Pa; #3 only slightly worse.

I deceided to do the flashing manually and discovered the following: instead of "before vacation"
flash current rising to ~35µA it has been going for the last three days only to 13µA; the timespan
and the amplitude of the measured vacuum IGP1 during the flashing did not change: it goes up to
~6.5x10^-08Pa and decays within in a few seconds to less than 1x10^-08Pa

Imaging seems to be working "normal" ... but isn't this change strange?

best regards
G.Glaßer

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Email: sarah-at-biobus.org
Name: Sarah Weisberg

Organization: BioBus

Title-Subject: [Filtered] SEM prep for tardigrades (water bears)

Message: Hi,

I run a project called the BioBus, which is a mobile microscopy facility that we use primarily for
teaching at K-12 schools. We have been lent the Hitachi 3030 tabletop SEM, and are trying to prepare
tardigrades (water bears) for imaging. I have been having trouble finding a good protocol.

Has anyone ever prepared tardigrades for SEM imaging? If so, could you please send me a protocol?

Thanks,
Sarah Weisberg
BioBus Program Director

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Email: vitha-at-tamu.edu
Name: Stan Vitha

Organization: Texas A&M University

Title-Subject: [Filtered] User manual for AO-925 microtome knife sharpener?

Message: Hallo,
I just took possession of an old American Optical knife sharpener. the machine is complete,
including the plates and abrasives, but is missing the user manual. if anybody has a manual for
this or similar model, would you mind sending a PDF or paper copy my way?

Online search was only partially successful 9a half of a manual was found).

Thank you in advance!

Best regards,

Stan Vitha
Microscopy and Imaging Center
Texas A&M University

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Email: Tracy.Lawrence-at-inspection.gc.ca
Name: Tracy Lawrence

Organization: Canadian Food Inspection Agency

Title-Subject: [Filtered] Wanted: Chiller for Hitachi H-7100 TEM

Message: Hello All
I am looking for a used water-cooled chiller for our TEM.
Must be 208V 60Hz and with a cooling capacity of 23,000 BTU/hr. Please contact me outside the
listserv if you have one that might be suitable. Thank you

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From talureu-at-leemathews.com Thu Aug 21 00:38:46 2014
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} -----Ursprüngliche Nachricht-----
} Von: Muß Wolfgang
} Gesendet: Donnerstag, 21. August 2014 10:37
} An: microscopy-at-microscopy.com LISTSERVER MSA
} (microscopy-at-microscopy.com)
} Betreff: [Microscopy] Re: User manual for AO-925 microtome knife
} sharpener?
}
} Dear Stan,
} I only can subserve with an {AO 935 manual} ..this is approx. 8MB..if
} you want to receive it [I don't want to fill your e-mail account
} without your permission/prior fiat, please just mail to me.
}
} best,
} Wolfgang
}
} Von: microscopylistserver-noreply-at-microscopy.com
} [mailto:microscopylistserver-noreply-at-microscopy.com]
} Gesendet: Donnerstag, 21. August 2014 03:53
} An: Muß Wolfgang
} Betreff: [Microscopy] User manual for AO-925 microtome knife sharpener?
}
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} Email:                       vitha-at-tamu.edu
} Name:                        Stan Vitha
} Organization:                Texas A&M University
} Title-Subject:               User manual for AO-925 microtome knife
} sharpener?
} Message:
}
} Hallo,
} I just took possession of an old American Optical knife sharpener. the
} machine is complete,
} including the plates and abrasives, but is missing the user manual. if
} anybody has a manual for
} this or similar model, would you mind sending a PDF or paper copy my
} way?
}
} Online search was only partially successful 9a half of a manual was
} found).
}
} Thank you in advance!
}
} Best regards,
}
} Stan Vitha
} Microscopy and Imaging Center
} Texas A&M University
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} sharpener?
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From meds-popular15-at-perfumania.com Thu Aug 21 05:07:08 2014
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Email: pierceem-at-ornl.gov
Name: Eric

Title-Subject: [Filtered] SPM Image Analysis Software

Message: Can some one give me some perspective on the value (pros and cons) of SPIP in comparison to
other SPM software available? The others I am familiar with are GWYDDION, WSXM, and Nanoscope
(Bruker analysis package). It would also be nice to get the groups perspective on these software
packages in comparison to Image J. Please note: I have seen the information on AFMHELP.COM. IÂ’m
looking for a more in-depth perspective as we are considering purchasing SPIP.

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

Greetings,

Hooke College of Applied Sciences, located in Westmont, IL, is offering a Polarizing Light Microscopy short course September 15-19, 2014. In addition to lectures, this course emphasizes hands-on training using state-of-the-art equipment.

For further PLM training details and registration information, please follow the link below:

http://www.hookecollege.com/courses/course.asp?COURSE_ID=41

Best regards-
__________________________________________________
Chris Gorman
Admissions Specialist
Hooke College of Applied Sciences
850 Pasquinelli Drive
Westmont, IL 60559-5539 USA
630-887-7100 (tel)
630-887-7412(fax)

==============================Original Headers==============================
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Assistant Professor of Biochemistry University of Wisconsin-Madison: Imaging Molecules

Breakthroughs in the imaging of molecules at increased spatial or temporal resolution have linked classical biochemistry with studies at the cellular and organismal level. The Biochemistry Department at the University of Wisconsin-Madison (www.biochem.wisc.edu) seeks to hire an Assistant Professor (tenure track) in this area. The successful candidate will utilize state-of-art imaging methods to advance important areas in biochemistry, structural biology, cell biology or physiology. Approaches include but are not limited to: cryo-EM or correlative light-EM; electron tomography; super-resolution, single molecule or light sheet microscopy; atomic force microscopy; imaging with novel fluorescent or optical approaches. We seek individuals who employ these methods to address key issues concerning biological molecules, machines, and structures. The successful candidate will be expected to develop a vigorous, extramurally-funded research program, and to participate in teaching in the Department. Attractive recruitment packages include start-up funding, salary support, state-of-art core facilities, exceptional laboratory space, access to vibrant graduate training programs, and a collegial and collaborative environment.

Applicants should submit PDF files of curriculum vitae with summary of research accomplishments, description of future research program and teaching interests, and letters from three references to jobs-at-biochem.wisc.edu by October 15, 2014.
Please note: Unless confidentiality is requested in writing, information regarding applicants must be released upon request. Finalists cannot be guaranteed confidentiality. UW-Madison is an equal opportunity/affirmative action employer.
Department

--
Kevin W. Eliceiri
Director, Laboratory for Optical and Computational Instrumentation (LOCI)
Departments Cell and Molecular Biology and Biomedical Engineering
Affiliate Principal Investigator, Morgridge Institute for Research (MIR)
Room 271 Animal Sciences, 1675 Observatory Drive, Madison, WI 53706
Phone: 608-263-6288

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Email: jhyun-at-gatan.com
Name: John Hyun

Organization: Gatan

Title-Subject: [Filtered] EELS and EFTEM Training, October 2014

Message: EELS & EFTEM Analysis Training School. October 14-17, 2014. Gatan, Pleasanton, California,
USA.

Want to refresh or advance your EELS and EFTEM knowledge? Join us for an intensive 4-day training
school that incorporates lectures, computer laboratories, and microscope practicals to provide
participants with comprehensive, hands-on training on key EELS and EFTEM topics and technology.

Online registration is now open at www.gatan.com/resources/training/EELSSchoolOct2014.php. School
fee: $1,750.00 US Dollars. Space is limited.

Overview: Transmission electron microscopy (TEM) reveals details of natural and man-made structures
at the micrometer, nanometer, and even sub-nanometer scale. Energy-filtered TEM (EFTEM) and electron
energy-loss spectroscopy (EELS) are the ideal analytical partners to the high spatial resolution
provided by TEM in both the conventional and scanned (STEM) imaging modes.

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Email: vakimler-at-med.wayne.edu
Name: Vickie Kimler

Organization: Wayne State University School of Medicine

Title-Subject: [Filtered] Chloroform Flattening of Sections

Message: When one chloroform-flattens ultrathin sections for the TEM, is there artifactual
stretching and resultant alterations in the dimensions of structures, which are critical in
morphometric work?

Does chloroform have a greater effect on LR White/Lowicryl versus the Embed and other harder
non-immuno resins?

Finally, on a serendipitous note, would chloroform have the ability, if it does indeed stretch apart
structures on a nano level, open up antigenic determinants for the often difficult immunogold work?

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From wikove-at-stv.ru Sun Aug 24 01:20:58 2014
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I am sure overstretching is a risk but so is not stretching. To get a feel for the effects of compression on thin sections, look at the article by Studer and Gnägi (2000) Minimal compression of ultrathin sections with use of an oscillating diamond knife. Journal of Microscopy, Vol. 197(1):94-100. This article is about using a vibrating diamond knife to avoid compression but I remember it having good comparison images. Diatome's web page has an interesting discussion at http://www.diatomeknives.com/knives/pdf/ultrasonic_flyer_usa_1108.pdf. They say the compression of various resins are 10-20% for epoxy resins, 12-24% for Lowicryl K4M, 10-17% for Spurr's and8-13% for LR White. I don't use chloroform anymore. I am a big fan of the less toxic heat pens. I feel 120 V electrical-line models are much better than battery-operated ones.

Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
phillipst-at-missouri.edu

http://www.biology.missouri.edu/faculty/phillips.html
http://www.biotech.missouri.edu/mcc/

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Title-Subject: [Filtered] Chloroform Flattening of Sections

Message: When one chloroform-flattens ultrathin sections for the TEM, is there artifactual stretching and resultant alterations in the dimensions of structures, which are critical in morphometric work?

Does chloroform have a greater effect on LR White/Lowicryl versus the Embed and other harder non-immuno resins?

Finally, on a serendipitous note, would chloroform have the ability, if it does indeed stretch apart structures on a nano level, open up antigenic determinants for the often difficult immunogold work?

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Title-Subject: [Filtered] Position for confocal microscopist

Message: Dear Listers,

I have an opening for an experienced confocal microscopist, preferably with experience of at least
one of the following techniques multiphoton microscopy, OPO lasers, 2nd and 3rd harmonic reduction
imaging, live cell imaging FCS and or and FLIM. A functional knowledge of TEM is also desirable.
Below is a summary of the role.

Purpose of Role:
The post holder will be responsible for the confocal microscope and other light microscopes within
the imaging group and for the training and development of the light microscopy side of the group.
The post holder will report to the imaging group manager.
Key Responsibilities:
Â• Contribution of personal technical expertise and involvement in a least one of the key resource
provisions of the section.
Â• Data acquisition, processing and interpretation on collaborative research projects.
Â• Sample preparation including: fluorescent, histochemical and immunological staining; live cell
labelling; cell and or microbiological culture techniques and standard laboratory procedures.
Â• Training of end users and the creation and update of SOPÂ’s, COSHH and risk assessmentsÂ’.
Â• Developing and managing internal collaborations.
Â• Maintenance of equipment and liaison with service engineers and company representatives.
Â• Keeping up to date with advances in the field and actively participating in courses and conferences.
Â• The option to pursue individual research interests.

Applications can be made via New Scientist, or the civil service jobs website (position 1420701)
where you will be asked to fill in a massive form (please do not let that put you off applying).

The National Institute for Biological Standards and Control (NIBSC), a Centre of the Medicines and
Healthcare Products Regulatory Agency (MHRA), is a world leader in assuring the quality of
biological medicines through product testing, developing reference materials and applied research.
Due to the nature of its role we bring together a wide range of scientists covering R&D, production
and testing of reference materials and biological medicines and have numerous state of the art
machines and techniques on site. NIBSC is located to the north of London and is surrounded by the
beautiful Hertfordshire countryside, there are nearby transport links to London and Cambridge.
Further information on NIBSC can be found at http://www.nibsc.org/ or by contacting myself.

All the best,

Kirsty

Kirsty MacLellan-Gibson

Head of Biological Imaging

National Institute for Biological Standards and Control (NIBSC)
Blanche Lane, South Mimms, Potters Bar, Hertfordshire, EN6 3QG UK
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Dear All,
anybody out who can give me a detailed protocol how to grow / fix / postfix leucocytes / T-cells on coverslips to be imaged in the
SEM?
Best would be to have them OsO4-stained also at some point...
Drying would be through ethanol to critical point drying.

Thanks,
Stefan

--

-----------------------------------------------------
Stefan Diller - Scientific Photography
Arndtstrasse 22
D - 97072 Wuerzburg Germany
++49-931-7848700 Phone
++49-931-7848701 Fax
++49-175-7177051 Mobile

Websites:
www.nanoflight.info
www.stefan-diller.com
www.electronmicroscopy.info
www.elektronenmikroskopie.info
www.zwillingsprojekt.de
www.assisi.de
Anfahrt: http://Mail.map24.com/Stefan.Diller
-----------------------------------------------------

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Stefan,

What I used for FE-SEM of lymphocytes:
Cultured cells on glass coverslips in their normal growth medium. Make
*sure* they're a monolayer, nonconfluent is preferred.
It's best to sputter-coat the coverslips first, then sterilize with
alcohol or UV. The cells like a gold or gold/palladium substrate, and
they're on a conductive surface, which makes life easier.

Fix 1-2 hrs at room temperature in 1.25% glutaraldehyde in appropriate
buffer. The buffer weaseling is because the choice can depend on what
you want to do with the cells. Sorenson's phosphate works, so does
Na-cacodylate, HEPES, etc. I used 0.1M, 0.15M might work for yours.
Add 1% monomeric tannic acid to the fix. This helps prevent holes in the
cell membranes.

OsO4 wasn't needed, but if you want to use it, then 1% OsO4 + 1%
monomeric tannic acid for 1-2 hours at room temp. Overnight in the 'frig
would probably be OK, but I'd test it first.

Dehydrate through ethanol - I started at 30%, then
50-70-80-90-95-3x100%, 5 minutes each.
CPD: 5 soaks at 3 minutes each in liquid CO2, then do the run. 5 minute
soaks can also be used.

Coat with your favorite high-resolution coater.

Phil

} Dear All,
} anybody out who can give me a detailed protocol how to grow / fix / postfix leucocytes / T-cells on coverslips to be imaged in the
} SEM?
} Best would be to have them OsO4-stained also at some point...
} Drying would be through ethanol to critical point drying.
}
} Thanks,
} Stefan
}

--
Philip Oshel
Microscopy Facility Supervisor
Biology Department
024C Brooks Hall
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576

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Research Scientist Position in the Cryo-Electron Microscopy Facility at
the New York Structural Biology Center

The New York Structural Biology Center (NYSBC) seeks an experienced
electron microscopist to join the staff of its Cryo-Electron Microscope
Facility (http://cryoem.nysbc.org). The NYSBC is a shared center that
supports state-of-the-art research in cryo-EM, NMR, and X-ray
crystallography. Cryo-EM facilities include four transmission electron
microscopes and a dual-beam scanning electron microscope, which support
projects involving electron tomography, single particle analysis and
electron crystallography of both stained and frozen-hydrated samples.
Projects focus on 3D reconstruction of biological assemblies ranging
from the atomic structure of membrane proteins, to the subunit
organization in macromolecular complexes and the cellular anatomy of
developing organisms, to 3D reconstructions of entire cells and their
distributions within their native tissue using the dual-beam microscope.
To assist in these developments, NYSBC seeks an individual with
postdoctoral experience in biological electron microscopy and image
reconstruction. This individual will carry out experiments in support of
collaborative projects with affiliated investigators and will also have
opportunities to pursue independent research projects. The individual
should be capable of multitasking, should enjoy working with other
people, should have a good working knowledge of electron microscopes and
a strong research background. Good communication skills are essential.

To apply, please send a c.v. and the names of three references to
rice-at-nysbc.org

--
William J. Rice, Ph.D.
Senior Scientist, EM Facility Manager
New York Structural Biology Center
89 Convent Avenue, NY, NY 10027

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From waksobge-at-topgunner.com Wed Aug 27 00:48:29 2014
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Let me ask a question about "stopping power." I have some unstained, thin sections of PVC mixed with nitrile rubber which I examined at 80KV in our TEM.

I would expect the chlorine containing PVC to be more opaque or have greater contrast than the nitrile, but the c triple bond N has me a little concerned about my judgment.

Does anyone have any experience with pvc/nitril morphology they would like to share?

Thanks!
Frank
ARDL

This email and any of its attachments may contain confidential information intended only for the use of the addressee(s). If the reader of this email is not the intended recipient or the employee or agent responsible for delivering it to the intended recipient, you are hereby notified that any dissemination or copying of this email is strictly prohibited. If you have received this email in error, please notify us by return email at info-at-ardl.com, permanently delete the email, and destroy any printouts. If this email contains test data and/or draft reports, you are hereby notified that only a signed original test report will contain official results, a copy of which resides in the project folder located at ARDL, Inc. Thank you. Akron Rubber Development Laboratory, Inc.

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Email: robernal-at-u.northwestern.edu
Name: Rodrigo Bernal

Organization: Northwestern University

Title-Subject: [Filtered] Z-axis orientation in JEOL 2100

Message: Hello All,

What is the positive z-axis in the stage of the JEOL 2100? Is it pointing up towards the electron
gun, or down to the phosphor screen?

When I put a sample in my custom made TEM holder, it gets focused in the -90um z coordinate, which
is too far; I want to know if I need to thin my sample support in order to bring the sample down
(towards the phosphor screen), or add a spacer to bring it up (towards the electron gun).

Thanks.

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Dear colleagues,

we are the proud owners of a Zeiss axioimager D1 coupled to a digital camera whose make I won't cite.
This system is 10 years old and the camera is not working as well as we wish. The microscope cannot be the culprit because it is made in Germany ;-). The user needs to work several more years to pay the rest of his loan and cannot afford to be replaced. So I think it is time to change the camera (scientists are pragmatic).
We don't need the super-high resolution last-model camera, it is mainly used to take pictures of agar plates for documentation (bright field).
I wondered if we couldn't just couple a low-budget digital camera from the usual store around the corner (80-100 Euros).
Did someone already try that? As a biologist I have nearly no knowledge in mechanics and I am not very experienced in photography neither (except microphotography, which doesn't help here).
I wonder if it is possible to couple a usual digital camera (not a microscope camera) to a Zeiss microscope of this type? The camera should have a USB port of course and maybe there would be some ring on the market to couple both together in perfect hamony (a little bit like ebony and ivory).
Please be kind enough to write in standard units, inches and gallons give me headaches. Pixels and megabits are OK ;-).
Many thanks in advance.

Stephane

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Email: mobradovic-at-vinca.rs
Name: Marko Obradovic

Organization: INN "VINCA", Belgrade, Serbia

Title-Subject: [Filtered] Retrofiting TEM with a digital camera

Message: Hello,

My name is Marko and I am a PHD student from Vinca Institute, Serbia.
This is my first message here, so I apologize in advance if I do
something incorrectly.
We have in our laboratory an EM400 Philips TEM, produced in 1980, which
up until recently was used for material science characterization. It is
currently being repaired and since we have some funds available I thought
it could be useful to retrofit it with a digital camera, to make it
easier to use.
So, my question is the following: which bottom-mounted CCD camera would
be the best Price-performance option for us, since the effects that we
are looking fore are in 5 to 20 nm range? And also, are there any good
used cameras for sale because our funds are somewhat limited?

Thank You all in advance,

Marko Obradovic
Research Assistant,
INN "VINCA",
11000 Belgrade, Serbia

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Email: kun.li-at-kaust.edu.sa
Name: Kun Li

Organization: King Abdullah University of Science and Technology

Title-Subject: [Filtered] Opening for TEM service engineer

Message: King Abdullah University of Science and Technology is an international graduate-level,
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of scientific achievement in the Kingdom that will also benefit the region and the world.

Key responsibilities include:
Â• Responsible for the overall maintenance of all the transmission electron microscopes in the
Imaging and Characterization Core Lab to ensure high equipment uptime.
Â• Conduct preventive maintenance of all the seven TEMs per predefined PM schedule.
Â• Conduct trouble-shooting and corrective maintenance of all the TEMs.
Â• Work with FEI factory service support team to speed up the trouble shooting and problem solving
process.
Â• Manage all the TEM spare parts and consumables.
Â• Responsible for the management and maintenance of the TEM Sample Preparation Lab.

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Â• Good knowledge of working principle and maintenance of TEM
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From wnibedu-at-gtmc.net Thu Aug 28 08:30:36 2014
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Rodrigo-

As far as I can tell, the arrows on the Z buttons correspond to the actual directions of motion. So Z} 0 indicates that the sample is above the eucentric plane.

You can find the answer by checking the objective lens current needed to bring the sample into focus. Press STD FOCUS. Then turn the Focus knob until the sample is in focus. If you turned Focus clockwise, the OL is overfocused (current larger than the standard value), so the sample is below the eucentric height. This indicates that Z will have to be increased (sample raised) to bring the sample to the eucentric height.

Look at it this way: 1/p=1/f-1/q. We're not changing the image distance q. Start off with the sample in focus: If the focal length f is increased (OL current decreased), the object distance p has to be increased to bring the object into focus. Since the sample is above the OL, this corresponds to raising the sample (increasing Z).

You can check how the OL current changes when turning the Focus knob by looking at the Status/Lens Voltage dialog. But the convention seems to be that turning clockwise increases the OL current.

-Phil

P.S. I'd be interested to know more about your custom holder.

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Email: robernal-at-u.northwestern.edu
Name: Rodrigo Bernal

Organization: Northwestern University

Title-Subject: [Filtered] Z-axis orientation in JEOL 2100

Message: Hello All,

What is the positive z-axis in the stage of the JEOL 2100? Is it pointing up towards the electron gun, or down to the phosphor screen?

When I put a sample in my custom made TEM holder, it gets focused in the -90um z coordinate, which is too far; I want to know if I need to thin my sample support in order to bring the sample down (towards the phosphor screen), or add a spacer to bring it up (towards the electron gun).

Thanks.

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Marko,

I think you have four choices of manufacturer:

- Gatan
- AMT
- OSIS
- TVIPS

They all make good cameras, with somewhat different emphases on application. As well as the quality of the camera, you also need to consider support, both with regard to service and applications.

Can I suggest you contact:

MB Science Service
Hodrusha Hamre 485
SK-96663 Hodrusha
SLOVAKIA

Mr Manfred Baumann
mbssbaumann-at-bb.telecom.sk
Tel: + 421 45 68 44 083
Fax: + 421 45 68 44 084
Mobile: + 49 (171) 448 2339

A long time ago, I used to work with Manfred - he knows what he is talking about. They are a Gatan agent but might be able to help with used cameras and should be able to provide support.

Best regards,

Larry Stoter
(Retired - https://www.linkedin.com/pub/larry-stoter/53/439/963)

On 28 Aug 2014, at 14:37, microscopylistserver-noreply-at-microscopy.com wrote:

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} Email: mobradovic-at-vinca.rs
} Name: Marko Obradovic
}
} Organization: INN "VINCA", Belgrade, Serbia
}
} Title-Subject: [Filtered] Retrofiting TEM with a digital camera
}
} Message: Hello,
}
} My name is Marko and I am a PHD student from Vinca Institute, Serbia.
} This is my first message here, so I apologize in advance if I do
} something incorrectly.
} We have in our laboratory an EM400 Philips TEM, produced in 1980, which
} up until recently was used for material science characterization. It is
} currently being repaired and since we have some funds available I thought
} it could be useful to retrofit it with a digital camera, to make it
} easier to use.
} So, my question is the following: which bottom-mounted CCD camera would
} be the best Price-performance option for us, since the effects that we
} are looking fore are in 5 to 20 nm range? And also, are there any good
} used cameras for sale because our funds are somewhat limited?
}
} Thank You all in advance,
}
} Marko Obradovic
} Research Assistant,
} INN "VINCA",
} 11000 Belgrade, Serbia
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Email: freth001-at-umn.edu
Name: Chris Frethem

Organization: University of Minnesota Characterization Facility

Title-Subject: [Filtered] Humidity / frosting problems in cryoEM lab

Message: Our labs are contending with high humidity problems during cryoultramicrotomy, cryoTEM (&
Viotrobot sample preps), cryoSEM, etc. Midwestern summer weather is a challenge. Colder weather much
less so.
Air supply is whole-building; i.e. common source & exhaust, with some local zone control via
variable air volume valves. Facilities Management has lately been able to keep temperatures in
68-73F range; relative humidity mostly in 48%-60% range.
Facilities Management requested that I seek ideas from other facilities to improve situation. I
suspect we'd have to isolate the facility from rest of building; probably very expensive. Adding
dehumidifiers would add heat & noise (vibrations) to rooms but seem pointless if all building air is
shared. Likewise we must preserve good air quality, i.e sufficient turnover.
Understood that proper sample techniques and instruments and instrument use are key factors; just
looking for other ideas from those similarly challenged. Thank you.

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Chris,

I just read your message and thought about how my glass of ice water last
night had so much water on the outside. What if you put several beakers of
ice on the other side of the room or maybe in your sink, would enough
water be removed from the air by condensation to help?

Years ago I had problems with summer humidity causing too many holes in my
collodion films for the grids that I was attempting to coat. I found I had
to make them beside a running area heater. Hot and dry! It worked fine for
me then I needed the ice water. I do question it for cryoEM however.

Opinions and experiences related are those of Pat Connelly and do not
represent the NIH. This message is not confidential and
can be freely shared and reproduced.

Pat

Patricia Stranen Connelly
Research Assistant
NHLBI EM Core Facility
National Institutes of Health
Building 14E Room 111B
Bethesda, MD 20892-5570
301-496-3491
connellyps-at-mail.nih.gov

http://www.nhlbi.nih.gov/research/intramural/researchers/core/electron-micr
oscopy-core/index.html

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Hi Chris,

} Our labs are contending with high humidity problems during cryoultramicrotomy, cryoTEM (&
} Viotrobot sample preps), cryoSEM, etc. Midwestern summer weather is a challenge. Colder weather much
} less so.
} Air supply is whole-building; i.e. common source & exhaust, with some local zone control via
} variable air volume valves. Facilities Management has lately been able to keep temperatures in
} 68-73F range; relative humidity mostly in 48%-60% range.

I was once facing a similar problem, aggravated by the face that the health and safety inspector had demanded a humidifier (!) for the building.

The high rate of air exchange (8 times per hour, as far as I remember) in the lab precluded the use of mobile dehumidifier units and any other provisional solutions. In the end, a large, fixed dehumidifier was installed on the corridor in the incoming air supply of three rooms - cryopreparation, Polara, and another microscope. Yes, this unit IS noisy (hence on the corridor) and using a lot of energy, but it is worth it!

Please let me know if you need any further details like manufacturer.

Best regards,

Guenter

--
Dr. Guenter Resch
Nexperion e.U. - Solutions for Electron Microscopy - www.nexperion.net
Mattiellistrasse 3/17, 1040 Vienna, Austria - Phone +43 664 94 17 210
Registered at Commercial Court Vienna, FN 397677w - VAT Reg. ATU67962234

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Dear Chris,
Our solution to temperature and humidity control for an aberration corrected
STEM (though not cryo) is a dedicated, semiconductor industry type A/C unit.
They can be small enough for just a room - though they do need their own
closed cycle duct work. Temperature is very precise and maximum allowable
humidity and air flow rate are set independently. Totally quiet and stable
in the room, as they are mounted remotely. Does not need venting to the
outside. It is a more expensive solution, perhaps, but not compared to the
price of the microscope or your lost productivity. I can pass along more
details if you are interested.
Regards,
Larry Scipioni

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Email: freth001-at-umn.edu
Name: Chris Frethem

Organization: University of Minnesota Characterization Facility

Title-Subject: [Filtered] Humidity / frosting problems in cryoEM lab

Message: Our labs are contending with high humidity problems during
cryoultramicrotomy, cryoTEM (&
Viotrobot sample preps), cryoSEM, etc. Midwestern summer weather is a
challenge. Colder weather much
less so.
Air supply is whole-building; i.e. common source & exhaust, with some local
zone control via
variable air volume valves. Facilities Management has lately been able to
keep temperatures in
68-73F range; relative humidity mostly in 48%-60% range.
Facilities Management requested that I seek ideas from other facilities to
improve situation. I
suspect we'd have to isolate the facility from rest of building; probably
very expensive. Adding
dehumidifiers would add heat & noise (vibrations) to rooms but seem
pointless if all building air is
shared. Likewise we must preserve good air quality, i.e sufficient turnover.
Understood that proper sample techniques and instruments and instrument use
are key factors; just
looking for other ideas from those similarly challenged. Thank you.

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Email: brian.choi-at-parkafm.com
Name: Brian Choi

Organization: Park Systems

Title-Subject: [Filtered] Tissue sample preparation for SEM

Message: Dear whom has the know-how,

I tried to acquire the surface morphology of large intestin with SEM.
In the SEM image I obtaiend, the surface of large intestin tissue is fully covered with some stiky
muscus so that I can barely see the specific surface pattern of intestin.

Is there any treatment method to get rid of the muscus on the surface?

I will be very pleased if you inform me any protocol for this.

Thank you very much

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What systems are currently being made for ion-beam coating for
high-resolution FE-SEM.
I know of the Gatan coater, and South Bay Technologies (and the osmium
coater from SPI). What else is there?
Thanks.
--
Philip Oshel
Microscopy Facility Supervisor
Biology Department
024C Brooks Hall
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576

==============================Original Headers==============================
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Dear Listers,
in our SEM the condenser polar pieces have a couple of O-rings each
around their body. Given that above and below the whole condenser
section O-rings are present around the column bore, I wonder if those
O-rings around the bodies of condenser polar pieces may affect the
"general" vacuum level, or if they are aimed to something else, for
instance keeping cleaner a particular area of the polar piece external
surface.
Any idea?
Thanks

Davide

~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~~~~~~~
Davide Cristofori

direct phone = 041.234.67.26
e-mail = dcristofori[at]unive[dot]it

Università Ca' Foscari Venezia
Dipartimento di Sc. Molecolari e Nanosistemi
Via Torino, 155b I-30172 Mestre (VE) Italy
~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~~~~~~~

==============================Original Headers==============================
5, 20 -- From dcristofori-at-unive.it Sat Aug 30 02:29:58 2014
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From uzenebo-at-kubangsm.ru Sat Aug 30 03:47:03 2014
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Dear Davide,

Your SEM may be a JEOL. This couple of O-rings was designed for vacuum
only. When the current flow inside the lens coil this pole piece is
strongly set in position by the resultant magnetic field. This position
is fixed by the mechanical design of the top of the pole piece and the
associated screws. When contamination occurs here it comes from inside
(gun pipe for example) and from the pumping system the more often. It
may be better to use a good grease for those O-rings and a small amount
of it to avoid contamination from O-rings.

--
Nicolas STEPHANT

Université de Nantes
Institut Jean Rouxel
Service de microscopie électronique à balayage et microanalyse
2 rue de la Houssinière
BP 92208
44322 Nantes cédex 3

Tél : 02 40 37 64 26
Mél : nicolas.stephant-at-univ-nantes.fr
Web : http://cnrs-imn.fr

"Le monde n'existe que pour autant que nous sommes capables d'en produire une image"
C.G Jung

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Dear Listers,
in our SEM the condenser polar pieces have a couple of O-rings each
around their body. Given that above and below the whole condenser
section O-rings are present around the column bore, I wonder if those
O-rings around the bodies of condenser polar pieces may affect the
"general" vacuum level, or if they are aimed to something else, for
instance keeping cleaner a particular area of the polar piece external
surface.
Any idea?
Thanks

Davide

~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~~~~~~~
Davide Cristofori

direct phone = 041.234.67.26
e-mail = dcristofori[at]unive[dot]it

Università Ca' Foscari Venezia
Dipartimento di Sc. Molecolari e Nanosistemi
Via Torino, 155b I-30172 Mestre (VE) Italy
~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~~~~~~~

==============================Original Headers==============================
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Dear listers,

a big THANK YOU for those who answered my question about cheap solutions to couple a camera to a Zeiss LM.
A got several interesting Emails, one specially from James, who told me about his experience with a Touptek EM-510C camera.
I searched the web for infos about this camera and found lots of positive opinions, in particular in a german amateur forum. It seems to suit my needs and I found the camera in Ebay and Amazon for an incredible $179 (around 130 Euros). It is definitely not a high-end camera, don't misunderstand me but I am not looking for one neither.
It has a C-mount and a USB port and the software seems to be appreciated (free to download on the company website). I think it would be a mistake not to try it when you think an ELISA kit alone costs around 500 Euros.
I have no financial interest in this product, I just thought it would be an interesting information for other people.

a big NOT-THANK YOU for the registered forum users who already sent me and who will send me an automatic out-of-office reply. This is not funny. Seriously.

Have a nice and productive camera/week/life (optimally all 3).

Stephane

==============================Original Headers==============================
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From dyvvawe-at-unitymediagroup.de Mon Sep 1 03:40:59 2014
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Dear All,

has anybody experience with the substitute of Kodak D-19 developer for
Kodak microscopy film 4489? I bought a package from Plano (Ted Pella)
for 1 gallon. But it is not clear to me if it is for preparing a stock
solution or the working solution and is the handling exactly the same
as for the old Kodak D-19? I really appreciate any comment.

Best regards,
Anne
Dr. Anne Heller
AG Elektronenmikroskopie
Institut fÃ¼r Botanik (210)
UniversitÃ¤t Hohenheim
Postfach, 70593 Stuttgart
Tel. 0711-45922180
Fax. 0711-45923355

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The U.S. Department of Agriculture, Agricultural Research Service has a Full Time Permanent Biologist position GS 9/11 available for a transmission electron microscopy specialist with the Electron & Confocal Microscopy Unit in Beltsville, MD. The job announcement number is: CK-14-1181279-TW. See the web page for full details. https://www.usajobs.gov/GetJob/ViewDetails/378945700. Position is open Sept. 2 through Sept. 15, 2014.

The purpose of this position is to provide support in a core microscopy facility providing expertise in transmission electron microscopy (TEM). The position involves preparation of various organisms, tissues, and materials from plants, animals and microbes for observation in a TEM, operating a TEM to produce high quality images, and instruct and supervise scientists to use the TEM. The position is located in the Electron and Confocal Microscopy Unit within the Soybean Genomics and Improvement Laboratory at the Beltsville Agricultural Research Center whose general mission is to provide collaborative research studies using electron, confocal, and light microscopy techniques to study plants, animals and microbes through the development of high quality and high resolution images for publication of research. Go to: www.usajobs.gov

Gary R. Bauchan Ph. D
Director, Electron & Confocal Microscopy Unit
USDA-ARS
10300 Baltimore Ave.
Bldg. 012, 5th St., BARC-West
Beltsville, MD 20705
301-504-6649
gary.bauchan-at-ars.usda.gov

This electronic message contains information generated by the USDA solely for the intended recipients. Any unauthorized interception of this message or the use or disclosure of the information it contains may violate the law and subject the violator to civil or criminal penalties. If you believe you have received this message in error, please notify the sender and delete the email immediately.

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Anne,

We use the replacement D-19 here. It seems to be easier to over-develop
with the Photographer's Formulary version than with the Kodak D-19. But,
it needs to be tested.
Curiously, none of the EM supply companies I've spoken with have
bothered to do this, they all say "it's a straight replacement, use it
like you did Kodak D-19." Which is not unreasonable, given that the PF
version is the same chemicals in the same proportions as the Koday
version, but still ...

The replacement D-19 is compounded by Photographer's Formulary
http://stores.photoformulary.com/formulary-substitute-d-19/

This is the same mixture as Kodak D-19, but packaged as separate
components, which have to be mixed in a specific order when making the
stock D-19 solution - directions are in the box.

Phil

} Dear All,
}
} has anybody experience with the substitute of Kodak D-19 developer for
} Kodak microscopy film 4489? I bought a package from Plano (Ted Pella)
} for 1 gallon. But it is not clear to me if it is for preparing a stock
} solution or the working solution and is the handling exactly the same
} as for the old Kodak D-19? I really appreciate any comment.
}
} Best regards,
} Anne
} Dr. Anne Heller
} AG Elektronenmikroskopie
} Institut fÃ¼r Botanik (210)
} UniversitÃ¤t Hohenheim
} Postfach, 70593 Stuttgart
} Tel. 0711-45922180
} Fax. 0711-45923355

Philip Oshel
Microscopy Facility Supervisor
Biology Department
024C Brooks Hall
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576

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Greetings,

Hooke College of Applied Sciences, located in Westmont, IL, is offering a Transmission Electron Microscopy short course October 7-9, 2014. In addition to lectures, this course emphasizes hands-on training using state-of-the-art equipment.

For further TEM training details and registration information, please follow the link below:

http://www.hookecollege.com/courses/course.asp?COURSE_ID=53

Best regards-
__________________________________________________
Chris Gorman
Admissions Specialist
Hooke College of Applied Sciences
850 Pasquinelli Drive
Westmont, IL 60559-5539 USA
630-887-7100 (tel)
630-887-7412(fax)

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Hi

For those who are intersted (and have some time for that stuff), here is
the recepee of the D19b :

Grammes for 1l final solution. To be diluated at 30-40 °C in distilled
water to make a concentrated mother solution at 1/4 or 1/3.
This mother solution will be kept in an air and light free bottle at {
18°C and diutated as wanted for use.

Génol (Métol, Rhodol, Elon) 2.2
Sulfite de Sodium 72
Hydroquinone 8.8
Carbonate de Sodium 48
Bromure de potassium 4

Sorry, it's with french names, but it should be easy to find the
translations.

Hoping it can help someone !

Jacques

--

J. Faerber
IPCMS-DSI
Institut de Physique et Chimie des Matériaux de Strasbourg
Département Surfaces et Interfaces
23, rue de Loess ; BP43
67034 Strasbourg CEDEX 2
France

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

Le 02/09/2014 11:55, Anne.Heller-at-uni-hohenheim.de a écrit :
} ----------------------------------------------------------------------------
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} ----------------------------------------------------------------------------
}
}
} Dear All,
}
} has anybody experience with the substitute of Kodak D-19 developer for
} Kodak microscopy film 4489? I bought a package from Plano (Ted Pella)
} for 1 gallon. But it is not clear to me if it is for preparing a stock
} solution or the working solution and is the handling exactly the same
} as for the old Kodak D-19? I really appreciate any comment.
}
} Best regards,
} Anne
} Dr. Anne Heller
} AG Elektronenmikroskopie
} Institut fÃ¼r Botanik (210)
} UniversitÃ¤t Hohenheim
} Postfach, 70593 Stuttgart
} Tel. 0711-45922180
} Fax. 0711-45923355
}
}
} ==============================Original Headers==============================
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--

J. Faerber
IPCMS-DSI
Institut de Physique et Chimie des Matériaux de Strasbourg
Département Surfaces et Interfaces
23, rue de Loess ; BP43
67034 Strasbourg CEDEX 2
France

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

==============================Original Headers==============================
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Hello All,
I have just sent a copy of original Kodak formula for D-19 to Anne. It
was copied from over 40 years old Kodak leaflet delivered with
autoradiography plates. If anyone is interested in I can provide a link
to PDF file with it. We have been using it over 30 years with 1+2
dilution (standard contrast) or 1+1 dilution (high contrast) for
Kodak 4489 or Agfa Scientia plan films exposed at 80 kV.

It slightly differs from Jacques formula for D-19b.

Best regards Oldrich

--
Oldrich Benada
Institute of Microbiology AS CR, v.v.i.
Videnska 1083
142 20 Prague 4
Czech Republic

tel.: 241062399

On Wed, 3 Sep 2014 03:00:30 -0500, jacques.faerber-at-ipcms.u-strasbg.fr
wrote :
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} ----------------------------------------------------------------------------
}
} Hi
}
} For those who are intersted (and have some time for that stuff), here
} is the recepee of the D19b :
}
} Grammes for 1l final solution. To be diluated at 30-40 Â°C in
} distilled water to make a concentrated mother solution at 1/4 or 1/3.
} This mother solution will be kept in an air and light free bottle at
} { 18Â°C and diutated as wanted for use.
}
} GÃ©nol (MÃ©tol, Rhodol, Elon) 2.2
} Sulfite de Sodium 72
} Hydroquinone 8.8
} Carbonate de Sodium 48
} Bromure de potassium 4
}
} Sorry, it's with french names, but it should be easy to find the
} translations.
}
} Hoping it can help someone !
}
} Jacques
}
} --
}
} J. Faerber
} IPCMS-DSI
} Institut de Physique et Chimie des MatÃ©riaux de Strasbourg
} DÃ©partement Surfaces et Interfaces
} 23, rue de Loess ; BP43
} 67034 Strasbourg CEDEX 2
} France
}
} Tel 00 33(0)3 88 10 71 01
} Fax 00 33(0)3 88 10 72 48
} E-mail : Jacques.Faerber-at-ipcms.unistra.fr
}
}
}
}
}
}
} Le 02/09/2014 11:55, Anne.Heller-at-uni-hohenheim.de a Ã©crit :
} } ----------------------------------------------------------------------------
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} } America To Subscribe/Unsubscribe --
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7, 43 -- From benada-at-biomed.cas.cz Wed Sep 3 04:42:06 2014
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7, 43 -- From: Oldrich Benada {benada-at-biomed.cas.cz}
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From ueyqquje-at-kanzlei-schmoelz.at Wed Sep 3 12:02:16 2014
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Message-ID: {1459F2F4.9337952-at-kanzlei-schmoelz.at}

Hello Phill, hello All,
The link is:

{http://www2.biomed.cas.cz/~benada/D-19.pdf}

My best regards

Oldrich

--
Oldrich Benada
Institute of Microbiology AS CR, v.v.i.
Videnska 1083
142 20 Prague 4
Czech Republic

On Wed, 3 Sep 2014 09:22:32 -0400, Philip Oshel {oshel1pe-at-cmich.edu}
wrote :
} Oldrich,
}
} I would appreciate a link to this pdf, please.
} Thanks.
}
} Phil
}
} On 09/03/2014 05:57 , benada-at-biomed.cas.cz wrote:
} }
} } Hello All,
} } I have just sent a copy of original Kodak formula for D-19 to Anne.
} } It was copied from over 40 years old Kodak leaflet delivered with
} } autoradiography plates. If anyone is interested in I can provide a
} } link to PDF file with it. We have been using it over 30 years with
} } 1+2 dilution (standard contrast) or 1+1 dilution (high contrast) for
} } Kodak 4489 or Agfa Scientia plan films exposed at 80 kV.
} }
} } It slightly differs from Jacques formula for D-19b.
} }
} } Best regards Oldrich
} }
}
} --
} Philip Oshel
} Microscopy Facility Supervisor
} Biology Department
} 024C Brooks Hall
} Central Michigan University
} Mt. Pleasant, MI 48859
} (989) 774-3576

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Hi Oldrich and all

Thanks for the link.

I asked me from time to time why the "b" on the name D19-b. What about
the "D19-a" and here we have the answer.

But the differences between both are smale. They have the same
genol/hydroquinone ratio, so the same developping caracteristic on grey
levels and contrast rendering. As the D19 has less accelerator, more
preserving agent and less "anti-voile" (what's the name ine english ?),
it gives probably a less agressive developpement and is more tolerant on
developping time variations.

In comparison the D76, which is for fine grain and low contraste, has 2g
genol, 5g hydroquinone, 100 g sodium sulfite and no sodium carbonate.

Probably the evolution to the "b" version was made to gain some contrast
and/or to need a shorter developping time.

Best regards

Jacques

Le 03/09/2014 19:47, benada-at-biomed.cas.cz a écrit :
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
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} ----------------------------------------------------------------------------
}
} Hello Phill, hello All,
} The link is:
}
} {http://www2.biomed.cas.cz/~benada/D-19.pdf}
}
} My best regards
}
} Oldrich
}

--

J. Faerber
IPCMS-DSI
Institut de Physique et Chimie des Matériaux de Strasbourg
Département Surfaces et Interfaces
23, rue de Loess ; BP43
67034 Strasbourg CEDEX 2
France

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

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Hello Listservers,
Is there anyone out there that still burns objective aperture strips for
their TEMs? I plan to try this with a Denton DV 502A evaporator and an old
Hitachi 7600 strip (not thin foil).
My questions are: At what vacuum do you burn the aperture? How long do you
burn the aperture and at what current?

Thank you,
Michael Delannoy

==============================Original Headers==============================
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Hi Michael

The best way to heat up your aperture strip, is to mount it so that the
aperture area rests between two pieces of platinum. Why this method, well
using a platinum boat does not usually provide an even level of heat along
the aperture area.

Increase the current slowly and the aperture itself will start to glow.
Continue until the aperture is an orange-red, stop, and watch the dark
contamination around the apertures gradually disappear. Many people will
suggest white heat, but in my experience this has a high possibility of
damaging the aperture shape after one or two cleaning sessions. Orange-red
is enough to obtain a clean aperture, and this level of heat will work for a
number of cleaning procedures before any noticeable distortion of the
apertures themselves.

The current drawn will vary on the contact between the aperture and the
platinum supports. Slowly increasing the current will ensure you do not
supply too much current and damage the aperture strip.

Good luck

Steve

Steve Chapman FRMS
Protrain for Consultancy and Courses in Electron Microscopy
Mobile +44 (0) 7711 606967 Fax +44 (0)1280 814007
www.emcourses.com

-----Original Message-----
X-from: mdelann1-at-jhmi.edu [mailto:mdelann1-at-jhmi.edu]
Sent: 04 September 2014 14:49
To: protrain-at-emcourses.com

Michael,

The higher the vacuum the better, but at least 10^-6. I just go to
bright red hot or orangish then let it cool.

Phil

} Hello Listservers,
} Is there anyone out there that still burns objective aperture strips for
} their TEMs? I plan to try this with a Denton DV 502A evaporator and an old
} Hitachi 7600 strip (not thin foil).
} My questions are: At what vacuum do you burn the aperture? How long do you
} burn the aperture and at what current?
}
} Thank you,
} Michael Delannoy
}
--
Philip Oshel
Microscopy Facility Supervisor
Biology Department
024C Brooks Hall
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576

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Greetings,

Hooke College of Applied Sciences, located in Westmont, IL, is offering a Transmission Electron Microscopy short course October 7-9, 2014. In addition to lectures, this course emphasizes hands-on training using state-of-the-art equipment.

For further TEM training details and registration information, please follow the link below:

http://www.hookecollege.com/courses/course.asp?COURSE_ID=53

Best regards-
__________________________________________________
Chris Gorman
Admissions Specialist
Hooke College of Applied Sciences
850 Pasquinelli Drive
Westmont, IL 60559-5539 USA
630-887-7100 (tel)
630-887-7412(fax)

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Group,

Would like to hear from academic science departments on the subject of instrument maintenance.

How are different classes of instruments handled? i.e. computers, optical microscopes, small lab tools like PH meters and ovens, and major lab tools SEM, Spectrometers

Do you have a dedicated person to perform maintenance or is it done by lab personnel?

Do you use contract support or internal university resources?

Is maintenance a line item in your operating budget? How is that calculated?

Thanks for your feedback

Roy Beavers
rbeavers-at-smu.edu

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Email: k.jack-at-uq.edu.au
Name: Kevin Jack

Organization: The University of Queensland, Australia

Title-Subject: [Filtered] Dual Beam SEM/FIB Engineer at the Centre for Microscopy and Microanalysis,
The University of Queensland

Message: Please see the following details for a newly advertised position at the Centre for
Microscopy and Microanalysis, The University of Queensland, Australia.

Full details of the position and the application process can be found at
http://jobs.uq.edu.au/caw/en/job/496351/dual-beam-sem-engineer

The Centre for Microscopy and Microanalysis (CMM) has a world standard laboratory equipped with
state-of-the-art instrumentation and techniques. The Centre has nineteen electron microscopes, two
XRD units, one XPS unit and provides leading edge capability in micro structural analysis to the
staff and students across a broad range of disciplines at the University of Queensland. In addition,
the CMM provides training to clients in relevant techniques and in solving characterization problems
in a wide range of industrial, environmental and biological processes.

The role:

This role is to manage and operate a dual-beam scanning electron microscope which will be located in
the Centre for Microscopy and Microanalysis (CMM). This role involves a number of aspects including
the training of new users in the proper and effective usage of the instrumentation, in the
preparation of sample specimens for transmission electron microscopy; the production of high quality
3-dimensional (tomographic) images the measurement of analytical data and crystallographic data
from freshly milled samples. Moreover, aiding clients in experimental design and in the
interpretation of high quality data will be a highly significant aspect this management position.

The person:

Applicants should possess qualifications and/or experience in dual beam scanning electron microscopy
and transmission electron microscopy and have the demonstrated ability to manage the technical
operations and maintenance of the FIB/SEM and be committed to understanding and servicing the
requirements of its user community. The applicant will be an enthusiastic and capable individual
with excellent oral and written communication skills and be willing to take responsibility for the
facility and to facilitate the research programs of the users.

Full details of the position and the application process can be found at
http://jobs.uq.edu.au/caw/en/job/496351/dual-beam-sem-engineer

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Hi, Roy

Great question.

Two quick thoughts:
Just a reminder that MME offers customized maintenance courses for microscopy.

Also, I had a intriguing meeting with Judith LaCoste recently, at M&M, and we discussed the whole process of aligning and calibrating microscopes. It is worth having a discussion with her. You can reach her at jlacoste-at-miacellavie.com.

CAVEAT:
MME offers customized on-site courses on a for-fee basis.

Best regards,
Barbara Foster, President & Chief Consultant
Microscopy/Microscopy Education*
www.MicroscopyEducation.com

*A subsidiary of The Microscopy & Imaging Place, Inc.
7101 Royal Glen Trail, Suite A
McKinney, TX 75070
P: 972-924-5310
F: 214-592-0277

MME is currently scheduling courses for now and through January 2015. Call us today for a free training evaluation.

At 09:07 AM 9/5/2014, rbeavers-at-mail.smu.edu wrote:

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

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Hi Barbara

We run a 5 day "Monitoring & Maintaining the Electron Microscope" Course
which has been extremely successful around the world including the USA. By
the word "microscopy" I assume you are talking about light microscopes?

Regards

Steve

Steve Chapman FRMS
Protrain for Consultancy and Courses in Electron Microscopy
Mobile +44 (0) 7711 606967 Fax +44 (0)1280 814007
www.emcourses.com

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From pfujupae-at-ertelecom.ru Sat Sep 6 08:23:25 2014
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Dear Listers,

At UMD NISP Lab we are trying to restore and return back to operation
diffusion-pumped Pelco PAC-1 Advanced Coater 9500, I am wondering if
someone may have a copy of the manual that (s)he would be willing to share.

PDF is preferred, but I would pay costs of copying and mailing if that
is the only way.

Thank you very much beforehand,
--
Valery Ray - also with NISP, UMDCP Nanocenter
=============================================
PBS&T, MEO Engineering Co., Inc.
290 Broadway, Suite 298
Methuen, MA 01844 USA
Phone: +1-978-296-5063 - leave a message with call-back number
US Mobile: +1-978-305-0479
Skype: pbstmeo
E-Mail: vray-at-partbeamsystech.com
UMD E-Mail: vray-at-umd.edu
Web: www.partbeamsystech.com
Web: www.freudlabs.com

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4, 35 -- Date: Sun, 07 Sep 2014 17:59:07 -0400
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Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

I was trying to resurrect 20 y.o. W - filament LEO 1455 VP-SEM and
stumbled into apparent software problem: during the start-up system
would progress to "Initializing EO DLL" and stop at 25% of the progress
bar. There are no error messages, dongle is installed on parallel port,
and fiber optic communication between PC and the tool seem to be
operational.

Wondering if there are users of this instrument who have seen similar
problem and may have any ideas. Any kind of manuals, either user-level
or service/maintenance would also be appreciated, as after the repair
installing filament and aligning the column would be the next adventure...

Thank you very much beforehand,
--
Valery Ray - also with NISP, UMDCP Nanocenter
=============================================
PBS&T, MEO Engineering Co., Inc.
290 Broadway, Suite 298
Methuen, MA 01844 USA
Phone: +1-978-296-5063 - leave a message with call-back number
US Mobile: +1-978-305-0479
Skype: pbstmeo
E-Mail: vray-at-partbeamsystech.com
UMD E-Mail: vray-at-umd.edu
Web: www.partbeamsystech.com
Web: www.freudlabs.com

==============================Original Headers==============================
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From tujyma-at-weekendcruiser.com Mon Sep 8 05:48:52 2014
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Hello Listservers,

Our EDAX Genesis system needs an update to windows 7.

The current offer from the supplier includes an unexpecedly expensive PC (} 3000 Euros). I'm wondering if there are alternatives to this scenario.

Is there anyone out with experience on this topic?

Best regards

Bettina

________________________________________________________________
Â
Dipl. Ing. (FH) Bettina Wolpensinger
Â
Abteilung Photovoltaik
Photovoltaics Department
Â
Elektronenmikroskopie
Electron Microscopy
Institut fÃ¼r Solarenergieforschung GmbH
Am Ohrberg 1
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(BÃ¼ro) oder 218 (Mikroskop)
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Vorsitzender des Aufsichtsrats: Ministerialrat Dr. Hans Schroeder
________________________________________________________________
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From pharmacy_reasonable9-at-telecolumbus.net Mon Sep 8 11:17:18 2014
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Email: rnichols-at-bcm.edu
Name: ralph nichols

Organization: Baylor College of Medicine

Title-Subject: [Filtered] Giudelines for TEM Digital Camera Use

Message: Hello Listers,

I hope all had a great summer. I'm about to get a digital camera for my
TEM (YES!). This is a CORE facility. Some
of our researchers have shied away from using it because of the film
imaging. I anticipate more of them will be wanting to use the TEM. My
questions for those whom manage TEM with digital cameras with multiple
users: what kind of guidelines that I need to use in managing the camera
such as should I
restrict the kind of flash drives to down load the images and any other
suggestions? Please email me directly to rnichols-at-bcm.edu.

Thanks

Ralph Nichols

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Contents Retrieved from Microscopy Listserver Archives
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Dear List Members,

Following a recent upgrade of our cryo-TEM instruments, we are exploring the possibility of selling some of the auxiliary equipment previously serving the Tecnai F20 TEM (FEI company). All items detailed are performing (beyond) specifications and are in perfect working conditions.

The auxiliary equipment list includes:

· 1. One GIF 2002 Gatan’s 2nd generation post-column energy filter, equipped with cryo-sensitive 2K x 2K MSC CCD camera, controllers and a retractable TV-rate camera. The system has always been under service contract.

· 2. Two DH626 Single Tilt Liquid Nitrogen Cryo Transfer Holders, 60º and 70º, with transfer stations, Gatan Inc

· 3. One CT3500TR Single Tilt Rotation Liquid Nitrogen Cryo Transfer Holder, Transfer Station and Controller, Gatan Inc.

· 4. Two Advanced-Tomography Holders Model 2020, Fischione Instruments, Inc.

· 5. One: CompuStage Rotation Holder, Room Temperature, FEI company

Pictures: www.dropbox.com/sh/eaa00cb3wuleg2o/AAD2GPawJUsRNNPxRppy5sq4a

If you are interested in any of the above items please contact me at dorit-at-burnham.org for more information and pricing.

Best wishes

dorit

__________________________

Dorit Hanein, PhD
Professor
Bioinformatics and Structural Systems Biology Program
Sanford Burnham Medical Research Institute

==============================Original Headers==============================
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Contents Retrieved from Microscopy Listserver Archives
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AFMWorkshop's free livestreaming AFM Seminar Series continues tomorrow
with a discussion led by Paul West, Ph.D., on AFM Probe-Cantilever
Selection.

AFM operators must choose from a wide array of probe types when
scanning samples. This seminar will cover the probe-cantilever types
that work best for various types of samples and modes.

To Register, visit AFMWorkshop's UStream Channel:
http://www.ustream.tv/channel/Atomic-Force-Microscopes.
All participants are encouraged to ask questions live.

Thank you,
Pamela Stone
AFMWorkshop, Inc.

Pamela K. Stone
AFMWorkshop, Inc.
1434 E. 33rd Street
Signal Hill, CA 90755
(888)671-5539 (O)
(562)274-2788 (M)
www.afmworkshop.com

==============================Original Headers==============================
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Dear Listers,

Just a reminder about tomorrow's webinar:A Better Choice than Photoshop for Color Brightfield Microscopy

Dear All,

thank you for all who responded to my question. In the meantime I
tested the substitute for Kodak D-19 from Ted Pello for 1 gallon. It
gives 1 gallon stock solution. For Kodak plates 4489 4 min in 1+2
dilution the result is fine. It is a really a substitute for Kodak D-19.

Best regards,
Anne
Dr. Anne Heller
AG Elektronenmikroskopie
Institut fÃ¼r Botanik (210)
UniversitÃ¤t Hohenheim
Postfach, 70593 Stuttgart
Tel. 0711-45922180
Fax. 0711-45923355

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Email: jhyun-at-gatan.com
Name: John Hyun

Organization: Gatan

Title-Subject: [Filtered] EELS & EFTEM Analysis Training School

Message: EELS & EFTEM Analysis Training School
Gatan, Inc. Pleasanton, California, USA
October 14-17, 2014

Want to refresh or advance your EELS and EFTEM knowledge? Join us for an
intensive 4-day training school that incorporates lectures, computer
laboratories, and microscope practicals to provide participants with
comprehensive, hands-on training on key EELS and EFTEM topics and
technology.

Online registration is now open:
http://ftp.gatan.com/training/formOct2014.php. School fee: $1,750.00 US
Dollars. Space is limited.

Overview: Transmission electron microscopy (TEM) reveals details of
natural and man-made structures at the micrometer, nanometer, and even
sub-nanometer scale. Energy-filtered TEM (EFTEM) and electron
energy-loss spectroscopy (EELS) are the ideal analytical partners to the
high spatial resolution provided by TEM in both the conventional and
scanned (STEM) imaging modes.

This course reviews the basic theory and practice of EELS imaging and
analysis in the TEM, but its main emphasis is on practical techniques,
optimum deployment of Gatan hardware and software systems, and advanced
EELS and EFTEM applications. Some prior experience with EELS, EFTEM, and
Gatan systems is recommended, as is a good familiarity with TEM/STEM
instrumentation and techniques. By the end of the course, participants
can expect to know how best to optimize the performance of their Gatan
EELS hardware as well as their EELS and EFTEM experimental setups in
order to capture and extract the maximum amount of information from
their TEM samples.

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Email: twh-at-cen.dtu.dk
Name: Thomas

Organization: DTU Cen

Title-Subject: [Filtered] Optical bench software

Message: Hi.

We just started up a course in electron microscopy (both TEM/STEM/SEM
etc). For illustrating the effects of condenser lenses and apertures on
the final probe in the SEM, it would be nice to have some optical bench
software. Does anyone know of such software? Preferably freeware, but it
doesn't have to be.

Thanks.

Thomas

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Email: Kirsty.MacLellan-Gibson-at-nibsc.org
Name: Kirsty MacLellan-Gibson

Organization: NIBSC

Title-Subject: [Filtered] cryo-workshop 2014

Message: NIBSC with JEOL (UK) Ltd. LEICA and Gatan are holding a
dedicated cryo-workshop based on technologies installed at the NIBSC
Imaging laboratory, Potters Bar, London, UK. The course will be held
over 5 days commencing Monday 27th October through to Friday 31st
October 2014. Delegate numbers will be strictly limited to ensure
maximum hands-on training.
The course will embrace cryo-preparation techniques using Cryo-FEGSEM,
Cryo-TEM with GIF quantum and many associated preparation techniques:
High pressure freezing (HMP010), Freeze fracture (Leica/BAF060),
Cryo-sectioning, Freeze substitution, Vitreous thin films (plunge
freezing), Tomography and High resolution Cryo-FEG SEM. This year new
automated acquisition solutions will also be taught.
The ethos of the course is very much Â“hands-onÂ” with technical support
lectures from scientists in these fields and application support from
JEOL, LEICA and invited specialists from University of Lausanne and Max
Plank Institute.

The course cost is Â£550.00 including conference dinner and lunches but
excluding accommodation.

Applications are invited, please include a brief description of your
current interests to Gill Cathro Email: Gill.Cathro-at-nibsc.org; Tel: +44
(0) 1707 641561; Fax: +44 (0) 1707 641578

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From saetyqeu-at-ramnet.su Wed Sep 10 15:18:43 2014
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Dear Colleagues,

WE are in the process of searching for alternative systems to replace our
aging Toshiba UPS 1600XPs, we use them to backup our transmission electron
microscopes. Can anyone suggest a comparable system that works for you?

Thank you very much in advance.

Xinran

________________________________________

Xinran Nick Liu, M.D. & Ph.D.
Director of CCMI Electron Microscopy Core Facility
Research Scientist in Cell Biology
Yale University School of Medicine
333 Cedar Street, SHM I-E16A
New Haven, CT 06520
Office: 203.785.4050
Lab: 203.785.5390
http://medicine.yale.edu/ccmi/em

==============================Original Headers==============================
10, 35 -- From xinran.liu-at-yale.edu Thu Sep 11 10:21:38 2014
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10, 35 -- From: "Liu, Xinran" {xinran.liu-at-yale.edu}
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From affordable_drugstore4-at-arcotoys.com Thu Sep 11 12:02:50 2014
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Dear Xinran,
At a past job, I tested and subsequently had installed UPS/power
conditioning solutions from PowerVar. I found them to have good prices,
small units, and a range of battery capacity options (we were looking
protection against momentary power loss or for just a few minutes of run
time for SEM's and FIB's, to allow for gracious shut down.) The units worked
well and did in fact come into action more than once.
Good luck!

-----Original Message-----
X-from: xinran.liu-at-yale.edu [mailto:xinran.liu-at-yale.edu]
Sent: Thursday, September 11, 2014 11:37 AM
To: LES-at-ZSGENETICS.COM

Dear Colleagues,

WE are in the process of searching for alternative systems to replace our
aging Toshiba UPS 1600XPs, we use them to backup our transmission electron
microscopes. Can anyone suggest a comparable system that works for you?

Thank you very much in advance.

Xinran

________________________________________

Xinran Nick Liu, M.D. & Ph.D.
Director of CCMI Electron Microscopy Core Facility
Research Scientist in Cell Biology
Yale University School of Medicine
333 Cedar Street, SHM I-E16A
New Haven, CT 06520
Office: 203.785.4050
Lab: 203.785.5390
http://medicine.yale.edu/ccmi/em

==============================Original Headers==============================
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Title-Subject: [Filtered] Service charge for TEM work

Message: Dear Listserver
I would like to know per hour charge to (1) use an outside
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Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

The University of Illinois Materials Research Lab is sponsoring its 3rd Annual Biostructures Conference.

New this year are student presentation and poster competitions.

One does not have to compete to have an informative time—two days of invited experts will be presenting engaging sessions:

Dr. Bill Wilson (Materials Research Lab): Devices to make proteins on the nano scale
Dr. David Hall (Albert Einstein College of Medicine): High tech solutions for Biological samples/C. elegans.
Dr. Sivaguru Mayandi (Institute for Genomic Biology): Confocal and the 3D shape of pollen.
Dr. Adam Stern ( UI Veterinary Medicine): The art of histology.
Linda Arseneau (UI Division of Research Safety): Biological safety
Dr. James Nardi (UI Entomology): Preparing insects for microscopy
Dr. Scott MacLaren (Materials Research Lab): Biological AFM
Lou Ann Miller (Materials Research Lab): Biological TEM
Dr. Stephan Lezmi (UI Veterinary Medicine): Immunohistochemistry
JPK (a company that will demo instruments and speak): Biological AFM
Mark Reitsma (Oxford-Asylum, a company that will demo instruments and speak): Biological AFM

The 2-day workshop includes all presentations, facility tours, student competitions, a vendor fair and plenty of tasty food during both days!

If you are interested in competing, please see the rules online. You can enter the completion when you register for the workshop online.

Workshop information, student presentation and poster competition rules, directions to MRL and registration are online at: www.mrl.illinois.edu/biostructures2014

Participant Registration: $60
($20 of the fee will be refunded to students selected to present.)

Exhibitor Registration: $300

Hope we see some of you here!
Lou Ann Miller

{ { { { { { { { { { { { { { { { {} } } } } } } } } } } } } } } }
Lou Ann Miller
Biological Electron Microscopy
Frederick Seitz Materials Research Laboratory
Lab Room 125 Office 374 MRL
104 South Goodwin Ave
Urbana, IL 61801
Campus mail code: MC-230

217-244-1567
lamiller-at-illinois.edu
http://mrl.illinois.edu

==============================Original Headers==============================
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Name: Chya Yan

Organization: Northwestern University

Title-Subject: [Filtered] Osmium Tetroxide waste disposal procedures

Message: Dear Listserver,

I have a question regarding to the disposal procedure for Osmium
Tetroxide. I am new to TEM and have little knowledge of the staining
agent OsO4. According the procedure I found online, I added twice amount
of corn oil in the OsO4 bottle to neutralize it. However, after several
hours, I observed the solution separated with the oil on the top while a
black layer on the bottom. Should I stir the solution then? Since this
is my first time dealing with the waste, I don't know what should I
observe to know the neutralization process is done. Could someone
explain more details of the procedure? I really appreciate your help!

Chya Yan

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Name: Randy

Title-Subject: [Filtered] SEM for Red Blood Cells

Message: Hi there,

I'm trying to image red blood cells in SEM. Are there good protocols
that are simple for novice sample preparers? We are looking especially
for things that work using easy to get reagents (no exotic, expensive,
or highly toxic chemicals). We're having a lot of trouble, any
suggestions will be greatly appreciated!

Thanks!

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From mohmedi-at-coltel.ru Sat Sep 13 10:13:23 2014
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Hi Michael, just burn your aperture plate by holding it in the blue part of
a gas flame by means of forceps. Shape of the hole does'nt matter much.
Success Peter

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3, 25 -- Subject: TEM cleaning apertures
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If the aperture is Molybdenum, then as alternative to burning you can
soak it overnight in warm (30C to 40C, candle warmer works perfectly
here) solution of 80% Micro-90 in 20% DI water, followed by 20min
ultrasonic cleaning next morning in the solution of 5% Micro-90 95% DI
water, followed by dipping in 99.9% IPA "low residue analytical grade
for chromatography," followed by blow-dry by CDA, or DN2, or
clean-room-approved duster. This procedure also works for anodes and
beam-limiting apertures of SEM, if they are made of Mo.

Valery Ray - also with NISP, UMDCP Nanocenter
=============================================
PBS&T, MEO Engineering Co., Inc.
290 Broadway, Suite 298
Methuen, MA 01844 USA
Phone: +1-978-296-5063 - leave a message with call-back number
US Mobile: +1-978-305-0479
Skype: pbstmeo
E-Mail: vray-at-partbeamsystech.com
UMD E-Mail: vray-at-umd.edu
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Web: www.freudlabs.com

On 9/14/2014 5:01 PM, p.elbers90-at-upcmail.nl wrote:
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} Hi Michael, just burn your aperture plate by holding it in the blue part of
} a gas flame by means of forceps. Shape of the hole does'nt matter much.
} Success Peter
}
}
}
} ==============================Original Headers==============================
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} 3, 25 -- Subject: TEM cleaning apertures
} 3, 25 -- From: Peter Elbers {p.elbers90-at-upcmail.nl}
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Name: hadeel

Title-Subject: [Filtered] Asphalt under sem

Message: hi ,can i study Asphalt under the scanning electron microscope in high vacuum mode and how
they are
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It seems a little strange that you would ask the microscopy community how asphalt is being studied. I assume that you would have a question that you would like to answer rather than asking about others questions and techniques.

Thin layers of asphalt on aggregate might be studied at high vacuum. You would need to take normal precautions for dealing with insulating samples, i.e., low voltage and/or a metal coating.

You might also be prepared for some contamination in the chamber. You might volatilize some organic components that would condense within your chamber leading to a need for more frequent cleaning.

We tried looking at a polished section of asphalt in a high vacuum SEM years ago. We had problems getting to operational vacuum. Between volatile components and porosity, the sample would not pump down in the allowed time. We ended up buying an SEM with variable pressure capability. We did not have to pump to such a high level so we could reach operational vacuum quicker. VP mode meant we did not have to contend with charging. Also, we were in a vacuum mode where the residual gas helped sweep away the components that came off the sample. We did not have to worry about them depositing in the chamber.

I recommend that you locate a VP-SEM if you plan to do much asphalt or concrete work.

Warren Straszheim
Materials Analysis and Research Lab
Iowa State University

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Organization: Queen's University

Title-Subject: [Filtered] how to choose between different Wehnelt Apertures?

Message: Hi there, I am changing filament for Phillip CM20 TEM, and just found we have several
different Wehnelt Apertures with various hole sizes in stock. So, what's the difference for
different sizes of the apertures? How to choose one?

Cheers,

Jason

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Hello Jason,
For Philips CM series following Wehnelt diaphragm are recommended.

Standard Tungsten Filament: 0.5 mm
LaB6 Filament: 0.3 mm

Best regards Oldrich

--
Oldrich Benada
Institute of Microbiology AS CR, v.v.i.
Videnska 1083
142 20 Prague 4
Czech Republic

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7, 39 -- Apertures?
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McCrone Associates, Inc., a Chicago-area analytical and research laboratory, is looking to expand our staff by seeking several highly motivated and curious research scientists to solve materials and contamination problems using modern microscopy and microanalysis for our clients in a variety of industries. These positions offer a significant amount of autonomy and require the skills to handle multiple projects at any given time.

If you are an experienced microanalyst and looking for an interesting career in materials analysis, in a modern and innovative laboratory, please browse our current job opportunities at www.mccrone.com/careers.

You may apply directly through the website or by submitting a resume and cover letter to Gina Schuster, Director of Human Resources at gschuster-at-mccrone.com.

Elaine F. Schumacher | Senior Research Scientist
McCrone Associates, Inc.
850 Pasquinelli Drive . Westmont, IL 60559
P. 630.887.7100 | F. 630.887.7417 | www.mccrone.com

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Listers,
Anyone out there know the model number of the Haskris chiller that
shipped with the JEOL 840A SEM? Please reply offline. Thanks.

Gary M. Easton
Scanners Corporation
90 Aileron Court, Ste. 6
Westminster, Maryland USA 21157
410.857.7633 (Office)
443-524-9631(Fax)
717.634.2226 (Mobile)

==============================Original Headers==============================
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9, 19 -- Subject: JEOL 840A SEM Chiller
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Email: susan.vanhorn-at-stonybrook.edu
Name: Susan C. Van Horn

Organization: Stony Brook Univ

Title-Subject: [Filtered] tissue fixed in glutaraldehyde

Message: Has anyone experienced difficulty in sectioning (thick or thin) tissue that has been fixed
in glutaraldehyde only and let sit in fix for days, weeks or even months??? embedding or sectioning
problems??
thanks
Sue

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From botgibya-at-smart.az Wed Sep 17 03:25:11 2014
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Listers,
Thanks everyone who responded to my request, I appreciate the help.

Gary

Gary M. Easton
Scanners Corporation
90 Aileron Court, Ste. 6
Westminster, Maryland USA 21157
410.857.7633 (Office)
443-524-9631(Fax)
717.634.2226 (Mobile)

==============================Original Headers==============================
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***************************************************************************************
Forwarded from "Ask a Microscopist"
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realname - Emilie Colton
Email - e.colton-at-utoronto.ca
ORGANIZATION - University of Toronto
EDUCATION - Graduate College
LOCATION - Mississauga, Ontario, Canada
SUBJECT_OF_QUESTION - Emerging Market Opportunities and Existing
Limitations of Super Resolution Light Microscopy
QUESTION - Hello,

I am a graduate student at the University of Toronto, and I am
conducting research on the emerging market opportunities and the
existing limitations surrounding super resolution light microscopy. As
per our instructions for this project, we are required to inquire
expert's opinions and insight on the matter. We would greatly appreciate
any information/input so as to develop a detailed understanding of the
subject.

Thank you in advance,
Emilie Colton

==============================Original Headers==============================
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Email: kkusser-at-vgtifl.org
Name: Kim

Organization: VGTI-FL

Title-Subject: [Filtered] Hypoxia setup on an inverted microscope

Message: Hi all,

We have a researcher who would like to do long term imaging of cardiomyocyt=
es under hypoxic conditions.

Our microscope is currently setup with a CO2 chamber. (DV Elite on a Olympu=
s inverted stand).

My idea is that we could add nitrogen (via a gas cylinder)... throw a flow =
meter on it ( similar to the one we have for the CO2). ....run the N2 line =
to the CO2 enclosure. Then, we would have to get some type of O2 sensor to =
dial in the correct mix of N2 and CO2 for the target %O2.

We would like to do this as cheaply as possible, and I was wondering if any=
one out there has any experience they would like to impart as to the feasib=
ility of my semi-formed idea.

Thanks

Kim

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From niepuli-at-jardine-motors.com Thu Sep 18 08:36:57 2014
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Message-ID: {5E040084.3389793-at-jardine-motors.com}

Dear Emilie,

I would be happy to discuss this project with you. Please contact me off-line at the phone number/email below.

Also, as an invited contributor, I wrote an overview article for BioPhotonics several years ago. Here is the link: http://microscopyeducation.com/images/Superresolution_BioPh_01_2011.pdf

Best regards,
Barbara Foster, President & Chief Consultant
Microscopy/Microscopy Education*
www.MicroscopyEducation.com

*A subsidiary of The Microscopy & Imaging Place, Inc.
7101 Royal Glen Trail, Suite A
McKinney, TX 75070
P: 972-924-5310
F: 214-592-0277

MME is currently scheduling courses for now and through Jan 2015. Call us today for a free training evaluation.

At 03:48 PM 9/17/2014, oshel1pe-at-cmich.edu wrote:

} ----------------------------------------------------------------------------
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Hello Dear Listers

The high tension cable in our Jeol 5600 LV is not working and would like to
replace it. Anybody having positive experience with a company that can do
this with low cost, in the USA or in Europe?
Thanks!
yorgos

Dr Yorgos Nikas
Athens Innovative Microscopy
Skra 36 Voula 16673 GREECE

Tel/fax +30 210 8957677
mobile +30 6945 107477
www.eikonika.net www.aim.cat
************************************

==============================Original Headers==============================
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5, 22 -- Subject: HT cable to replace
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Hi Yorgos

Servicing microscope for many years in Europe, I would talk to organisations
that service hospital equipment like x-ray machines. In those days it was
possible for these people to remake the cable if I provided the connections.

Good luck

Steve

Steve Chapman FRMS
Protrain for Consultancy and Courses in Electron Microscopy
Mobile +44 (0) 7711 606967 Fax +44 (0)1280 814007
www.emcourses.com

-----Original Message-----
X-from: eikonika-at-otenet.gr [mailto:eikonika-at-otenet.gr]
Sent: 19 September 2014 12:43
To: protrain-at-emcourses.com

Hello Steve
What about the high vacuum seal on the cap of the microscope. Does this
present a problem for the replacement of the cable?
With many thanks
yorgos

----- Original Message -----
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To: {eikonika-at-otenet.gr}
Sent: Friday, September 19, 2014 3:02 PM

X-from: robernal-at-u.northwestern.edu ()

This Question/Comment was submitted to the Microscopy Listserver
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Email: robernal-at-u.northwestern.edu
Name: Rodrigo Bernal

Organization: Northwestern University

Title-Subject: [Filtered] Grounding Electronics inside TEM

Message: I have a holder for electrical contacting which I am using to run an experiment in-situ
TEM. Parts of the circuit do go into the tip of the holder.

The circuit itself works pretty well in the TEM holder, whenever it is not inserted in the
microscope. However, when the holder is inserted, noise becomes really high. I have discovered that
I can connect the ground of my circuit to the microscope column (a screw or exposed metal in the
goniometer), and it reduces the noise back to the usual level. However, this action activates the
stage touch alarm, which locks the stage in place preventing any movement.

Does anybody have any details how the grounding in the TEM is done? Is the microscope exposed metal
connected to earth ground? Is there any point that one may use in the TEM for grounding without
affecting the stage?. Where does the TEM contacts the body of the holder electrically?

Thanks!!!

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From pharmacy-express1-at-seconduse.com Fri Sep 19 10:40:24 2014
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try these three:

http://www.dielectricsciences.com/index.html
http://www.parkermed.com/products
http://www.claymount.com/en/

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

On 9/19/2014 7:43 AM, eikonika-at-otenet.gr wrote:
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} Hello Dear Listers
}
} The high tension cable in our Jeol 5600 LV is not working and would like to
} replace it. Anybody having positive experience with a company that can do
} this with low cost, in the USA or in Europe?
} Thanks!
} yorgos
}
} Dr Yorgos Nikas
} Athens Innovative Microscopy
} Skra 36 Voula 16673 GREECE
}
} Tel/fax +30 210 8957677
} mobile +30 6945 107477
} www.eikonika.net www.aim.cat
} ************************************
}
}
} ==============================Original Headers==============================
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Contents Retrieved from Microscopy Listserver Archives
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I know the topic of service providers for electron microscopes comes up fai=
rly regularly, but what about for optical microscopes? Does anyone have exp=
erience or informed opinions about working with a third-party distributor o=
r service provider vs the manufacturer's service center?

We have a couple of Zeiss AxioImagers and an AxioZoom, and most of the time=
they just work. Once in a while a motorized component goes out. We have be=
en working with a distributor for service, but I wonder if there are benefi=
ts to outweigh the higher cost of working with the Zeiss service center.

None of us here have a lot of experience working on optical microscopes oth=
er than sitting in front on one to look at stuff, and the service guy isn't=
real strong on training. Are there some recommended guides for alignments =
or imaging conditions to use? We are looking at materials (thin films of me=
tals or polymers, silicon, particles and defects).

Thanks to everyone on this list who makes it such a valuable resource.

Jonathan Abbott

==============================Original Headers==============================
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11, 23 -- To: "Microscopy-at-microscopy.com" {Microscopy-at-microscopy.com}
11, 23 -- Subject: Optical microscope service
11, 23 -- Thread-Topic: Optical microscope service
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From myzedqa-at-comcastbusiness.net Fri Sep 19 13:39:54 2014
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The deadline for the student competition is approaching!

Win FAME.... GLORY..... $100 Cash Prize!

At our 3rd annual biological conference, we will be having 2 student competitions
for those working with biological substances, or a mix of biological and materials.

One competition is for oral presentation, one for poster presentations. You do not have to give a talk to enter a poster, but if you talk, we would like you to also present a poster.

How to enter:

1. Ask your advisor what credit card ( or account if in Illinois university system) to register on
( $60 for whole meeting, $20 refund if you present a talk) -- Price includes 2 days of food.

2. Write a 1/2 page abstract and gather any images you wish to submit.

3. Go to this web site, and look at the blue box menu to the right,
Choose payment type and begin the registration, there will be a button for
uploading files.

http://mrl.illinois.edu/events/conferences-workshops/third-annual-biological-imaging-workshop

Rules:

Poster:
http://mrl.illinois.edu/events/conferences-workshops/student-competition-poster-presentation

Presentation:
http://mrl.illinois.edu/events/conferences-workshops/student-competition-poster-presentation

I hope to see you and your work at the conference!

Lou Ann

{ { { { { { { { { { { { { { { { {} } } } } } } } } } } } } } } }
Lou Ann Miller
Biological Electron Microscopy
Frederick Seitz Materials Research Laboratory
Lab Room 125 Office 374 MRL
104 South Goodwin Ave
Urbana, IL 61801
Campus mail code: MC-230

217-244-1567
lamiller-at-illinois.edu
http://mrl.illinois.edu

==============================Original Headers==============================
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36, 37 -- Subject: Microscopy & analysis competition! Win Fame, Glory, Cash! at the
36, 37 -- University of Illinois.
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From kriuyyi-at-isaaceld.k12.az.us Tue Sep 23 13:17:34 2014
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Message-ID: {03F803F6.5889825-at-isaaceld.k12.az.us}

When I was at Intel in the SF Bay Area, we used two private optical shops with great satisfaction. They were:

Optotek
800-924-6023
http://www.optotek.net/

and

Serco
800-483-0508

John Mardinly, ASU

-----Original Message-----
X-from: jabbott-at-moxtek.com [mailto:jabbott-at-moxtek.com]
Sent: Friday, September 19, 2014 9:53 AM
To: John Mardinly

I know the topic of service providers for electron microscopes comes up fai= rly regularly, but what about for optical microscopes? Does anyone have exp= erience or informed opinions about working with a third-party distributor o= r service provider vs the manufacturer's service center?

We have a couple of Zeiss AxioImagers and an AxioZoom, and most of the time= they just work. Once in a while a motorized component goes out. We have be= en working with a distributor for service, but I wonder if there are benefi= ts to outweigh the higher cost of working with the Zeiss service center.

None of us here have a lot of experience working on optical microscopes oth= er than sitting in front on one to look at stuff, and the service guy isn't= real strong on training. Are there some recommended guides for alignments = or imaging conditions to use? We are looking at materials (thin films of me= tals or polymers, silicon, particles and defects).

Thanks to everyone on this list who makes it such a valuable resource.

Jonathan Abbott

==============================Original Headers==============================
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From medications-best2-at-triolan.net Tue Sep 23 15:08:12 2014
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Hello,

for our small lab we are looking for a used sputter coater to
coat SEM samples (Au, Pt or similar).
In particular, we would like to coat older decapsulated IC (integrated
circuit) models to get better results with voltage contrast imaging.
Since we're on a very small budget, I was wondering if anyone on this
list has an old sputter coater that is no longer in use and could be
sold to us.
Necessary repairs would be no problem as long as we can still get spare
parts.

Thank you,
Stefan

==============================Original Headers==============================
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Hello All,

A few months ago our lab acquired an old JEOL IC-848A SEM. We found
that the CRT is no longer working (the yoke coils are shorted out).
The CRT is a Toshiba, Model #E2755B7, with a Totoku yoke, Model
#EYS-03-0600.

We would like to replace this CRT. Online searches have shown one or
two places which have them, but those that do are without a yoke. Does
anyone know where to find a complete replacement CRT (or yoke)? Or, if
that's not possible, what a compatible replacement would be? Any help
would be appreciated.

Thanks you in advance,

Julius De Rojas

==============================Original Headers==============================
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5, 28 -- Subject: SEM JEOL IC-848A CRT (or yoke)
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Email: mclymer-at-datacolor.com
Name: Mark Clymer

Organization: Datacolor Inc.

Title-Subject: [Filtered] [LM] Webinar recording: "A Better Choice than
Photoshop..."

Message: The webinar recording of "A Better Choice than Photoshop for
Color Brightfield Microscope Images" is now available at
http://tinyurl.com/puyj339. In this webinar, Jerry Sedgewick (Imaging
and Analysis, LLC) demonstrates the common uses of Photoshop to adjust
color in scientific images, and then demonstrates better, faster results
using Datacolor ChromaCal.

CAVEAT: Datacolor is a manufacturer of color calibration solutions for
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Email: rms-at-angstrom.us
Name: Bob Sommerville

Organization: Angstrom Scientific Inc.

Title-Subject: [Filtered] Sales Representative Position Available, Mid
Atlantic

Message: Position: Technical Sales Representative - Mid Atlantic US
Territory: Dependent on the candidate but will include MD, DE, DC, VA
and Eastern PA.
Products: Sales and support of the following products/companies:
Hitachi: Bench-top Scanning Electron Microscopes (SEM)
DENSsolutions Heating and biasing holders for TEM
Kleindiek: Nano-manipulators & Stages
AppFive: Electron Diffraction (TEM)
Jordan Valley: X-ray Diffraction (XRD)
XEI: Plasma Cleaners
Deben Microscope Accessories
Microtrac Particle size measurement
MelBuild TEM sample holders

Position Description: This is an exciting position selling and
supporting Hitachi bench-top scanning electron microscopes which expand
on the capabilities of optical microscopes. In addition, Angstrom
Scientific Inc. represents a number of leading companies in the electron
microscopy and related markets and sells select used scanning electron
microscopes. The ideal candidate will preferably have a demonstrated
track record of success ( 2-3 years) selling technical products into the
academic, research and industrial markets. Optical microscopy or SEM/TEM
and/or particle measurement related experience is a definite plus.
Academic background should be at min an associateÂ’s degree in science
(BS or masters preferred) The successful candidate will live in the
above listed states.
Angstrom Scientific Inc is a growing distributor of products targeted to
the nanotechnology, research and semiconductor markets with exciting new
and novel technology
Angstrom Scientific Inc offers competitive salary and benefits,
commission, car allowance and expenses. We are an equal opportunity
employer.
Interested candidates should submit their resume together with salary
history to: Bob Sommerville, General Manager, Angstrom Scientific Inc.
e-mail: rms-at-angstrom.us

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Email: jcderojas-at-ucdavis.edu
Name: Julius De Rojas

Organization: UC Davis/Department of Physics

Title-Subject: [Filtered] JEOL IC-848A CRT Replacement

Message: Hello,

Our lab has recently acquired an old JEOL IC-848A that was out of
commission for two years. We've finally got it up and started, but we
found that we have a bad CRT. The yoke coils seems to be shorted out.

The CRT is a Toshiba, Model #E2755B7, with yoke made Totoku, Model
#EYS-03-0600. Does anyone know where a replacement yoke can be
purchased? Or, if not, what an equivalent replacement CRT would be?

Any assistance would be much appreciated.

Julius De Rojas

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From ruequsiu-at-garciastents.com Wed Sep 24 22:07:09 2014
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I am not that familiar with Jeol specifically, but done fair share of
upgrades and repairs.... does this monitor display regular RS-170 video
signal (analog TV standard?), or does it monitor a proprietary feed?

If monitor works with standard TV signal then any LCD TV monitor could
be plugged in instead; if signal is not standard then decent video input
card (like Epiphan VGADVI for example, or something slightly more
sophisticated in tough cases) could be used to digitize the signal and
display it on PC monitor.

Valery Ray - also with NISP, UMDCP Nanocenter
=============================================
PBS&T, MEO Engineering Co., Inc.
290 Broadway, Suite 298
Methuen, MA 01844 USA
Phone: +1-978-296-5063 - leave a message with call-back number
US Mobile: +1-978-305-0479
Skype: pbstmeo
E-Mail: vray-at-partbeamsystech.com
UMD E-Mail: vray-at-umd.edu
Web: www.partbeamsystech.com
Web: www.freudlabs.com

On 9/24/2014 4:11 PM, jcderojas-at-ucdavis.edu wrote:
} ----------------------------------------------------------------------------
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} Hello All,
}
} A few months ago our lab acquired an old JEOL IC-848A SEM. We found
} that the CRT is no longer working (the yoke coils are shorted out).
} The CRT is a Toshiba, Model #E2755B7, with a Totoku yoke, Model
} #EYS-03-0600.
}
} We would like to replace this CRT. Online searches have shown one or
} two places which have them, but those that do are without a yoke. Does
} anyone know where to find a complete replacement CRT (or yoke)? Or, if
} that's not possible, what a compatible replacement would be? Any help
} would be appreciated.
}
} Thanks you in advance,
}
} Julius De Rojas
}
} ==============================Original Headers==============================
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} 5, 28 -- Subject: SEM JEOL IC-848A CRT (or yoke)
} 5, 28 -- From: Julius De Rojas {jcderojas-at-ucdavis.edu}
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4, 41 -- Message-ID: {5424169D.3020405-at-partbeamsystech.com}
4, 41 -- Date: Thu, 25 Sep 2014 09:20:29 -0400
4, 41 -- From: Valery Ray {vray-at-partbeamsystech.com}
4, 41 -- Reply-To: vray-at-partbeamsystech.com
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4, 41 -- Subject: Re: [Microscopy] SEM JEOL IC-848A CRT (or yoke)
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realname - Megha Dubey
Email - meghandubey-at-gmail.com
EDUCATION - Graduate College
QUESTION - I am a PhD student, with thesis work completed, however with
an uncertainty of getting a PhD degree. I did my work in high
temperature alloy oxidation using different microscopic tools to obtain
information from macroscopic to atomic scale. I would like to develop my
career in field of microscopy. Are there any specialized course in USA
which will allow me to get a degree in field of microscopy?
Thank you for the help!

Regards,
Megha

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Organization: Lawrence Berkeley Laboratory

Title-Subject: [Filtered] Northern California Society for Microscopy
Fall Meeting

Message: The Northern California Society for Microscopy will hold its
fall meeting on Thursday October 9 at JH Technologies in Fremont, CA.

Please join us for dinner and an excellent group of speakers.
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Organization: Case Western Reserve University

Title-Subject: [Filtered] MSNO 2014 Fall meeting

Message: On behalf of Microscopy Society of Northeastern Ohio (MSNO),
I'd like to invite you to join MSNO Fall meeting.
This meeting is on Tuesday, October 14 at 4:00-8:30 pm at Goodyear
Auditorium, Goodyear Polymer Center, University of Akron.

Dr. Simon C. Watkins (University of Pittsburgh) will present "Novel
Probes and microscopies to dissect molecular processing in cystic fibrosis".

Dr. Nita Sahai ( University of Akron) will present

The Potential Role of Minerals in the Origins of Life

You can register on line at http://www.msneo.org/meetings.html

Hope to see you in the Fall meeting.

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Hello

I'd like to announce the next meeting of the Mountain States Society
of Electron Microscopists and Colorado Microbeam Analysis Society to
be held Thursday October 9 from 11A - 4P. This meeting breaks from
our traditional dinner meeting and allows for more talks and
networking time and has something for everyone.

In addition to the meeting there is an automated mineralogy by SEM/EDS
workshop held by Tescan at the Colorado School of Mines from 9A-11A.
Please contact Colleen Leary (cleary-at-tescan-usa.com or 724-772-7433)
for more details.

October 9, 2014 Table Mountain Inn, Golden CO

9:00-11:00A Automated Mineralogy Workshop by Jack Mershon,
Tescan. Held at Colorado School of Mines. Contact Colleen Leary
(cleary-at-tescan-usa.com or 724-772-7433) for details

11:00-11:15 Check In and Registration Table Mountain Inn

11:15-12:15 Lunch Buffet Chicken Tortilla Soup, ChimayÃ³ Chips
& Salsa, Dijon Mustard Potato Salad OR Tangy Fresh Made Coleslaw,
Assorted Wraps â€“ Roasted Turkey, Ham and Roast Beef â€“ Served with
Grilled Portobello Mushrooms, Hummus & Assorted Olives, Freshly Baked
Brownies & Cookies

12:30-1:15P Deborah Hall, Rush University Medical Center
â€œUnderstanding the Effects of Wear Particles: Lessons Learned from
Postmortem Retrievalsâ€

1:15-1:45P Brian Gorman, Colorado School of Mines
â€œDetermining the Properties of Oxides using Static and Dynamic Atom
Probe Tomographyâ€

1:45-2:00P Vendor Talks, Gatan, EDAX, JEOL

2:00-2:30 Coffee Break

2:30-3:15P Ed Vicenzi, Museum Conservation Institute,
Smithsonian Institution â€Examination of a 19th Century Daguerreotype
Photograph using High Resolution Scanning Transmission Electron
Microscopy for 2D and 3D Nanoscale Imaging and Analysisâ€

3:15-3:45P Jack Mershon, Tescan â€œHyperspectral Imaging of
Geological Thin Sectionsâ€

3:45-4:00P Adam Stokes, Colorado School of Mines â€œAtom Probe
Tomography Study on Cu(In,Ga)Se2 Grain Boundaries and Ordered Defect
Phase Transitionâ€

4:00P Adjourn

$25/professional $5/student

Please register online by October 3rd so we have an accurate number
for catering purposes. For online registration you can pay with a
credit card or choose to send a check payable to MSSEM/CoMAS, c/o John
Chandler, 2309 Cheyenne St, Golden CO 80401.

Registration link
http://microbeamanalysis.org/topical-conferences/colorado-mas-june-2014/registration-cmas-june-2014-meeting

Announcement link http://comas.geoloweb.ch/events.php

Thank you to our sponsors to date: Tescan, EDAX, Gatan, Jeol, Brukr,
Cameca, and Zeiss

--
~~~~~~~~~~~
Heather Lowers
Microbeam Lab
Lab: 303-236-3188
Off: 303-236-1184

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From dzexifto-at-iinet.net.au Tue Sep 30 08:45:57 2014
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Dear all

The Southern California Society for Microscopy and Microanalysis (SCSMM)
announce their Fall Meeting.

The meeting will be held on Thursday, October 23rd from 5:30 pm at the
Argyros Auditorium of Arnold and Mabel Beckman Center at the City of Hope,
Duarte, California. A buffet dinner will be served.

Our fall meeting will be dedicated to the Correlative Light and Electron
Microscopy. The featured speakers will be Thomas J. Deerinck (NCMIR and
UCSD), and Grayson Chadwick (Caltech).

Please see full program on our website: http://www.scsmm.org/

In order to register, Please RSVP by email at micromark-at-juno.com. Respond
no later than 5:00 p.m. Friday, October 17th.

The cost is $25 for professionals and $10 for students. Membership dues
can be paid at the door on October 23rd.

Nonmembers are welcome to attend.

Please pass along to those you feel would be interested. You can also find
us on the facebook

Look forward to seeing you at the meeting!

==============================Original Headers==============================
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12, 30 -- Subject: SCSMM fall meeting
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Hello

I have a student completing our EM program who has asked for some advice.

She has experience doing technical writing and will soon have a certificate in biological EM.

She asked if there were any career possibilities that could combine EM knowledge and technical writing skills.

What do I tell her?

Jon

Jonathan Krupp
Applied Science, Business & Technology
San Joaquin Delta College
5151 Pacific Ave.
Stockton, CA 95207
209-954-5284
jkrupp-at-deltacollege.edu

Find us on Facebook -at-
Electron Microscopy at SJ Delta College

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Email: caromill-at-iupui.edu
Name: Caroline A Miller

Organization: MSORV

Title-Subject: [Filtered] Fall Meeting

Message: Microscopy Society of the Ohio River Valley
Announces its 2014 Fall Meeting on October 23rd from 3-7:30 PM

This meeting will be held at Miami University, Oxford, OH, Center for
Advanced Microscopy and Imaging, Room 9 Upham Hall

Featuring Student Competition for Best Microscopy Presentation

Students present your work and have a chance to win a MSORV Travel Award
to the 2015 M&M meeting in Portland, OR for up to $1000.

Submission Deadline: Oct 6th, Send a 1 page abstract to Dave Tomlin,
dtomlin-at-azimuth-corp.com

For more information check out the MSORV website.

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Email: rashmi_mehata-at-yahoo.com
Name: Rashmi

Organization: DBT

Title-Subject: [Filtered] error message

Message: Hi!!

The FEI Quanta 200 showing the message "filament blown off" on the
contrary they are able to see the filament emission but the filament
emission signal turns from green to red.
The error occurs after some time when the HT and filament was switched
on. It is almost 3-4 four months now they are getting such erratic
problems.
Not able to trace out the reason for this.
Is it suitable to analyzed the samples in such conditions?

Regards
Rashmi

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Email: takenomm-at-uw.edu
Name: Marc Takeno

Organization: University of Washington

Title-Subject: [Filtered] [TEM] Experience (good/bad) with side-mounted
Olympus MegaviewG2 and Barlow CMOS100

Message: Hello,

We are considering purchasing a side-mounted TEM camera system for our
JEOL.

The candidates are:

Olympus Megaview G2
Barlow CMOS 100

If you have experiences good/bad with the above mentioned cameras,
including software and ease-of-use, please let me know! Thank you.

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From gaiihti-at-rr.com Wed Oct 1 17:29:10 2014
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Hi, Jon

Encourage her to investigate becoming a technical applications specialist. I started out my early days in this area and, over the intervening decades have found writing to be an invaluable skill to both my career development and my employers. I still write avidly (visit The Library at www.MicroscopyEducation.com) and have found that my writing always takes me into new and interesting applications and areas of technology.

Good hunting!
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Email: hwitkiewicz-at-ucsd.edu
Name: Halina Witkiewicz

Organization: PRISM

Title-Subject: [Filtered] Re: [TEM] Experience (good/bad) with
side-mounted Olympus MegaviewG2 and Barlow CMOS100

Message: Hi Marc,

My experience with side-mounted Olympus Megaview III camera was GOOD.
Please see http://f1000research.com/search?q=witkiewicz ('Viewing'
section in any of the three articles listed there).

Regards,
Halina

Email: takenomm-at-uw.edu
Name: Marc Takeno

Organization: University of Washington

Title-Subject: [Filtered] [TEM] Experience (good/bad) with side-mounted
Olympus MegaviewG2 and Barlow CMOS100

Message: Hello,

We are considering purchasing a side-mounted TEM camera system for our
JEOL.

The candidates are:

Olympus Megaview G2
Barlow CMOS 100

If you have experiences good/bad with the above mentioned cameras,
including software and ease-of-use, please let me know! Thank you.

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What kind of conditions are needed to deposit a film of thickness of about 10 nm by evaporation of a tungsten wire in high vacuum by resistive heating? What level of vacuum is necessary to avoid oxidation, what kind of power supply, W wire thickness etc. parameters are needed? What would be the approximate rate of deposition?

Krassimir Bozhilov.

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Hi

I would guess that there are a number of reasons why you have this problem,
others may help here? However, provided you have stable emission and the
focus is also stable will have no problems with your work.

Steve

Steve Chapman FRMS
Protrain for Consultancy and Courses in Electron Microscopy
Mobile +44 (0) 7711 606967 Fax +44 (0)1280 814007
www.emcourses.com

X-from: rashmi_mehata-at-yahoo.com ()

Email: rashmi_mehata-at-yahoo.com
Name: Rashmi

Organization: DBT

Title-Subject: [Filtered] error message

Message: Hi!!

The FEI Quanta 200 showing the message "filament blown off" on the contrary
they are able to see the filament emission but the filament emission signal
turns from green to red.
The error occurs after some time when the HT and filament was switched on.
It is almost 3-4 four months now they are getting such erratic problems.
Not able to trace out the reason for this.
Is it suitable to analyzed the samples in such conditions?

Regards
Rashmi

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23, 22 -- From protrain-at-emcourses.com Thu Oct 2 21:25:17 2014
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From kjooiiio-at-memoryplay.com Fri Oct 3 04:22:19 2014
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Message-ID: {BBAB8827.8945693-at-memoryplay.com}

I did not have a change to dig into Quanta 200, but based on typical
architecture of SEM detection of blown filament should be made by the
high voltage power supply (HVPS) - usually a pricey item to repair.

Can't be sure of exact multifunction, but if parameters of filament
current, extractor/anode currents, and the image are stable, then I'd
run it until the HVPS quits completely.

Valery Ray - also with NISP, UMDCP Nanocenter
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UMD E-Mail: vray-at-umd.edu
Web: www.partbeamsystech.com
Web: www.freudlabs.com

On 10/2/2014 10:26 PM, protrain-at-emcourses.com wrote:
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} Hi
}
} I would guess that there are a number of reasons why you have this problem,
} others may help here? However, provided you have stable emission and the
} focus is also stable will have no problems with your work.
}
} Steve
}
} Steve Chapman FRMS
} Protrain for Consultancy and Courses in Electron Microscopy
} Mobile +44 (0) 7711 606967 Fax +44 (0)1280 814007
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} X-from: rashmi_mehata-at-yahoo.com ()
}
} Email: rashmi_mehata-at-yahoo.com
} Name: Rashmi
}
} Organization: DBT
}
} Title-Subject: [Filtered] error message
}
} Message: Hi!!
}
} The FEI Quanta 200 showing the message "filament blown off" on the contrary
} they are able to see the filament emission but the filament emission signal
} turns from green to red.
} The error occurs after some time when the HT and filament was switched on.
} It is almost 3-4 four months now they are getting such erratic problems.
} Not able to trace out the reason for this.
} Is it suitable to analyzed the samples in such conditions?
}
} Regards
} Rashmi
}
}
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} 23, 22 -- From protrain-at-emcourses.com Thu Oct 2 21:25:17 2014
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4, 45 -- CC: microscopy-at-microscopy.com
4, 45 -- Subject: Re: [Microscopy] FW: viaWWW: Filament error message
4, 45 -- References: {201410030226.s932Quft028088-at-ns.microscopy.com}
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Contents Retrieved from Microscopy Listserver Archives
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It appears our HT tank on our CM12 needs replacing. Has anyone ever had to
replace or self service by opening up the tank and checking/replacing bad
capacitors? Any suggestions specific for the CM12 would be appreciated

Fred Hayes
Manager, AMCaT Labs
Univ of CA Davis
Dept Chem Engr and Mat Sci
3001 Ghausi Hall
1 Shields Ave
Davis, CA 95616-5270
http://chms.engineering.ucdavis.edu/research/amcat/

==============================Original Headers==============================
4, 24 -- From fahayes-at-ucdavis.edu Fri Oct 3 13:41:27 2014
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4, 24 -- From: "Fred Hayes" {fahayes-at-ucdavis.edu}
4, 24 -- To: {Microscopy-at-microscopy.com}
4, 24 -- Subject: CM12 TEM HT tank troubleshooting
4, 24 -- Date: Fri, 3 Oct 2014 11:41:24 -0700
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Fred, please call, we can help you.

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

On 10/3/2014 2:42 PM, fahayes-at-ucdavis.edu wrote:
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}
} It appears our HT tank on our CM12 needs replacing. Has anyone ever had to
} replace or self service by opening up the tank and checking/replacing bad
} capacitors? Any suggestions specific for the CM12 would be appreciated
}
} Fred Hayes
} Manager, AMCaT Labs
} Univ of CA Davis
} Dept Chem Engr and Mat Sci
} 3001 Ghausi Hall
} 1 Shields Ave
} Davis, CA 95616-5270
} http://chms.engineering.ucdavis.edu/research/amcat/
}
}
}
} ==============================Original Headers==============================
} 4, 24 -- From fahayes-at-ucdavis.edu Fri Oct 3 13:41:27 2014
} 4, 24 -- Received: from msa2.ucdavis.edu (msa2.ucdavis.edu [128.120.32.182])
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} 4, 24 -- From: "Fred Hayes" {fahayes-at-ucdavis.edu}
} 4, 24 -- To: {Microscopy-at-microscopy.com}
} 4, 24 -- Subject: CM12 TEM HT tank troubleshooting
} 4, 24 -- Date: Fri, 3 Oct 2014 11:41:24 -0700
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4, 27 -- From vitalylazar-at-att.net Fri Oct 3 15:30:10 2014
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4, 27 -- To: fahayes-at-ucdavis.edu, microscopy Listserver {Microscopy-at-microscopy.com}
4, 27 -- Subject: Re: [Microscopy] CM12 TEM HT tank troubleshooting
4, 27 -- References: {201410031842.s93Igs0I020025-at-ns.microscopy.com}
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Dear listers,

Electropolishing refractory metals, particularly tungsten, has a reputation as non-trivial. The literature consensus is a weak NaOH solution, but consensus degenerates after that.

Does anyone have advice for techniques and/or hardware? My interest is both bulk SEM electropolishing and 3 mm twin-jet for TEM.

Vendor responses welcome.

Chad

---------------------
Chad M. Parish, Ph.D.
Research and Development Staff Member
Materials Science and Technology Division
Oak Ridge National Laboratory
Phone: 1 865 574 0092
Email: parishcm-at-ornl.gov

==============================Original Headers==============================
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Email: Richard.Gursky-at-stjude.org
Name: Richard Gursky

Organization: St. Jude ChildrenÂ’s Research Hospital

Title-Subject: [Filtered] User Friendly Laboratory Information
Management System (LIMS)

Message: User Friendly Laboratory Information Management System.

We are looking to replace our curret Laboratory Information Management
System (LIMS) for a more User Friendly LIMS (read as takes less time for
Sample Logging & Cost Center Charging). Would any of you recommend the
system that you work with? Could someone give me any information on the
LIMS that you use and could you put me in touch with your IT guys so we
could get some information from them?

Thanks Much!

Richard Gursky
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Email: j.knowles-at-ucl.ac.uk
Name: Jonathan

Organization: UCL

Title-Subject: [Filtered] TEM camera software

Message: Hi

We have a Hamamatsu camera and were running the AMT imaging software for
image capture. Both pretty old verions on Xp!

The PC has died and we needed to reinstall the software, but AMT are
quoting a hefty price to come in and install it.

Would anyone by any chance know where i could get a copy at a reasonable
price or free?

thanks

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Email: trrajaprakash1-at-u.northwestern.edu
Name: Rajaprakash Ramachandramoorthy

Organization: Northwestern University

Title-Subject: [Filtered] TEM phosphor screen

Message: TEM Specification: JEOL 2100F

Question: What is the approximate response time of the phosphor screen
(viewing green screen), which I believe is commonly coated with Zinc
Sulphide? I need to know the rise and decay time of ZnS fluorescence. Is
there a way for me to characterize this response time?

Also, can this response time be varied with different coatings or by
varying the intensity of the beam?

Thank you in advance,

Raj.

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From reykarpu-at-jajagroup.com Sat Oct 4 09:45:45 2014
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Message-ID: {F21C2DD9.3552688-at-jajagroup.com}

Greetings Microscopists!

Recently I have been informed that Leica will no longer be carrying the 200 micron membrane carriers for their EM-Pact(1 or 2) High Pressure Freezer. I am wondering if anyone has some laying around that they would be willing to sell or give away because they are no longer using? Or maybe they have something better that they use as a replacement.

Has anyone else encountered this problem, and do they have a solution?

Cheers!

Rhonda Trimble Ross
Electron Microscopy Specialist
Stowers Institute
1000 E 50th St
Kansas City, MO 64110
rra-at-stowers.org

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Email: meb187-at-pitt.edu
Name: Margaret Bisher

Organization: University of Pittsburgh School of Medicine

Title-Subject: [Filtered] Forma Scientific Incubator, Model 3546

Message: We have inherited this incubator and I was wondering if anyone might have the manual.

Thanks for checking.

Margaret E. Bisher
Department of Cell Biology
University of Pittsburgh School of Medicine
346 South Biomedical Science Tower
Pittsburgh, PA 15261

Lab: (412) 648-9565
Email: meb187-at-pitt.edu

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From gaiweji-at-tmodns.net Thu Oct 9 01:29:04 2014
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Just curious. How essential is it to prepare CO2 free water for mixing and using lead citrate stain?

I have always done it, but wonder if anyone has tried skipping it.

How about alternate ways to prepare it. Boiling is the standard, but anyone boil using microwave or I have heard sonicating might work too.

Sitting here with a scalding flask of water, wondering if I can cheat the EM gods.

Jon

Jonathan Krupp
Applied Science, Business & Technology
San Joaquin Delta College
5151 Pacific Ave.
Stockton, CA 95207
209-954-5284
jkrupp-at-deltacollege.edu

Find us on Facebook -at-
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Jon - What is the downside of doing this just to be sure? There aren't many steps in science easier than "boil water". I have used a microwave but to ensure long enough boiling, it makes it a little messier since the water spills over so a hot plate is probably worth the effort. I am sure sonication works but that is more trouble than needed since you don't have to worry about evaporative loss or flammability when you boil water (unless your lab is adjacent to a fracking well). I have used sonication in the steps involving getting lead nitrate or lead citrate to go into solution but only since I am so compulsive. Could you cheat the gods and skip this step? Sure, 99 times out of a 100. But the downstream problems aren't worth the risk. If you made a batch and got staining artefacts, how would you know if you screwed up the staining step or the error was in not originally boiling the water. I am as frugal and lazy as the next guy but this is as cheap and easy as it gets in science. Find something else to eliminate in your life. Tom

Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
phillipst-at-missouri.edu

http://biology.missouri.edu/people/?person=88
http://www.biotech.missouri.edu/mcc/

-----Original Message-----
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To: Phillips, Thomas E.

Just curious. How essential is it to prepare CO2 free water for mixing and using lead citrate stain?

I have always done it, but wonder if anyone has tried skipping it.

How about alternate ways to prepare it. Boiling is the standard, but anyone boil using microwave or I have heard sonicating might work too.

Sitting here with a scalding flask of water, wondering if I can cheat the EM gods.

Jon

Jonathan Krupp
Applied Science, Business & Technology
San Joaquin Delta College
5151 Pacific Ave.
Stockton, CA 95207
209-954-5284
jkrupp-at-deltacollege.edu

Find us on Facebook -at-
Electron Microscopy at SJ Delta College

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I got an old Hitachi s-450 SEM with EDX from a friend when he upgraded to a Jeol 6300F.

It has some scan problems which I dont think will be much of an issue, the problem is I do not have any good filaments. The last good one he had blew not long ago and I am a bit reluctant to blow $500 on a box of filaments if it turns out to be a bigger problem than I think it is. Anyone know where I might find a seller that sells in individual quantities.

I have a couple bases to rebuild but I would like to find a couple more, I have a capacitive discharge spot welder to rebuild my own once I get some tungsten wire.

Thanks

-Jerry

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Dear Microscopy Managers and Directors
Please share with your light and fluorescence microscopy core users

Thanks to a grant from Olympus Corporation there are tuition
scholarships available for US based graduate students and post-docs for the
Advanced Microscopy LIVE Workshop

January 12-16, 2015
University of California, Santa Barbara

Offered by the Neuroscience Research Institute (NRI) and Department of
Molecular, Cellular, and Developmental Biology (MCDB).
contact Mary Raven m_raven-at-lifesci.ucsb.edu for scholarship information
and requests

See for workshop information:
http://microscopy.nri.ucsb.edu/education/imaging-live-workshop/advanced-microscopy-live

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Jon:

Agreeing with Philip, but with two additionals. I have had a couple students
mix it up without CO2 Freewater only to have them then come and ask me "So is
this Lead percipitate?" Yup, seems silly but in these cases they wind up with very
rapid percipiation, which I assume was lead carbonate. (If they waited long enough
and were careful they could "stain" with the lead free solution and avoid the
percipitate . . . but since the lead seems wind up on the bottom and sides of the
containers why?)

You can also make CO2 free water with an autoclave. But since most
autoclaves either reek or are heated with steam from decades old plumbing lines
and reek, I 've never trusted the results to be clean enough to try staining with.

Sounds like an M&M poster to me!

I just overfill the flask so it won't run dry, and I have plenty for the Pb-Cit and
the NaOH wash water. Set it to boil, putter around cleaning optics, dishes, dusting,
putting away glassware, sorting old samples and doing the 50,000 other little lab
tasks until its throughly boiled.

On 10 Oct 2014 at 15:13, jkrupp-at-deltacollege.edu wrote:

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} Just curious. How essential is it to prepare CO2 free water for mixing
} and using lead citrate stain?
}
} I have always done it, but wonder if anyone has tried skipping it.
}
} How about alternate ways to prepare it. Boiling is the standard, but
} anyone boil using microwave or I have heard sonicating might work too.
}
} Sitting here with a scalding flask of water, wondering if I can cheat
} the EM gods.
}
} Jon
}
} Jonathan Krupp
} Applied Science, Business & Technology
} San Joaquin Delta College
} 5151 Pacific Ave.
} Stockton, CA 95207
} 209-954-5284
} jkrupp-at-deltacollege.edu
}
} Find us on Facebook -at-
} Electron Microscopy at SJ Delta College
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} Headers============================== 16, 38 -- From
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I could not agree more, Jon.

And would add: http://www.ou.edu/research/electron/bmz5364/sato.pdf that lasted without mishap for more than the estimated year at room temp, in the dark in 20ml brown bottles with a PTFE lined screw cap. BUT, in spite of the chemistry, I still boiled the water.

Cheers to all who are also weighed down by the vagaries of toxic lead that causes huge frustrated sighs of toxic gas.

Fred

Frederick C. Monson, PhD
Technical Director
Center for Microanalysis and Imaging, Research and Training (CMIRT)
West Chester University of Pennsylvania
Schmucker Science South - Room SSS-024
MailDrop: Geology-Astronomy
750 South Church Street
West Chester, PA, 19383
610-738-0437
fmonson-at-wcupa.edu

-----Original Message-----
X-from: PhillipsT-at-missouri.edu [mailto:PhillipsT-at-missouri.edu]
Sent: Friday, October 10, 2014 5:27 PM
To: Monson, Frederick

Jon - What is the downside of doing this just to be sure? There aren't many steps in science easier than "boil water". I have used a microwave but to ensure long enough boiling, it makes it a little messier since the water spills over so a hot plate is probably worth the effort. I am sure sonication works but that is more trouble than needed since you don't have to worry about evaporative loss or flammability when you boil water (unless your lab is adjacent to a fracking well). I have used sonication in the steps involving getting lead nitrate or lead citrate to go into solution but only since I am so compulsive. Could you cheat the gods and skip this step? Sure, 99 times out of a 100. But the downstream problems aren't worth the risk. If you made a batch and got staining artefacts, how would you know if you screwed up the staining step or the error was in not originally boiling the water. I am as frugal and lazy as the next guy but this is as cheap and easy as it gets in sc!

ience. Find something else to eliminate in your life. Tom

Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
phillipst-at-missouri.edu

http://biology.missouri.edu/people/?person=88
http://www.biotech.missouri.edu/mcc/

-----Original Message-----
X-from: jkrupp-at-deltacollege.edu [mailto:jkrupp-at-deltacollege.edu]
Sent: Friday, October 10, 2014 2:47 PM
To: Phillips, Thomas E.

Just curious. How essential is it to prepare CO2 free water for mixing and using lead citrate stain?

I have always done it, but wonder if anyone has tried skipping it.

How about alternate ways to prepare it. Boiling is the standard, but anyone boil using microwave or I have heard sonicating might work too.

Sitting here with a scalding flask of water, wondering if I can cheat the EM gods.

Jon

Jonathan Krupp
Applied Science, Business & Technology
San Joaquin Delta College
5151 Pacific Ave.
Stockton, CA 95207
209-954-5284
jkrupp-at-deltacollege.edu

Find us on Facebook -at-
Electron Microscopy at SJ Delta College

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Dear Listserver,
We have a strange phenomenon occurring where RBC are appearing both light
and dark in our TEM sections. This occurs with or without staining,
In both light and electron microscopy preps. The cells have been plasmodium
infected, and processed as follows:
2.5% Glutaraldehyde fixed at least overnight (against our wishes as I
recommend a Paraform. Glutaraldehyde mixture for any infected material)
1% Osmicated followed by graded ethanol dehydrations and Epon embedding.
Sections are 80 nm thick and normally stained with UA followed by Pb.
Any suggestions or references would be highly appreciated.

Sincerely,
Michael Delannoy

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4, 20 -- Subject: light vs dark RBC
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Good morning (all),
Michael,
perhaps only talking at large....
this phenomenon perhaps (quite) "normal" due to physiological condition of the RBC'S in a state before, during and after infection with plasmodium(?)
"Dark" and "light" morphology of RBC's has been shown by micrographs (description ??) in/by:

Hannes Wickert, Georg Krohne
Trends in Parasitology, Volume 23, Issue 10, October 2007, Pages 502-509
The complex morphology of Maurer's clefts: from discovery to three-dimensional reconstructions
see: http://www.sciencedirect.com/science/article/pii/S1471492207002218

see:
Ultrastructure of the lung in a murine model of malaria-associated acute lung injury/acute respiratory distress syndrome
Aitken et al. Malaria Journal 2014, 13:230 Page 5 of 10, Fig.4
http://www.malariajournal.com/content/13/1/230
http://www.malariajournal.com/content/pdf/1475-2875-13-230.pdf

This out of my collected files,
it might be there are other reports on ultrastructural morphology on severe search for this phenomenon.
Best wishes, good luck,
Wolfgang

(Wolfgang MUSS, SALZBURG, AUSTRIA)

} -----Ursprüngliche Nachricht-----
} Von: mdelann1-at-jhmi.edu [mailto:mdelann1-at-jhmi.edu]
} Gesendet: Dienstag, 14. Oktober 2014 21:25
} An: Muß Wolfgang
} Betreff: [Microscopy] Light vs dark RBC [in TEM, plasmodium infected]
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} Dear Listserver,
}
} We have a strange phenomenon occurring where RBC are appearing both
} light and dark in our TEM sections.
} This occurs with or without staining, In both light and electron
} microscopy preps.
} The cells have been plasmodium infected, and processed as follows:
} 2.5% Glutaraldehyde fixed at least overnight (against our wishes as I
} recommend a Paraform. Glutaraldehyde mixture for any infected material)
} 1% Osmicated followed by graded ethanol dehydrations and Epon
} embedding.
} Sections are 80 nm thick and normally stained with UA followed by Pb.
} Any suggestions or references would be highly appreciated.
}
} Sincerely,
} Michael Delannoy
}
}
}
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Hi Everyone,

We have a new postdoctoral position opening up for an image analyst
dedicated to Light sheet microscopy. Please see

https://www.recruit.ox.ac.uk/pls/hrisliverecruit/erq_jobspec_version_4.display_form

Postdoctoral Image Analyst in Light Sheet Microscopy
Department of Biochemistry, South Parks Road, Oxford
Grade 8: Â£38,511 - Â£45,954 p.a.

We are seeking to appoint a Postdoctoral Image Analyst in Light Sheet
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Contact Person : Margaret Dixon or Rita Emberton Vacancy ID : 115368
Contact Phone : 01865 613204 Closing Date : 10-Nov-2014 Contact Email :
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Thanks,

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--
Ian Dobbie
Micron Bioimaging Unit Manager,
Biochemistry,
University of Oxford,
South Parks Road,
Oxford
OX1 3QU
Tel: 01865 613323
Email: ian.dobbie-at-bioch.ox.ac.uk

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Hello Everyone,
Size does matter, at least for fonts. I'm running Gatan TEM camera model 780 duel View on my TEM. It's a great improvement from the previous camera, but I would like to increase the font size of the global session information. I can click on each field and change the font size, but doing each of 7 field for each image is too time consuming.

I'm hope one of the Gatan 780 user can advise me on how to globally increase font size.

Thanks!

Frank

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==============================Original Headers==============================
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Message: Job Description: CRYO SAMPLE AND SEM SPECIALIST, Koch Institute for Integrative Cancer
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to develop cryo preservation techniques for a wide variety of novel samples. Will bring creativity
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Job Requirements: REQUIRED: a bachelorÂ’s degree (masterÂ’s or Ph.D. preferred); at least six years of
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Title-Subject: [Filtered] ElectroScan 2020

Message: We are decommissioning an ElectroScan 2020 SEM. It has no CPU, no vacuum unit and many
boards have been taken. Does anyone have any need for any other parts from it? You pay for
shipping and you can have what you need. We are located in Michigan. I have unused 20 um, 30 um,
and 50 um apertures. I have a good LaB6 filament. Unrelated but also free I have Tungsten wire
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Hi Frank,

You may wish to check out some of the DM scripting websites such as Dave
Mitchell's site (http://www.dmscripting.com) or the Univ of Graz site
(http://portal.tugraz.at/portal/page/portal/felmi/DM-Script). Dave has
a script for adding an adjustable scale bar that you may be able to
modify for your purpose.
(http://www.dmscripting.com/add_adjustable_scale_bar.html) He als has a
"scale bar control" which may be useful.
(http://www.dmscripting.com/scalebarcontrol.html).

Good luck and let us know if you get a working script!

Cheers,
Henk

On 10/15/2014 9:59 AM, frank_karl-at-ardl.com wrote:
}
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} Hello Everyone,
} Size does matter, at least for fonts. I'm running Gatan TEM camera model 780 duel View on my TEM. It's a great improvement from the previous camera, but I would like to increase the font size of the global session information. I can click on each field and change the font size, but doing each of 7 field for each image is too time consuming.
}
} I'm hope one of the Gatan 780 user can advise me on how to globally increase font size.
}
} Thanks!
}
} Frank
}
}
}
}
} This email and any of its attachments may contain confidential information intended only for the use of the addressee(s). If the reader of this email is not the intended recipient or the employee or agent responsible for delivering it to the intended recipient, you are hereby notified that any dissemination or copying of this email is strictly prohibited. If you have received this email in error, please notify us by return email at info-at-ardl.com, permanently delete the email, and destroy any printouts. If this email contains test data and/or draft reports, you are hereby notified that only a signed original test report will contain official results, a copy of which resides in the project folder located at ARDL, Inc. Thank you. Akron Rubber Development Laboratory, Inc.
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} 10, 28 -- From frank_karl-at-ardl.com Wed Oct 15 08:57:56 2014
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--

Hendrik O. Colijn

*C*enter for *E*lectron *M*icroscopy & *A*nalysi*S*

1305 Kinnear Rd, Suite 100, Columbus, OH 43212

colijn.1-at-osu.edu {mailto:colijn.1-at-osu.edu} 614.643.3458 Office

cemas.osu.edu

"Time is that quality of nature which keeps things from happening all at
once. Lately it doesn't seem to be working."

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***************************************************************************************
Forwarded from "Ask a Microscopist"
Please remember that the person asking the question is likely
not a member the listserver, and
**any reply should go directly to the poster**
as well as to the list.
Using the "reply" function in your email does *not* send your answer
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} Below is the result of your form, submitted on Thursday, October 16, 2014 at 12:13:47 AM.
}
} realname - Zhongyuan Zuo
} Email - zhongyuanzuo-at-hotmail.com
} ORGANIZATION - Baylor College of Medicine
} EDUCATION - Graduate College
} LOCATION - Houston
} SUBJECT_OF_QUESTION - Ultramicrotome
} QUESTION - Hi,
} I just join the lab in Baylor as an EM specialist. I am having difficulties with the concentric region finding for Drosophila. I need both Retina and Lamina region from the very centered region in order to get a good section. Any suggestions on alignment of the UC 7 microtome? Also, I have in consistency of the thickness in both inter and intera sections. Please help me. Thank you!

==============================Original Headers==============================
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Dear All,

We are disposing some equipment that is used with NanoScope II PC's:

CRT VGA monitor for image display

CRT Monochrome monitor for Control display

Interface boards for monochrome monitors

miscellaneous boards for controllers

We are disposing some general purpose video equipment:

-CRT monitors that display NTSC composite video (typically from an optical
microscope).

All of this equipment is available for free pickup at our lab in
Indianapolis. We need to move it out to make room for more valuable
equipment.

regards,
Don
=============================================
Don Chernoff, Ph.D., President
Advanced Surface Microscopy, Inc.; 3250 N. Post Rd., Suite 120;
Indianapolis IN 46226 USA
E-Mail: donc-at-asmicro.com www.asmicro.com
Voice: 317-895-5630 Toll free: 800-374-8557 (in USA & Canada)
Fax: 317-895-5652
[business activities since 1990: analytical services in AFM, AFM probes,
consulting, training, calibration and test specimens,
calibration and measurement software, used NanoScope equipment.]
=============================================

==============================Original Headers==============================
15, 36 -- From donc-at-asmicro.com Thu Oct 16 12:40:48 2014
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15, 36 -- From: "Don Chernoff at ASM" {donc-at-asmicro.com}
15, 36 -- To: "Microscopy List" {microscopy-at-microscopy.com}
15, 36 -- Subject: Free NanoScope II and general purpose video equipment
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Dear Justin,

We don't have the NanoScope II controller, but we do have STM and AFM
equipment that is compatible with NanoScope II. We also sell used or
refurbished later model NanoScope controllers, complete systems, etc.
regards,
Don
=============================================
Don Chernoff, Ph.D., President
Advanced Surface Microscopy, Inc.; 3250 N. Post Rd., Suite 120;
Indianapolis IN 46226 USA
E-Mail: donc-at-asmicro.com www.asmicro.com
Voice: 317-895-5630 Toll free: 800-374-8557 (in USA & Canada)
Fax: 317-895-5652
[business activities since 1990: analytical services in AFM, AFM probes,
consulting, training, calibration and test specimens,
calibration and measurement software, used NanoScope equipment.]
=============================================
----- Original Message -----
From: Justin A. Kraft
To: donc-at-asmicro.com
Sent: Thursday, October 16, 2014 1:56 PM
Subject: [a] Re: [Microscopy] Free NanoScope II and general purpose video
equipment

Is the nanoscope available?

} On Oct 16, 2014, at 8:51 PM, donc-at-asmicro.com wrote:
}
}
}
}
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of
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} Dear All,
}
}
}
} We are disposing some equipment that is used with NanoScope II PC's:
}
} CRT VGA monitor for image display
}
} CRT Monochrome monitor for Control display
}
} Interface boards for monochrome monitors
}
} miscellaneous boards for controllers
}
} We are disposing some general purpose video equipment:
}
} -CRT monitors that display NTSC composite video (typically from an
optical
} microscope).
}
} All of this equipment is available for free pickup at our lab in
} Indianapolis. We need to move it out to make room for more valuable
} equipment.
}
}
}
} regards,
} Don
} =============================================
} Don Chernoff, Ph.D., President
} Advanced Surface Microscopy, Inc.; 3250 N. Post Rd., Suite 120;
} Indianapolis IN 46226 USA
} E-Mail: donc-at-asmicro.com www.asmicro.com
} Voice: 317-895-5630 Toll free: 800-374-8557 (in USA & Canada)
} Fax: 317-895-5652
} [business activities since 1990: analytical services in AFM, AFM
probes,
} consulting, training, calibration and test specimens,
} calibration and measurement software, used NanoScope equipment.]
} =============================================
}
}
} ==============================Original
Headers==============================
} 15, 36 -- From donc-at-asmicro.com Thu Oct 16 12:40:48 2014
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} 15, 36 -- Message-ID: {9CE2A34655B24105B08F4E31C3FD8CF0-at-asm20}
} 15, 36 -- From: "Don Chernoff at ASM" {donc-at-asmicro.com}
} 15, 36 -- To: "Microscopy List" {microscopy-at-microscopy.com}
} 15, 36 -- Subject: Free NanoScope II and general purpose video equipment
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Hi Zhongyuan,

I have not done any of this recently, and is therefore not fresh in my mind, but I remember that good fixation, embedding and therefore sectioning of arthropod eyes can be challenging. Samples with highly variable internal density always seem to be tricky. The exoskeleton can form an excellent barrier to ingress of chemicals (at least in the much larger spiders that I dealt with). I suspect the key will be in the sample preparation rather than the settings on the microtome.

For sample prep, partial dissection, punching little holes etc. all helps. I have talked to people (admittedly dealing with even larger more armoured arthropods) that use DMSO in some of their solutions.

In order to get the best help from people on this forum, you may have to describe the problem you are having in more detail and keep in mind that not everyone, including I, will understand what "the concentric region" is exactly.

Kind regards
Duane

AgResearch Limited | Lincoln Research Centre | New Zealand
Dr Duane P Harland, Senior Scientist
T +64 3 321 8710 E duane.harland-at-agresearch.co.nz

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} Below is the result of your form, submitted on Thursday, October 16, 2014 at 12:13:47 AM.
}
} realname - Zhongyuan Zuo
} Email - zhongyuanzuo-at-hotmail.com
} ORGANIZATION - Baylor College of Medicine EDUCATION - Graduate College
} LOCATION - Houston SUBJECT_OF_QUESTION - Ultramicrotome QUESTION - Hi,
} I just join the lab in Baylor as an EM specialist. I am having difficulties with the concentric region finding for Drosophila. I need both Retina and Lamina region from the very centered region in order to get a good section. Any suggestions on alignment of the UC 7 microtome? Also, I have in consistency of the thickness in both inter and intera sections. Please help me. Thank you!

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Title-Subject: [Filtered] Digital counter for filament usage

Message: Hi All,
Is there any time counter for filament on/off available with FEI or third party?

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Title-Subject: [Filtered] Webinar: Optimizing STEM Spectrum Image Acquisition for High-Speed Analysis

Message: If you are a EELS or EFTEM user, you may be interested in attending this 30 minute webinar.
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Ravi,

I assume your FEI scope has a computer interface - I keep track of
filament use with a Macro Express macro hooked to a command icon (also
through macro express) for our JEOL 5600. When I click the icon, the
macro starts/stops the microscope high tension and writes a timestamp to
a text file, calculating elapsed session time and total beam time when
the HT is turned off. Fairly simple, really. I can send you details if
you're interested.

Macro Express isn't free, but well worth the trivial amount it costs. It
automates all kinds of things I do routinely on scope computers and
elsewhere. See http://www.macros.com/

No commercial interest, just a happy customer.

Cheers,

Jim

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Digital Microscopy Facility
Mount Allison University
63B York St.
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CANADA

phone: 506-364-2519
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email: jehrman-at-mta.ca
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For Sale: Parachute. Only used once,
never opened, small stain.

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Hi Ravi,

When the HT is turned on, the 12 volt filament supply is powered up. If
you take a 12 volt timer and tie it to the power supply, it will give
you the HT ON time. Since we turn the HT off between users, it served
us well as a usage timer.

Henk

On 10/17/2014 9:10 AM, microscopylistserver-noreply-at-microscopy.com wrote:
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--

Hendrik O. Colijn

*C*enter for *E*lectron *M*icroscopy & *A*nalysi*S*

1305 Kinnear Rd, Suite 100, Columbus, OH 43212

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"Time is that quality of nature which keeps things from happening all at
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Why do you ask?
Do you have a W gun or a field emission gun?

If you are seeking to track filament lifetime, that can probably be done rather easily if you can find the right connection to a circuit showing when the high voltage is on. I don't know why you would want to do that with a field emission gun.

If you are seeking to track hours for billing I can understand that and there are multiple ways to do it.
- You might be able to tie into a circuit that show voltage when the high voltage is on. There are hour meters available through McMaster Carr and other outlets than run when a voltage is applied to them.
- We have considered having a program run in front of the user interface so that users need to log in to clear it and access the UI. That could give us a lot more information.
- We have also heard of someone putting a box inline with the monitor power. The monitor can only be energized when someone logs into another monitor computer.

Warren
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Title-Subject: [Filtered] Digital counter for filament usage

Message: Hi All,
Is there any time counter for filament on/off available with FEI or third party?

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All,

Online registration and abstract submission are now open for AMAS XIII, the 13th biennial Australian Microbeam Analysis Symposium, which will be held in Hobart, Tasmania, on 11-13 February 2015, with Pre-Meeting Workshops 9-10 Feb, 2015.

The AMAS symposia provide a forum to discuss and share ideas on advances, trends and challenges in microanalysis and imaging with national and international leaders in the field, with an emphasis on practical solutions and applications.

Please see our website

http://www.microscopy.org.au/amas/amas13/

for more information such as second circular, invited speakers, workshop program, registration,
accommodation, and student travel bursary information.

Please feel free to forward this email to your colleagues and anyone that you think might be
interested in attending.

Many thanks,

Karsten

AMAS XIII Co-Chair

Dr Karsten Goemann
Research Fellow, Electron Microscopy & X-Ray Microanalysis
Central Science Laboratory, University of Tasmania
Mail: Private Bag 74, Hobart TAS 7001, Australia
Location: Rooms 254-256, Chemistry Building, Dobson Road, Sandy Bay TAS 7005, Australia
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Hi,

Recently one of our users asked about how to prepare synthetic scaffolds with cells attached. The material make the scaffolds dissolves in ethanol. So is there any way to avoid using ethanol but using something else (e.g. Acetone) and proceed with critical point drying? Or we might have to do freeze drying?

Any advice will be appreciated!

Xuanhao Sun, Ph.D.

Bioscience Electron Microscopy Laboratory
University of Connecticut
BPB G06, Unit 3242
Storrs, CT 06269-3242

Office Tel: 860-486-6368
Lab Tel: 860-486-2914
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Recently one of our users asked about how to prepare synthetic scaffolds
with cells attached. The material used to make the scaffolds dissolves in
ethanol. So is there any way to avoid using ethanol but using something
else (e.g. Acetone) and proceed with critical point drying? Or we might
have to do freeze drying?

Any advice will be appreciated!

Xuanhao Sun, Ph.D.
Bioscience Electron Microscopy Laboratory
University of Connecticut
BPB G06, Unit 3242
Storrs, CT 06269-3242
Office Tel: 860-486-6368
Lab Tel: 860-486-2914
Fax: 860-486-6369
website: emlab.uconn.edu
=======================

Dear Xuanhao,

Yes, you may critical point dry in Acetone but before you do I'd ask the
investigators if they have tried acetone on the scaffolds for it may
dissolve them too. Another question would be have they fixed the material
with glutaraldehyde followed by osmium tetroxide before putting it into
ethanol? If it becomes stable it could eliminate your problem.

The second point is that the vapors from the CPD need to be vented into a
chemical hood so that you are not exposed to the acetone which one should
not breathe. Run off a copy of the MSDS sheet for safety recommendations.

The third point is that you will need "dry acetone". Do not use acetone
from a big old jug! I have successfully used Mallinckrodt 2440 Analytical
reagent grade. It has a water content of 0.2%. It is available in pint
bottles that are good for they do not set around for a long time before
being used up like the gallons do. I had made the dilutions from it and of
course used it in the CPD chamber at 100%. I believe that several of the
EM suppliers also supply acetone that is "dry".

I have heard of using amyl acetate in the CPD but know that the old Sorval
CPD instructions said that the O-rings needed to be changed to a different
material for the standard ones would not hold up. Amyl acetate is also
nasty and must be vented to a hood. I do not recall what was used for the
dehydration before putting the samples into amyl acetate for I had not
used it.

HMDS is sometimes used in place of CPD but ethanol is used for dehydration
up to 100% so that is not a good substitute.

Opinions and experiences related are those of Pat Connelly and do not
represent the NIH.
This message is not confidential and can be freely shared and reproduced.

Pat

Patricia Stranen Connelly
Research Assistant
NHLBI EM Core Facility
National Institutes of Health
Building 14E Room 111B
Bethesda, MD 20892-5570
301-496-3491
connellyps-at-mail.nih.gov

http://www.nhlbi.nih.gov/research/intramural/researchers/core/electron-micr
oscopy-core/index.html

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I had a question for a local histopathologist that I decided to extend to
the List:

I have a client who is looking at deep-sea, meso- and surface soft coral
polyps, Gorgonians. She has contributed thousands of samples to (redacteD)
Museum, and they are having a strong disagreement about how they should be
stored. The old-timers say to store in 70% ethanol. I have more recently
heard arguments for 85% ethanol, but I can't document that. And she is
determined to store them in 95% ethanol, which sme museums seem to be
adopting. Certainly the protein and genomics look a lot better in meterial
stored in the 95% ethanol. Her mentor recently looked at polyps stored in
95% in one of those desktop SEMs, and he says he has less shriveling, but
we don't really have a good experimental set. While we wait to design
experiments to test this, I thought I would ask your opinion. What is the
current word on long-term storage for histopathology/museum specimens?

Meanwhile, she also had to collect some specimens into different
concentrations of rum, and now I want to test that, too, since it will
also have some sugars to keep the osmolarity up...!

Thoughts?

Aloha,
Tina

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

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Dear all,

We're about to move into a temporary facility, and before doing so had the
SEM vendor check the site to see if the vibration and electromagnetic
interference was within spec. The site has excess vibration from 0.1-3 Hz,
although it's not much over spec, but it was well out of spec for EMI. The
vendor recommended installing an active field cancelling system.

So, the architects got their own electromagnetic inspection and lo and
behold, they said the site was OK, so we're not getting a field cancelling
system. We're definitely having an inspection before re-installation of
the instrument, but I'm a bit stymied as to what we'll do if it's very
much out of spec.

Has this happened to anyone and how did you handle it?

thanks,
Rosemary

p.s. this email was rejected at first when in html format, not sure why.

Dr Rosemary White
CSIRO Black Mountain
GPO Box 1600
Canberra, ACT 2601
Australia

T 61 2 6246 5475
F 61 2 6246 5334
E rosemary.white-at-csiro.au

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Rosemary,

I expect to be in the same boat in a couple of years. Architects don't
like to listen to people who will be using their building.
I suggest pointing out to the money people that the EM vendors know
their instrument requirements and how to measure for them better than
anyone else. From experience. If the building doesn't meet their
criteria, then all performance warranties are void. If the instruments
can't work properly because of the building, then they are just
expensive paperweights.
Fixing the problem after the fact is *much* more expensive than doing
the job right in the first place.
You might also arrange a meeting between the vendor people and the
architect people to hash out the conflict. Both should want things to
work right.

Phil

} Dear all,
}
} We're about to move into a temporary facility, and before doing so had the
} SEM vendor check the site to see if the vibration and electromagnetic
} interference was within spec. The site has excess vibration from 0.1-3 Hz,
} although it's not much over spec, but it was well out of spec for EMI. The
} vendor recommended installing an active field cancelling system.
}
} So, the architects got their own electromagnetic inspection and lo and
} behold, they said the site was OK, so we're not getting a field cancelling
} system. We're definitely having an inspection before re-installation of
} the instrument, but I'm a bit stymied as to what we'll do if it's very
} much out of spec.
}
} Has this happened to anyone and how did you handle it?
}
} thanks,
} Rosemary
}
} p.s. this email was rejected at first when in html format, not sure why.
}
} Dr Rosemary White
} CSIRO Black Mountain
} GPO Box 1600
} Canberra, ACT 2601
} Australia
}
} T 61 2 6246 5475
} F 61 2 6246 5334
} E rosemary.white-at-csiro.au
--
Philip Oshel
Microscopy Facility Supervisor
Biology Department
024C Brooks Hall
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576

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Hello,
The electromagnetic interferences can change in magnitude! You have to carefully examine from which source the fields are produced. If they are, for instance, generated in an electric cable passing close to you SEM, and taking into account that the field strength depends on the current loaded on that cable, then you can easily imagine that if that cable serves in your building to power the air conditioning system, then you will observe very heavy and changing field strength over time. You will understand that the field might at certain times of the day be low, but at other times of the day produce unacceptable interferences. Something like this could explain why two professionals measured different field strength in your area.
We, encountering a similar situation some years ago, let a professional inspection of the electrical system in our building be done and found several violations of good practice of how to install power cables in a building. Correcting these reduced the electromagnetic interferences in the electron microscopy laboratories significantly - and we really have suffered very strong interferences!
While the electron microscopy vendor did an excellent job in detecting the presence of the fields, and the vendor, as well as some other companies, offered some nice solutions (active field cancellation systems, passive shielding), none of them had a solution for in detail locating the source of the problem and a solution to just remove that cause of the problem. As certified electricians are usually also not educated to conclude from a present field to the source of the problem, we have got the electrical system check in the building be done by a real professional on electromagnetic interferences, and the costs for his work and the corrections in the cabling (needed to be done some 20 meters away from the laboratory!) reduced the final total costs for the solution remarkably.

I that time have been in contact with many (internationally operating) shielding solution companies, from various continents, but in fact only one did care for the mayor cause of the problem and first solved that one before offering a final (then much smaller) shielding solution.
Although I am just a perfectly happy customer, I guess that I am not supposed to publish company names here on the microscopy-list, so contact me for further information if needed.

Wish you all the best,
with best greetings from the EM-Labs of CIC biomaGUNE,
Marco Möller

----------------------------------------------------------------
Marco Möller
Platform Manager - Electron Microscopy

CIC biomaGUNE
Parque Tecnológico de San Sebastián
Paseo Miramón, 182. Ed. Empresarial C
20009 San Sebastián (Guipúzcoa)
SPAIN

Tel: +34 943 00 53 25
mmoller-at-cicbiomagune.es

-----Original Message-----
X-from: Rosemary.White-at-csiro.au [mailto:Rosemary.White-at-csiro.au]
Sent: Wednesday, October 22, 2014 3:33 AM
To: Marco Moller

Dear all,

We're about to move into a temporary facility, and before doing so had the SEM vendor check the site to see if the vibration and electromagnetic interference was within spec. The site has excess vibration from 0.1-3 Hz, although it's not much over spec, but it was well out of spec for EMI. The vendor recommended installing an active field cancelling system.

So, the architects got their own electromagnetic inspection and lo and behold, they said the site was OK, so we're not getting a field cancelling system. We're definitely having an inspection before re-installation of the instrument, but I'm a bit stymied as to what we'll do if it's very much out of spec.

Has this happened to anyone and how did you handle it?

thanks,
Rosemary

p.s. this email was rejected at first when in html format, not sure why.

Dr Rosemary White
CSIRO Black Mountain
GPO Box 1600
Canberra, ACT 2601
Australia

T 61 2 6246 5475
F 61 2 6246 5334
E rosemary.white-at-csiro.au

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Tina,

For long-term storage ... more than one preservative.
Some samples in 95% for future protein/genomics work, some in 70% EtOH
(maybe 80%) + 5% glycerin for external morphology.

To be really exciting, I'd also process some samples for light or
confocal microscopy/TEM/SEM - fix/dehydrate/embed, and store as embedded
samples.
But long-term storage in alchohols is likely to lead to extracting
lipids and the morphology will Go Away.
Oh! Is there any possibility that the museum can store the samples below
25 deg C? 68 F. If yes, get them through 95% EtOH, then into 100%
t-butanol, let that freeze (25 C), and store them in that. Be better
than in EtOH of any %.
I assume t-BuOH is good for genomics, etc., but that should be checked,
and the morphology should be better. Decent for SEM, anyway.

Tissues in fixative are fine after a couple of years (even for TEM), but
... archival? Do you or the museum have some old samples that can be
sectioned and examined?

Phil

} I had a question for a local histopathologist that I decided to extend to
} the List:
}
} I have a client who is looking at deep-sea, meso- and surface soft coral
} polyps, Gorgonians. She has contributed thousands of samples to (redacteD)
} Museum, and they are having a strong disagreement about how they should be
} stored. The old-timers say to store in 70% ethanol. I have more recently
} heard arguments for 85% ethanol, but I can't document that. And she is
} determined to store them in 95% ethanol, which sme museums seem to be
} adopting. Certainly the protein and genomics look a lot better in meterial
} stored in the 95% ethanol. Her mentor recently looked at polyps stored in
} 95% in one of those desktop SEMs, and he says he has less shriveling, but
} we don't really have a good experimental set. While we wait to design
} experiments to test this, I thought I would ask your opinion. What is the
} current word on long-term storage for histopathology/museum specimens?
}
} Meanwhile, she also had to collect some specimens into different
} concentrations of rum, and now I want to test that, too, since it will
} also have some sugars to keep the osmolarity up...!
}
} Thoughts?
}
} Aloha,
} Tina
}
} ****************************************************************************
} * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu *
} * Biological Electron Microscope Facility * (808) 956-6251 *
} * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf*
} ****************************************************************************
--
Philip Oshel
Microscopy Facility Supervisor
Biology Department
024C Brooks Hall
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576

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Dear All, I would like to hear from you advices if one will prefably choose a TMP unit over a diffusion pump for the camera chamber. So far, i learn that TMP will take a shorter time to pump down the chamber. May i ask if there are many other advantages of using TMP over the diffusion pump?

Will it also reduce carbon contamination on the sample even though the sample should be in sample column area which is generally pumped down by an SIP.

Best Regards,
Yee Yan, Tay
FACTS Lab
Nanyang Technological University, Singapore

==============================Original Headers==============================
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Dear Tay Yee Yan,

If the turbo is backed by a scroll pump (dry): less hydrocarbon contamination, it is also better for any (cooled) cameras, you will need less heating cycles to keep the camera clean. In my experience any dry pump system will require less maintenance both on service and user level.

Best,

Wim Hagen
EMBL Heidelberg

} On Oct 22, 2014, at 18:42, rongchigram79-at-yahoo.com.sg wrote:
}
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} Dear All, I would like to hear from you advices if one will prefably choose a TMP unit over a diffusion pump for the camera chamber. So far, i learn that TMP will take a shorter time to pump down the chamber. May i ask if there are many other advantages of using TMP over the diffusion pump?
}
} Will it also reduce carbon contamination on the sample even though the sample should be in sample column area which is generally pumped down by an SIP.
}
} Best Regards,
} Yee Yan, Tay
} FACTS Lab
} Nanyang Technological University, Singapore
}
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6, 29 -- From wim.hagen-at-me.com Wed Oct 22 13:27:38 2014
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6, 29 -- Subject: Re: [Microscopy] Choosing a TMP Unit over a Diffusion Pump for the
6, 29 -- Camera Chamber in
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I am actually getting ready to replace the diff pump on my antique Hitachi S-450 SEM with a turbo.

Turbos have many advantages, no back streaming from the pump, you still may have back streaming from the roughing pump, though. Most modern turbo pumps are actually turbo-drag pumps, they have a molecular drag stage after the turbo stage which allows much higher backing pressures than a traditional turbo pump or a diffusion pump, this means you can get away with a cheaper rouging pump like a diaphragm pump or a scroll dry pump. Using these pumps mean there will be zero oil back streaming into the chamber.

Also turbo pumps spool up much faster than a diff pump warms up. The little pump I am putting on my SEM spins up in about 1 minute, the diff pump takes about 20 minutes to warm up. There is also probably no need for cooling on the turbo pump unless you are cycling the chamber a lot. Oh, and the working fluid in the diff pump is incredibly expensive for a SEM, most use Santovac 5 which is something like $250 for 100ml.

Now for the badâ€¦. They are kind of delicate critters, a friend at Portland State University has a Leybold maglev pump grenade on him the other day. Ripped it right off the mount. Donâ€™t know why it happened, must have been a failure in the maglev section which caused it to crash. Turbo pumps are much less tolerant of accidents than a diff pump, you let air into it and bad things happen with a turbo, with a diff pump you just get a mess.

There is also vibration, even the maglev pumps have some vibration. This is usually remedied with weighted isolation bellows.

-Jerry

} On Oct 22, 2014, at 9:45 AM, rongchigram79-at-yahoo.com.sg wrote:
}
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} Dear All, I would like to hear from you advices if one will prefably choose a TMP unit over a diffusion pump for the camera chamber. So far, i learn that TMP will take a shorter time to pump down the chamber. May i ask if there are many other advantages of using TMP over the diffusion pump?
}
} Will it also reduce carbon contamination on the sample even though the sample should be in sample column area which is generally pumped down by an SIP.
}
} Best Regards,
} Yee Yan, Tay
} FACTS Lab
} Nanyang Technological University, Singapore
}
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Hi Rosemary,

I don't know if you left out some details, but if it were me, I would not
accept just a statement from the architects. I would ask who they had do
the site survey, what are their qualifications, where and when were the
readings taken, and most of all, give me a copy of the report. I should
also point out that I would have watched the vendor do their site survey,
and required a copy of their report, also. I have learned not to flat out
accept this type of information, and to try to be a little bit scientific
in reviewing the information, especially if some would be selling me
something, or avoiding expense while providing something for me.

I wish you success!
Darrell

Rosemary.White-at-csiro.au wrote on 10/21/2014 09:18:58 PM:

} From: Rosemary.White-at-csiro.au
} To: Darrell Miles/Fishkill/IBM-at-IBMUS
} Date: 10/21/2014 09:19 PM
} Subject: [Microscopy] site inspections - overriding vendor
recommendations
}
}
}
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} Dear all,
}
} We're about to move into a temporary facility, and before doing so had
the
} SEM vendor check the site to see if the vibration and electromagnetic
} interference was within spec. The site has excess vibration from 0.1-3
Hz,
} although it's not much over spec, but it was well out of spec for EMI.
The
} vendor recommended installing an active field cancelling system.
}
} So, the architects got their own electromagnetic inspection and lo and
} behold, they said the site was OK, so we're not getting a field
cancelling
} system. We're definitely having an inspection before re-installation of
} the instrument, but I'm a bit stymied as to what we'll do if it's very
} much out of spec.
}
} Has this happened to anyone and how did you handle it?
}
} thanks,
} Rosemary
}
} p.s. this email was rejected at first when in html format, not sure why.
}
} Dr Rosemary White
} CSIRO Black Mountain
} GPO Box 1600
} Canberra, ACT 2601
} Australia
}
} T 61 2 6246 5475
} F 61 2 6246 5334
} E rosemary.white-at-csiro.au
}
}
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Dear Rosemary.

I confirm the many replies so far:

The vendor will not have to show any resolution if their measurement shows high fields, they will ask you to sign a waiver and any performance specs are out the door.

Try to get measurements over time, mornings, afternoons, evenings and also weekends:

I worked in a big city struggling with fields due to work on power lines two blocks down the road (TEM system in basement on corner of building), Friday afternoon it was gone forever.

A big EM vendor was about to spent a lot of money for a field cancelation/shielding solution in their factory when the problem was found to be wrongly connected room lighting on the other side of the building. This was found by the timing, 7am till 6pm during weekdays gave fields and this timing was traced to some room which was automatically powered down for safety reasons.

So it could be that there are fields only once and a while, differing in strength, try to figure that out and then possibly trace down where it comes from by talking to building maintenance as they have a good feel for what happens when in the building.
For the measurements it's best you get an independent expert, it will be worth the money. Alternatively ask your EM vendor to measure a few times during a full week. I think they should do that anyway, put in some black box that measures everything for a week and then have it analyzed by a professional.

Best,

Wim Hagen
EMBL Heidelberg

} On Oct 22, 2014, at 3:32, Rosemary.White-at-csiro.au wrote:
}
}
}
}
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
} To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
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} Dear all,
}
} We're about to move into a temporary facility, and before doing so had the
} SEM vendor check the site to see if the vibration and electromagnetic
} interference was within spec. The site has excess vibration from 0.1-3 Hz,
} although it's not much over spec, but it was well out of spec for EMI. The
} vendor recommended installing an active field cancelling system.
}
} So, the architects got their own electromagnetic inspection and lo and
} behold, they said the site was OK, so we're not getting a field cancelling
} system. We're definitely having an inspection before re-installation of
} the instrument, but I'm a bit stymied as to what we'll do if it's very
} much out of spec.
}
} Has this happened to anyone and how did you handle it?
}
} thanks,
} Rosemary
}
} p.s. this email was rejected at first when in html format, not sure why.
}
} Dr Rosemary White
} CSIRO Black Mountain
} GPO Box 1600
} Canberra, ACT 2601
} Australia
}
} T 61 2 6246 5475
} F 61 2 6246 5334
} E rosemary.white-at-csiro.au
}
}
}
} ==============================Original Headers==============================
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11, 28 -- From wim.hagen-at-me.com Wed Oct 22 15:09:34 2014
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I wouldn't put too much credence in the vendor's survey. The maintenance engineers have no special training or expertise in analyzing or mitigating these environmental problems. In our case, for TEM, it turns out the EMI from the power supply is well above the threshold that was imposed. I see no evidence that this affects our performance.

I don't recall having to sign a waiver, but I may have forgotten.
------------------------------------------
Phil Ahrenkiel

-----Original Message-----
X-from: wim.hagen-at-me.com [mailto:wim.hagen-at-me.com]
Sent: Wednesday, October 22, 2014 2:14 PM
To: Ahrenkiel, Phil

Dear Rosemary.

I confirm the many replies so far:

The vendor will not have to show any resolution if their measurement shows high fields, they will ask you to sign a waiver and any performance specs are out the door.

Try to get measurements over time, mornings, afternoons, evenings and also weekends:

I worked in a big city struggling with fields due to work on power lines two blocks down the road (TEM system in basement on corner of building), Friday afternoon it was gone forever.

A big EM vendor was about to spent a lot of money for a field cancelation/shielding solution in their factory when the problem was found to be wrongly connected room lighting on the other side of the building. This was found by the timing, 7am till 6pm during weekdays gave fields and this timing was traced to some room which was automatically powered down for safety reasons.

So it could be that there are fields only once and a while, differing in strength, try to figure that out and then possibly trace down where it comes from by talking to building maintenance as they have a good feel for what happens when in the building.
For the measurements it's best you get an independent expert, it will be worth the money. Alternatively ask your EM vendor to measure a few times during a full week. I think they should do that anyway, put in some black box that measures everything for a week and then have it analyzed by a professional.

Best,

Wim Hagen
EMBL Heidelberg

} On Oct 22, 2014, at 3:32, Rosemary.White-at-csiro.au wrote:
}
}
}
}
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
} To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
} On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} ----------------------------------------------------------------------------
}
} Dear all,
}
} We're about to move into a temporary facility, and before doing so had the
} SEM vendor check the site to see if the vibration and electromagnetic
} interference was within spec. The site has excess vibration from 0.1-3 Hz,
} although it's not much over spec, but it was well out of spec for EMI. The
} vendor recommended installing an active field cancelling system.
}
} So, the architects got their own electromagnetic inspection and lo and
} behold, they said the site was OK, so we're not getting a field cancelling
} system. We're definitely having an inspection before re-installation of
} the instrument, but I'm a bit stymied as to what we'll do if it's very
} much out of spec.
}
} Has this happened to anyone and how did you handle it?
}
} thanks,
} Rosemary
}
} p.s. this email was rejected at first when in html format, not sure why.
}
} Dr Rosemary White
} CSIRO Black Mountain
} GPO Box 1600
} Canberra, ACT 2601
} Australia
}
} T 61 2 6246 5475
} F 61 2 6246 5334
} E rosemary.white-at-csiro.au
}
}
}
} ==============================Original Headers==============================
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Hi

There are many advantages to using a turbo pump, but the main reason I do
not like them is the maintenance question. If you have a lab hundreds of
miles from your nearest maintenance technician, you are better off with a
diffusion pump. The great thing about a DP is that anyone with a mild degree
of skill can fix it when it stops working! I hate areas of a machine that
you cannot fix with over the phone help, when a turbo stops it stops, and
you have to wait for an exchange unit.

A tricky decision, good luck!

Steve

Steve Chapman FRMS
Protrain for Consultancy and Courses in Electron Microscopy
Mobile +44 (0) 7711 606967 Fax +44 (0)1280 814007
www.emcourses.com

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Title-Subject: [Filtered] postdoctoral researcher in customization of TEM

Message: We are seeking a collaborative researcher, possibly at a senior postdoctoral level, to
develop high precision, atomic-scale structural measurement techniques for the characterization of
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Email: michael_standing-at-byu.edu
Name: Michael Standing

Organization: Brigham Young University

Title-Subject: [Filtered] Textbook on biological EM specimen preparation

Message: Hello everyone,

We are in the process of developing a biological EM sample preparation
class. We haven't had this class available for a few years and the
previous instructor didn't use a formal text. We now are looking for a
textbook that will cover most of the basic preparation strategies for both
SEM and TEM biological sample preparations. Any and all recommendations
for texts that are currently in print would be very helpful for us.

Thanks in advance,

Mike
==============================
Michael D. Standing
BYU Microscopy Lab
108 MB
Provo, UT 84602

Phone: 801-422-4011
e-mail: michael_standing-at-byu.edu
==============================

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Name: Jo

Organization: University of Toledo

Title-Subject: [Filtered] High Pressure Freezing conditions for TEM

Message: Hello
I am trying to find the best condition for High pressure freezing and Freeze substitution methods
that will keep the samples in the most native as possible. More specifically, im wondering about the
filler/cryoprotectant usage for HPF, and ethanol or methanol for dehydration during Freeze
substitution.

thank you

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Hello Mike,

There is a nice and cheap book describing all TEM sample preparation in
a synthetic way and with the support of an interactive web site:

/Sample Preparation Handbook for Transmission Electron Microscopy/ from
Jeanne Ayache, Luc Beaunier, Jacqueline Boumendil, Gabrielle Ehret,
DaniÃ¨le Laub (vol1 /Methodology/, vol 2 /Techniques/).

The guide on the Web site can be run free, without the book, to choose a
method. The book brings more details on the procedures.

Have a look to
book: http://temsamprep.in2p3.fr/livre.php?lang=eng
web site: http://temsamprep.in2p3.fr/livre.php?lang=eng
basic guide: http://temsamprep.in2p3.fr/guides_methodologiques.php?lang=eng

Enjoy

Philippe Buffat

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Mike,

I'm hoping for good replies myself. We haven't found any recent EM texts
that cover biological prep for class use.
We're still using John Bozzola's book for our TEM course, hoping that
some day he'll get tired of retirement and write a new edition (you
still read this list, John?). The only other texts I know of are Glauert
& Lewis and Dystra and Reuss, but they're no newer.

There is a crying need for modern microscopy class texts, especially for
life sciences. The new EM books are excellent references, but not great
for class texts.

Phil

} Email: michael_standing-at-byu.edu
} Name: Michael Standing
}
} Organization: Brigham Young University
}
} Title-Subject: [Filtered] Textbook on biological EM specimen preparation
}
} Message: Hello everyone,
}
} We are in the process of developing a biological EM sample preparation
} class. We haven't had this class available for a few years and the
} previous instructor didn't use a formal text. We now are looking for a
} textbook that will cover most of the basic preparation strategies for both
} SEM and TEM biological sample preparations. Any and all recommendations
} for texts that are currently in print would be very helpful for us.
}
} Thanks in advance,
}
} Mike
} ==============================
} Michael D. Standing
} BYU Microscopy Lab
} 108 MB
} Provo, UT 84602
}
} Phone: 801-422-4011
} e-mail: michael_standing-at-byu.edu
} ==============================
--
Philip Oshel
Microscopy Facility Supervisor
Biology Department
024C Brooks Hall
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576

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Organization: University of Missouri

Title-Subject: [Filtered] CM10 and CM12 available

Message: We are looking for someone to take one or all of our three transmission electron
microscopes. We have two cm10 units and one cm12. One cm10 is not 100% complete and was used for
parts. The cm12 was up and running a few weeks ago before shutting it down. We are in the midst of
moving our lab off campus and do not have need for the scopes anymore.

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Listers,

As we all are aware, negative stains work by the fact that they consist of
heavier elements that scatter electrons better than the sample. They
surround the sample particles and pool in low areas or openings but
penetrate the sample themselves minimally in a well prepared sample.
However, some samples do much better than others in providing higher
contrast between the sample and background than others.

It is possible to get positive staining with a negative stain but this is
undesirable as it results in dark particles that tend to show little
detail or contrast against the background,.

In my experience, those particles that are of tighter construction such as
highly crystalline particles tend to take up less stain and thus appear
sharper and better defined than those that are more porous or have less
well defined edges in well-prepared negative stain preparations.

Do any of you know of any references that discuss negative staining and
the anticipated effect on crystalline verses non-crystalline particles?

Debby

Debra Sherman, Founder & CSO
DS imaging LLC
Purdue Technology Center
1281 Win Hentschel Blvd
West Lafayette, IN 47906
E-mail: debby.sherman-at-dsimagingllc.com
www.dsimagingllc.com
Mobile: 765-418-8540

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Hi, I am looking for a LEO 14xx SEM vacuum PCB and stage PCB boards for our
SEM. Could you please let me know if you happen to have one of these boards
for sale or if you know someone may have? Zeiss has the parts but won't sale
unless to be installed by them. Your help is greatly appreciated.

Thank you.
Ping

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Email: pedro.dacosta-at-kaust.edu.sa
Name: Pedro Costa

Organization: KAUST

Title-Subject: [Filtered] KAUST EM Symposium

Message: The King Abdullah University of Science and Technology (KAUST) is organizing its first
international symposium dedicated to electron microscopy, scheduled to take place from 9 to 11
December 2014. The event further celebrates the first five years of EM research activities at KAUST
as well as progress in the establishment of the EM field in the Middle East.

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It's no secret I'm a big, no a huge fan of the Wired website. They have a selection of winning images from the Nikon small world contest. As a Microscopist I'm fascinated both by the nature of the subject and the actual images as art.

John Delly use to tell me photomicrographs should be both beautiful and convey information to be great. These meet those criteria.

Here's a link,

http://www.wired.com/2014/10/nikon-small-world-40-years-microscope-photos/

Frank Karl

This email and any of its attachments may contain confidential information intended only for the use of the addressee(s). If the reader of this email is not the intended recipient or the employee or agent responsible for delivering it to the intended recipient, you are hereby notified that any dissemination or copying of this email is strictly prohibited. If you have received this email in error, please notify us by return email at info-at-ardl.com, permanently delete the email, and destroy any printouts. If this email contains test data and/or draft reports, you are hereby notified that only a signed original test report will contain official results, a copy of which resides in the project folder located at ARDL, Inc. Thank you. Akron Rubber Development Laboratory, Inc.

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Email: guosheng.liu-at-usask.ca
Name: Guosheng Liu

Organization: University of Saskatchewan

Title-Subject: [Filtered] Help for rotary microtome repair

Message: Hello Listers,

We inherited 4 ancient (1960's) rotary microtomes (LEITZ 1212 and SARTORIOUS) but none of them works
properly. After checkup, I found that all the problems were related to specimen feeding
sledge(reset)--either stuck due to long-time idle, or stuck due to the damaged thread (?). When
sledge moved to the limit, the user might have put extra force on the wheel hence damaged the threads.
My specific questions are: 1) is there any way to loosen the stuck sledge (apply special oil, or
organic solvent)? 2) are any special tools required to disassemble or assemble the parts? I found
the most screws were too tight to unscrew. Any other considerations when putting things back (like
calibrations, alignment)?
Thanks in advance.

Guosheng

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Guosheng,

Use light penetrating oil to free the stuck parts - let in soak in for
30 minutes to overnight.
And clean like mad. Most of the problem is going to be old dirt and old,
stiff lubricants.
No special tools are needed, just the usual screwdrivers, wrenches,
maybe a pliers, and something to throw in frustration.

If the feed threads are damaged, these can usually be fixed with a
thread-chasing tool or a thread tap, if one can found with the right
thread pitch.

Phil

} Email: guosheng.liu-at-usask.ca
} Name: Guosheng Liu
}
} Organization: University of Saskatchewan
}
} Title-Subject: [Filtered] Help for rotary microtome repair
}
} Message: Hello Listers,
}
} We inherited 4 ancient (1960's) rotary microtomes (LEITZ 1212 and SARTORIOUS) but none of them works
} properly. After checkup, I found that all the problems were related to specimen feeding
} sledge(reset)--either stuck due to long-time idle, or stuck due to the damaged thread (?). When
} sledge moved to the limit, the user might have put extra force on the wheel hence damaged the threads.
} My specific questions are: 1) is there any way to loosen the stuck sledge (apply special oil, or
} organic solvent)? 2) are any special tools required to disassemble or assemble the parts? I found
} the most screws were too tight to unscrew. Any other considerations when putting things back (like
} calibrations, alignment)?
} Thanks in advance.
}
} Guosheng
--
Philip Oshel
Microscopy Facility Supervisor
Biology Department
024C Brooks Hall
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576

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Best known to me commercially-available penetrant for loosing threads
and stuck gears on old instruments is "Kroil" from Kano Labs, but in the
pinch automotive break fluid and/or kerosene in combination with WD-40
would also do wonders. Wet generously and leave soaking overnight.

Valery Ray - also with NISP, UMDCP Nanocenter
=============================================
PBS&T, MEO Engineering Co., Inc.
290 Broadway, Suite 298
Methuen, MA 01844 USA
Phone: +1-978-296-5063 - leave a message with call-back number
US Mobile: +1-978-305-0479
Skype: pbstmeo
E-Mail: vray-at-partbeamsystech.com
UMD E-Mail: vray-at-umd.edu
Web: www.partbeamsystech.com
Web: www.freudlabs.com

On 10/29/2014 9:47 AM, microscopylistserver-noreply-at-microscopy.com wrote:
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} Email: guosheng.liu-at-usask.ca
} Name: Guosheng Liu
}
} Organization: University of Saskatchewan
}
} Title-Subject: [Filtered] Help for rotary microtome repair
}
} Message: Hello Listers,
}
} We inherited 4 ancient (1960's) rotary microtomes (LEITZ 1212 and SARTORIOUS) but none of them works
} properly. After checkup, I found that all the problems were related to specimen feeding
} sledge(reset)--either stuck due to long-time idle, or stuck due to the damaged thread (?). When
} sledge moved to the limit, the user might have put extra force on the wheel hence damaged the threads.
} My specific questions are: 1) is there any way to loosen the stuck sledge (apply special oil, or
} organic solvent)? 2) are any special tools required to disassemble or assemble the parts? I found
} the most screws were too tight to unscrew. Any other considerations when putting things back (like
} calibrations, alignment)?
} Thanks in advance.
}
} Guosheng
}
}
}
}
}
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Dear All,

does anybody know the name and function of the nodule-like structures on the bottom side of this sweet potato leaf:
http://www.electronmicroscopy.info/sweet_potato_leaf.jpg (3MB)
See detail in red circle.
There might also be some hyphen present...

Thanks,
Stefan

--

-----------------------------------------------------
Stefan Diller - Scientific Photography
Arndtstrasse 22
D - 97072 Wuerzburg Germany
++49-931-7848700 Phone
++49-931-7848701 Fax
++49-175-7177051 Mobile

Websites:
www.nanoflight.info
www.stefan-diller.com
www.electronmicroscopy.info
www.elektronenmikroskopie.info
www.zwillingsprojekt.de
www.assisi.de
Anfahrt: http://Mail.map24.com/Stefan.Diller
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Email: SciTech-at-wwu.edu
Name: Erin Macri

Organization: Western Washington University

Title-Subject: [Filtered] CPD-Liquid CO2 Consumption w/ E3000

Message: We have a Polaron E3000 series CPD and have been having ongoing trouble with liquid CO2
delivery to the chamber. Specifically, we have a great degree of variability in the number (1-8) of
CPD cycles we get from a full 50 lb cylinder with siphon tube. We do very little CPD work (2-3
users/year processing 50-100 samples) so I have had a difficult time determining the number of
cycles to expect from a cylinder. I chill the chamber to 10 C (with a recirculating chiller unit)
before starting and always do a test fill to check for leaks and liquid delivery before putting
samples in the chamber. We also weigh the cylinders to track usage and find that we still have 20 lb
or more left in the cylinder when it stops delivering liquid. I am aware that a full bottle
contains only ~42-43 lb of liquid and that the % liquid in the bottle decreases with increased room
temperature. Our lab is in the low to mid 70Â’s year-round. Just today my back-up cylinder stopped
delivering liquid after just one CPD cycle and my client lost 12 samples. We feel there may be a
supplier issue. We do tank exchanges so have a different cylinder each delivery.

My questions are:

How many cycles (initial fill and two flushes) do other users of this style CPD typically get from a
50 lb siphon bottle?
How much is left in the bottle once it no longer delivers liquid C02?
Has anyone experienced bad cylinders?

Thank you for your input!

Erin Macri
Instrument Center Services Coordinator
Scientific Technical Services
Western Washington University
Bellingham, WA 98225
www.wwu.edu/scitech/

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Nestor:

I am getting duplicate postings due to two TLDs being active.

gary-at-gaugler.com
gary-at-microtechnics.com

gary-at-gaugler.com should suffice. You can delete the microtechnics
address.

Tnx,
gary g.

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We've developed an unusual issue with our venerable (i.e., old) Noran Vantage EDS system. We are seeing an increase in background signal and a loss of peak intensity. The effect is especially pronounced in the low energy (mostly {2kV) portion of the spectrum where the background can be as much as 10 times its normal level. I can send you a file documenting the problem if you contact me directly.

We have also found that turning on the chamber scope and its IR illuminator seems to help temporarily restore functionality. First, the IR floods the detector with photons and the background control software complains and says it shuts off the bias to the detector. Once we turn off the IR and the control software recovers, we have maybe tens of seconds of fairly normal operation before the background problem gradually returns.

Is anyone familiar with this problem before? I've been using EDS since the late 1970s and have not seen this issue before.

Do you have any idea what to do about it - short of sending the detector in for evaluation?

Where may we find the bias value and/or setting within the Vantage software?

Thanks in advance.

Warren Straszheim (wearing my hat as Ames Lab's EPMA operator)

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Hi Warren

With the old liquid nitrogen cooled detectors a number of problems could
cause a drop off in low energy peak heights.

1) A degradation of the detector vacuum.
2) Icing up of the detector face due to holes in the window, also
contributing to the above.
3) A dirty window.

Others may know better, but my first step would be to disconnect the
detector from the rest of the system, and allow it to warn up. This would
remove any ice from the detector face. When cooling the detector back down,
and then running the system up again, if it takes a long time that would
indicate the vacuum problem. Most service technicians will have a device
that would allow a vacuum pump to interface with the detector.

Hope this helps?

Steve

Steve Chapman FRMS
Protrain for Consultancy and Courses in Electron Microscopy
Mobile +44 (0) 7711 606967 Fax +44 (0)1280 814007
www.emcourses.com

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Email: joseph.n.madary.civ-at-mail.mil
Name: NIck Madary

Organization: joint pathology center

Title-Subject: [Filtered] photo printer sugestions for TEM work?

Message: Hi all, We are using a Gatan Orius CCD attached to a JEOL 1400. We do mostly biologicals
and need the occasional high res print. We have not been able to get the best prints from what we
have now and would like to purchase something that just gives us a nice photo print without any
frills. No need for scanning, faxing, etc.

Does anyone have a newer printer they use, black and white they can recommend? Online is almost too
much info to decipher and I am tired of looking. I will now ask the experts! Thanks so much!

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Email: ravi.thakkar369-at-gmail.com
Name: Ravi

Title-Subject: [Filtered] Operation manual for sputter coater and CPD.

Message: Hi,
I would be great if you can send me user manual for
1. Sputter coater - Denton Vacuum Desk II
2. Critical Point Dryer - Samdri 790 B Tousimis

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Email: antoniom-at-ictan.csic.es
Name: Antonio D. Molina-García

Organization: Instituto de Ciencia y Tecnología de Alimentos y Nutrición-ICTAN (CSIC), Madrid, Spain

Title-Subject: [Filtered] Electronic boards for a Zeiss DSM 960 SEM

Message: Dear all,

We need some electronic boards for a Zeiss DSM 960 SEM. The equipment is old and we cannot find
any spares in Spain. We are looking for a “Video Mixer TV Generator 348332-9031 board”, a “Video
Switch 348332-9111 board” and a “BAS-interf 348332-9038 board”. The instrument is in a public
research centre in Madrid. We would be very grateful if someone can give as some clue of how to find
them.”

Best regards,

Antonio D. Molina-García
Instituto de Ciencia y Tecnología de Alimentos y Nutrición-ICTAN (CSIC)
José Antonio Nováis, 10
Ciudad Universitaria
28040 Madrid
SPAIN

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Hello Warren,
We had similar problem in our old EDAX DX4 system on Philips CM12/STEM
more then 10 years ago. I've got an advice through this listserver
from Matthew Ervin, which solve the problem.
I fish out the original message from Nestor's archive for you:
-------------------------------------------

Dear colleagues,

We have a position for an application engineer in electron microscopy in Frankfurt.

Details are below. For further information please contact me: achilleas.frangakis-at-biophysik.org

Best wishes,

Achilleas

The electron microscopy group at the Goethe University Frankfurt is seeking an application engineer in electron microscopy.

The job responsibilities include:

â€¢ Management and operation of the electron microscopes

â€¢ Recording of data for transmission and scanning electron microscopy.

â€¢ Training new users and providing support for the existing ones in using the EMs and the different software packages available.

â€¢ Trouble-shooting together with the FEI service engineers and Gatan support staff to assure minimal downtimes and optimal performance of the systems.

The position will be one of the staff positions in the electron microscopy group.

The successful applicant should have a solid background in the operation of latest generation electron microscopes and have a good understanding of high resolution imaging demands.

The applicant should be an enthusiastic and capable individual with excellent communication skills and committed to understanding and serving the user community.
--

Dr. Anja Seybert
Buchmann Institute for Molecular Life Sciences (BMLS)
Goethe University Frankfurt
Electron Microscopy
Frangakis Gruppe, EG 603, 1OG 1.658
Max-von-Laue-Str. 15
60438 Frankfurt am Main
Germany

Phone: +49-(0)69-798-42590
Fax: +49-(0)69-798-46467
seybert-at-biophysik.org
http://www.bmls.de/Electron_Microscopy/aboutus.html

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21, 20 -- "Microscopy microscopy.com" {Microscopy-at-microscopy.com}
21, 20 -- From: Anja Seybert {seybert-at-biophysik.org}
21, 20 -- Subject: Position for Application Engineer in Electron Microscopy in Frankfurt (Germany)
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Hi Warren and all

Good news to hear that there are still some other Vantages working
somewere !
Our one is still working too, and I'm not very in hurry to switch to a
newer system, in spite of the real necessity for a better detector.

I agree with Steve and Oldrich in their diagostic. We had 2 years ago a
similar problem, but with a very high deadtime (up to 80-90%) and a high
zero peak (PHA status/diagnostic) and no increase of the background
itself. And the same effect of the IR light, which reset for a short
time to better working conditons.

I warm up the detector, pump it down and bake it fresh (with the help
from someone of the list, which lend me the pumping coupling device).
Since it works well again. It has a noticable lost in resolution at low
energy but a correct sensitivity and no noise and deadtime any more.

The re-pumping and baking themself are not difficult if your are familar
with HV and UHV technology and if you have a turbo pump avaible. One
must just find the connecting device to open/close the pumping port and
pump down.

If your detector is 8-10 or more years old, and you warm it only up, you
may have the same problem again in one or two years. This was our case.
The windows is always a little bit porous and some air and water wapor
goes in the insulation vacuum with time. After 10 years the zeolites are
saturated. The warming up will remove the ice, but the zeolites will
saturate again very fast. A new pumping down with the baking of the
zeolites and all the internal surfaces of the cryostat (much much square
meters, with the super insulation film layers) is the best solution.

Hope it helps

Jacques

Le 30/10/2014 16:21, wesaia-at-iastate.edu a écrit :
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} We've developed an unusual issue with our venerable (i.e., old) Noran Vantage EDS system. We are seeing an increase in background signal and a loss of peak intensity. The effect is especially pronounced in the low energy (mostly {2kV) portion of the spectrum where the background can be as much as 10 times its normal level. I can send you a file documenting the problem if you contact me directly.
}
} We have also found that turning on the chamber scope and its IR illuminator seems to help temporarily restore functionality. First, the IR floods the detector with photons and the background control software complains and says it shuts off the bias to the detector. Once we turn off the IR and the control software recovers, we have maybe tens of seconds of fairly normal operation before the background problem gradually returns.
}
} Is anyone familiar with this problem before? I've been using EDS since the late 1970s and have not seen this issue before.
}
} Do you have any idea what to do about it - short of sending the detector in for evaluation?
}
} Where may we find the bias value and/or setting within the Vantage software?
}
} Thanks in advance.
}
} Warren Straszheim (wearing my hat as Ames Lab's EPMA operator)
}
}
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--

J. Faerber
IPCMS-DSI
Institut de Physique et Chimie des Matériaux de Strasbourg
Département Surfaces et Interfaces
23, rue de Loess ; BP43
67034 Strasbourg CEDEX 2
France

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

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X-from: a.helvoort-at-ntnu.no ()

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Email: a.helvoort-at-ntnu.no
Name: Ton van Helvoort

Organization: Norwegian University of Science and Technology (NTNU)

Title-Subject: [Filtered] Open PhD position FIB and TEM of semiconductor nanowire devices

Message: A PhD research fellowship is offered within the project Â“Low Cost, Ultra-High Efficiency
Graphene/Nanowire Solar CellsÂ”. The task of the announced PhD is to characterize the electronic
properties, structure and composition of the nanowire-devices by focused ion beam (FIB) and advance
transmission electron microscopy (TEM).
More details regarding the position and the application procedure can be found on:
http://www.ntnu.edu/geminicentre/tem/vacancies

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Dear Stefan,

Those could be glandular trichomes.
I could not find an example picture in the same species after a quick
internet search, but here is a paper with some of them in Ipomoea
cairica :

Paiva Elder AntÃ´nio Sousa, Martins Luiza Coutinho (2011) Calycinal
trichomes in Ipomoea cairica (Convolvulaceae): ontogenesis, structure
and functional aspects. Australian Journal of Botany 59, 91â€“98.

http://dx.doi.org/10.1071/BT10194

Cheers,

Rafael Buono

On Wed, Oct 29, 2014 at 11:11 AM, {stefan.diller-at-t-online.de} wrote:
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} Dear All,
}
} does anybody know the name and function of the nodule-like structures on the bottom side of this sweet potato leaf:
} http://www.electronmicroscopy.info/sweet_potato_leaf.jpg (3MB)
} See detail in red circle.
} There might also be some hyphen present...
}
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} Stefan
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8, 31 -- Subject: Re: [Microscopy] nodule-like structure on sweet potato leaf
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Hi,

We picked up this random leaf from the lawn and saw these "budding" structures on the leaf surface when doing cryoSEM on it. Does anyone have some idea what are these structures called on a leaf? Sorry, we don't know what kind of leaf it is when we pick it up.

http://emlab.uconn.edu/wp-content/uploads/sites/370/2014/11/Leaf-Surface.jpg

Xuanhao Sun, Ph.D.

Bioscience Electron Microscopy Laboratory
University of Connecticut
BPB G06, Unit 3242
Storrs, CT 06269-3242

Office Tel: 860-486-6368
Lab Tel: 860-486-2914
Fax: 860-486-6369
website: emlab.uconn.edu

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Email: michelle.gignac-at-duke.edu
Name: Michelle

Organization: Duke University

Title-Subject: [Filtered] Embedding Qustion

Message: Hello All.

I have a PI that is interested in immuno-labeling cells grown in 6 well plates. He would like to
keep the cells in the plates during dehydration and embedding to keep the cell-cell interactions
intact. The antigen he is interested in requires post embedding labeling. My question is what
resin can I use that will not interact with the plastic plates? Normally I would use the Polybed
812 resin, but will this work with immuno-labeling?

Thanks for your help.
Michelle

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Hi, Michelle:

It would be helpful if you can provide more information on the protein
your PI is trying to label and why he thinks the antigen requires post
embedding labeling. With all due respect to the users of our core, my
experience is most of them are not knowledgeable enough for immunoelectron
microscopy so although I listen to their opinion, I do have to make my own
decision on how to pursue to labeling. The important thing is to ask a lot
questions and to thoroughly understand the system/project. Feel free to
contact me off line if you want to discuss further. Thanks.

Hong
hyi-at-emory.edu

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Dear Xuanhao Sun,

these are multicellular trichomes that are quite common for plants.

Best regards,
Anne Heller

----- Nachricht von xuanhao.sun-at-uconn.edu ---------
Datum: Mon, 3 Nov 2014 16:04:47 -0600
Von: xuanhao.sun-at-uconn.edu
Antwort an: xuanhao.sun-at-uconn.edu
Betreff: [Microscopy] Help to identify a structure on leaf surface
An: Anne.Heller-at-uni-hohenheim.de

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} Hi,
}
} We picked up this random leaf from the lawn and saw these "budding"
} structures on the leaf surface when doing cryoSEM on it. Does anyone
} have some idea what are these structures called on a leaf? Sorry, we
} don't know what kind of leaf it is when we pick it up.
}
} http://emlab.uconn.edu/wp-content/uploads/sites/370/2014/11/Leaf-Surface.jpg
}
} Xuanhao Sun, Ph.D.
}
} Bioscience Electron Microscopy Laboratory
} University of Connecticut
} BPB G06, Unit 3242
} Storrs, CT 06269-3242
}
} Office Tel: 860-486-6368
} Lab Tel: 860-486-2914
} Fax: 860-486-6369
} website: emlab.uconn.edu
}
}
}
}
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} 7, 24 -- Subject: Help to identify a structure on leaf surface
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----- Ende der Nachricht von xuanhao.sun-at-uconn.edu -----

Dr. Anne Heller
AG Elektronenmikroskopie
Institut fÃ¼r Botanik (210)
UniversitÃ¤t Hohenheim
Postfach, 70593 Stuttgart
Tel. 0711-45922180
Fax. 0711-45923355

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Hello-

It's been a long time since I've done bio SEM prep, our lab mostly does
materials samples. But I've been getting more and more hydrated samples,
hydrogels, bacteria, etc. So far we've been air-drying, I do have access
to a freeze drier but haven't tried it yet. What is considered state-of
the art now for these kinds of preps? Critical point? Cryo-SEM? Fixation?

Thank you,
Leslie Thompson
IBM Research-Almaden
San Jose, CA

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3, 26 -- Subject: SEM sample prep for hydrogels?
3, 26 -- From: Leslie Thompson {2lesliethompson-at-gmail.com}
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X-from: lindamann1-at-yahoo.com ()

Leslie,

Air drying is the worst way to prepare hydrogels. The best way to do SEM
of these is cryoSEM: high-pressure freezing or plunge freezing into
slush nitrogen (not liquid nitrogen). Plunging into a LN2 cooled cryogen
is next best. Then image frozen - just vacuum sublimate the top
micrometer or so of the ice. Don't let the frozen gel get above -90 C.

Freeze-drying can work IFF it's done correctly - meaning stopping at
about -60 C and about -40 C to remove the structural water (aka "water
of collapse" for obvious reasons).

If you can't do either of those, CPD can work, but it will still alter
the gel structures.

One problem with CPD is the dehydration steps - those alter the gel
structure also. IFF your gels are of a nature that is not affected by
EtOH or acetone *and* not by t-butyl alcohol, you might try freeze
drying from t-butyl. Much simplier, but I don't know if anyone has
actually tried this.

But. Given these are hydrogels and the water is an important part of the
structure, cryoSEM is really the best and perhaps only way to image them
and get something close to reality.

Phil

} Hello-
}
} It's been a long time since I've done bio SEM prep, our lab mostly does
} materials samples. But I've been getting more and more hydrated samples,
} hydrogels, bacteria, etc. So far we've been air-drying, I do have access
} to a freeze drier but haven't tried it yet. What is considered state-of
} the art now for these kinds of preps? Critical point? Cryo-SEM? Fixation?
}
} Thank you,
} Leslie Thompson
} IBM Research-Almaden
} San Jose, CA
--
Philip Oshel
Microscopy Facility Supervisor
Biology Department
024C Brooks Hall
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576

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Name: Michael Cheatham

Organization: Syracuse University

Title-Subject: [Filtered] Troubleshootong a Gresham-Avalon-Spirit system

Message: My aplogies for using the web form. I am a subscriber to the Microscopy Listserv list but
I've never had luck posting directly.

I have just installed a new (to me) Gresham EDS detector (model 107-70, serial # 50798) onto the
fifth WDS position (most right hand side position) of a JEOL 8600. I acquired this detector a few
years ago from the Univ of Missouri Rolla campus. It was on a JEOL733 in the fourth position. I
used the same mounting plate to install it on my 8600. The Gresham is connected to a PGT Avalon
Model 8K box. I am using Spirit software version 1.7.5b.

In short, my problem is that I get no counts in the Spirit software.

The computer is a Dell with Windows XP. This system 9Dell/Spirit/Avalon/Gresham) was working in
Rolla. Since the computer came from the U of Missouri system I had to wipe it and re-install XP and
lost everything.

Might anyone on this list have a Gresham-Avalon-Spirit system? I need a few answers to help with
the complete software installation set-up.

I have a picture of the detector serial # plate and the interface to the 8600 column here:
http://earthsciences.syr.edu/Other/Gresham-EDS-SU.html

Can anyone help with the black knob at the base of the detector just at the interface plate. It has
six positions 0,1,2,3,4,5 abd I believe these select a mechanical aperture that rotates in front of
the end of the detector inside the vacuum chamber.

Mike

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Name: Rebecca Jackson

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Title-Subject: [Filtered] Digital Water Bath

Message: Morning all! We are looking at purchasing a water bath from Thermo Scientific. We always
had an analog one but we were thinking of purchasing a digital one with an internal thermometer.
Anyone have one? How long do the internal thermometers last? Any comments...?

Thanks!

-Bec.

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Email: DAVID.SMITH-at-asu.edu
Name: David Smith

Organization: Arizona State University

Title-Subject: [Filtered] Cluster hires in aberration-corrected electron microscopy

Message:
Department of Physics, Arizona State University, Tempe, AZ

The Department of Physics at Arizona State University (ASU) seeks to fill up to three
faculty positions in the topical area of aberration-corrected electron microscopy. Two
positions will be at the assistant/associate professor level and the third appointment is
an open rank position. Rank and tenure status will be commensurate with the
candidateÂ’s experience and publication record. Applicants must have a Ph.D. or
equivalent in physics or a closely related field at the time of appointment, and
demonstrated research experience in the field of aberration-corrected electron
microscopy. An outstanding record of accomplishments with clear potential to establish
a vigorous research program that will attract external funding, show a strong
commitment to excellence in teaching and demonstrated experience in a
multidisciplinary collaborative environment are highly desirable. The appointments are
expected to begin in August 2015.

We seek applicants who will help to leverage recent substantial investments in facilities
and instrumentation by the University for promoting cutting-edge electron microscopy.
ASU is home to the John M. Cowley Center for High Resolution Electron Microscopy
and has a well-established international reputation in transmission electron microscopy.
The university has recently established the Southwestern Center for Aberration
Corrected Electron Microscopy, which features three aberration-corrected instruments,
also allowing monochromated and in situ microscopy as well as electron holography.
The successful applicants are expected to develop strong experimental and/or
theoretical programs that will advance the scientific capabilities of these new
instruments.

Complete applications will include a cover letter, curriculum vitae, statements of plans
for research (3 to 5 pages) and teaching (1 page), and contact information for three
references (postal and email addresses, and phone number). Materials must be
submitted on-line at http://physics.asu.edu/employment.php. Only electronic
applications will be considered.

Review of applications will commence December 1, 2014. Further review of completed
files will be made every two weeks until the search is closed. A background check will
be required for employment.

Arizona State University is a VEVRAA Federal Contractor and an Equal
Opportunity/Affirmative Action Employer. All qualified applicants will be considered
without regard to race, color, sex, religion, national origin, disability, protected veteran
status, or any other basis protected by law.
https://www.asu.edu/aad/manuals/acd/acd401.html https://www.asu.edu/titleIX/

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South Dakota School of Mines and Technology has posted a job listing at the link below for a faculty position in the Nanoscience and Nanoengineering Ph.D. Program. Applicants should have research interest in advanced, optical bio-imaging methods:

http://www.nature.com/naturejobs/science/jobs/469827-assistant-associate-or-professor-nanoscience-and-nanoengineering

-Phil

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I have a Hitachi S-450 SEM that came with a Kevex EDS unit. The old model 7000 frame that came with it is dead and a friend gave me a "not quite as oldâ€ Kevex Sigma Gold MCA to control it. I was able to get service schematics and software for it from Thermo Fisher but I have not been able to scrounge up any manuals other than the basic operation manuals found on the web and on the CD with the software.

Connecting the Sigma to the sensor head does not looks to be a big deal, I just need to supply -24v for the 2003 preamp in place of the -15 used on the 2008 preamp. But where I am getting stuck is on the System Controller module, the schematics I was sent do not include this version of the module. The one on this rack has the two type E thermocouple inputs. What I donâ€™t know what to do with is the DB-15 for â€œBEAMâ€ and the MAP/SCAN connector. I have no idea how to set any of this up.

My unit also has the DBC card for connecting to the 4855 scope control box and would like to find any setup information on that unit as well, I still need to find one of those boxes, there is one on eBay for an almost reasonable price.

-Jerry

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Hello all,

My colleague and I are interested in compiling a list of all the
aberration corrected electron microscopes--SEM and TEM--in the US
(only in the US purely due to laziness and not patriotism). We
co-teach a course on electron microscopy and want to show how rapidly
these advanced instruments have propagated into universities and
industry. The early picture of AC-EM is easy to draw--there are just a
few data points to plot on our graphic--but the picture today is too
much work to compile for a two man team, yet we think it will drive
the point home to our students on how important this advancement in
electron optics is to the community.

Has anyone compiled such a list in the last year or two? If so, would
you be willing to share with us? Full credit will be given to you in
our lectures. Assuming such a list does not exist or is not recent,
please help us create one by providing the following information:

1) The name of your university or company if you have an aberration
corrected electron microscope
2) The number of aberration corrected microscopes in your facility
(please note if you have one of the very few AC-SEMs in the US;
otherwise S/TEM will be assumed)
3) Your city and state

The results will be overlaid on a map of the US, and we'll freely
share with the community.

Thank you for any help you can provide in compiling this list,
Chris
Senior Research Associate
Nanoscale Characterization and Fabrication Laboratory
Institute for Critical Technology and Applied Science
Virginia Tech
(540) 200-9511

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6, 26 -- Subject: List of aberration corrected electron microscopes in the US?
6, 26 -- From: Christopher Winkler {microwink-at-gmail.com}
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Looking for a vendor for 3 mm diameter TEM grids with a reference pattern
for identifying particular areas on the grid both microscopically and macroscopically.

Each individual hole must be between 5 to 10 microns, similar to 2000 mesh grid.

Krassimir Bozhilov.

==============================Original Headers==============================
3, 32 -- From krassimir.bozhilov-at-ucr.edu Fri Nov 7 18:20:46 2014
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3, 32 -- From: Krassimir Bozhilov {krassimir.bozhilov-at-ucr.edu}
3, 32 -- To: "microscopy-at-microscopy.com" {microscopy-at-microscopy.com}
3, 32 -- Subject: TEM reference grids
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Hi Krassimir,

I am not sure what is your particular TEM application, but if you can at
all use widely available 3mm TEM grids with SiN windows, then they are
fairly easy to mark or pattern on 5um to 10um scale with FIB...

Valery Ray - also with NISP, UMDCP Nanocenter
=============================================
PBS&T, MEO Engineering Co., Inc.
290 Broadway, Suite 298
Methuen, MA 01844 USA
Phone: +1-978-296-5063 - leave a message with call-back number
US Mobile: +1-978-305-0479
Skype: pbstmeo
E-Mail: vray-at-partbeamsystech.com
UMD E-Mail: vray-at-umd.edu
Web: www.partbeamsystech.com
Web: www.freudlabs.com

On 11/7/2014 7:22 PM, krassimir.bozhilov-at-ucr.edu wrote:
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} Looking for a vendor for 3 mm diameter TEM grids with a reference pattern
} for identifying particular areas on the grid both microscopically and macroscopically.
}
} Each individual hole must be between 5 to 10 microns, similar to 2000 mesh grid.
}
} Krassimir Bozhilov.
}
} ==============================Original Headers==============================
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} 3, 32 -- From: Krassimir Bozhilov {krassimir.bozhilov-at-ucr.edu}
} 3, 32 -- To: "microscopy-at-microscopy.com" {microscopy-at-microscopy.com}
} 3, 32 -- Subject: TEM reference grids
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4, 40 -- Date: Sat, 08 Nov 2014 14:15:09 -0500
4, 40 -- From: Valery Ray {vray-at-partbeamsystech.com}
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Hi Krassimir,

I am not sure what is your particular TEM application, but if you can at
all use widely available 3mm TEM grids with SiN windows, then they are
fairly easy to mark or pattern on 5um to 10um scale with FIB...

Valery Ray - also with NISP, UMDCP Nanocenter
=============================================
PBS&T, MEO Engineering Co., Inc.
290 Broadway, Suite 298
Methuen, MA 01844 USA
Phone: +1-978-296-5063 - leave a message with call-back number
US Mobile: +1-978-305-0479
Skype: pbstmeo
E-Mail: vray-at-partbeamsystech.com
UMD E-Mail: vray-at-umd.edu
Web: www.partbeamsystech.com
Web: www.freudlabs.com

On 11/7/2014 7:22 PM, krassimir.bozhilov-at-ucr.edu wrote:
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} Looking for a vendor for 3 mm diameter TEM grids with a reference pattern
} for identifying particular areas on the grid both microscopically and macroscopically.
}
} Each individual hole must be between 5 to 10 microns, similar to 2000 mesh grid.
}
} Krassimir Bozhilov.
}
} ==============================Original Headers==============================
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4, 40 -- Date: Sat, 08 Nov 2014 14:15:09 -0500
4, 40 -- From: Valery Ray {vray-at-partbeamsystech.com}
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Greetings Fellow Microscopists,

The Midwest Microscopy and Microanalysis Society will hold its final meeting of 2014 on Friday, November 21st, at Baxter Healthcare in Deerfield, IL. The topic is Multidimensional Microscopy - Correlative Approaches. We welcome MAS Tour Speaker Dr. Deborah Hall of Rush University Medical Center, and a host of other exciting speakers. A preliminary announcement can be found on the Meetings page of our website, and a final program will be posted soon:

http://www.midwestmicroscopy.org/

Please join us for an informative program and a chance to meet with local colleagues and vendors. We look forward to seeing you there!

Elaine Schumacher
M3S Program Coordinator

Elaine F. Schumacher | Senior Research Scientist
McCrone Associates, Inc. . 850 Pasquinelli Drive . Westmont, IL 60559
P. 630.887.7100 | F. 630.887.7417 | www.mccrone.com

==============================Original Headers==============================
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Hi all,
I have a antique gold wedding band(circa 1901) with an engraving on the
inside that is quite well worn, some parts barely readable under an
optical microscope. I mounted it in the SEM, with somewhat better
results, but am not able to decipher the entire engraving. Is there any
way I could enhance the engraving to make it more legible? I tried
wiping it colloidal graphite and then immediately wiping it out lightly
to see if the dag would settle in the depressions, but no luck. Thanks.

Gary

Gary M. Easton
Scanners Corporation
90 Aileron Court, Ste. 6
Westminster, Maryland USA 21157
410.857.7633 (Office)
443-524-9631(Fax)
717.634.2226 (Mobile)

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9, 17 -- To: "microscopy-at-microscopy.com" {microscopy-at-microscopy.com}
9, 17 -- Subject: Gold Ring engraving
9, 17 -- Date: Mon, 10 Nov 2014 16:47:04 +0000
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Hi Gary,

I do not know the answer, but metallography discussion board would seem
like a good place to look for engraving-in-metal enhancement techniques
and to post a question:

http://www.metallography.com/experts/forum.html

Valery Ray - also with NISP, UMDCP Nanocenter
=============================================
PBS&T, MEO Engineering Co., Inc.
290 Broadway, Suite 298
Methuen, MA 01844 USA
Phone: +1-978-296-5063 - leave a message with call-back number
US Mobile: +1-978-305-0479
Skype: pbstmeo
E-Mail: vray-at-partbeamsystech.com
UMD E-Mail: vray-at-umd.edu
Web: www.partbeamsystech.com
Web: www.freudlabs.com

On 11/10/2014 11:48 AM, garyeaston-at-scannerscorp.com wrote:
} ----------------------------------------------------------------------------
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} Hi all,
} I have a antique gold wedding band(circa 1901) with an engraving on the
} inside that is quite well worn, some parts barely readable under an
} optical microscope. I mounted it in the SEM, with somewhat better
} results, but am not able to decipher the entire engraving. Is there any
} way I could enhance the engraving to make it more legible? I tried
} wiping it colloidal graphite and then immediately wiping it out lightly
} to see if the dag would settle in the depressions, but no luck. Thanks.
}
} Gary
}
}
} Gary M. Easton
} Scanners Corporation
} 90 Aileron Court, Ste. 6
} Westminster, Maryland USA 21157
} 410.857.7633 (Office)
} 443-524-9631(Fax)
} 717.634.2226 (Mobile)
}
}
}
}
}
}
} ==============================Original Headers==============================
} 9, 17 -- From garyeaston-at-scannerscorp.com Mon Nov 10 10:47:12 2014
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} 9, 17 -- To: "microscopy-at-microscopy.com" {microscopy-at-microscopy.com}
} 9, 17 -- Subject: Gold Ring engraving
} 9, 17 -- Date: Mon, 10 Nov 2014 16:47:04 +0000
} 9, 17 -- Message-Id: {emf379f528-4c30-45bb-9ab2-ef701e92e7b8-at-garypc}
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Gary,

I'd try grazing-incidence light. Have the illumination parallel to the
metal surface. This will give the best chance of seeing surface topography.
In the SEM, have you tried Y-modulation scanning with BSE in the
topography mode?

Phil

} Hi all,
} I have a antique gold wedding band(circa 1901) with an engraving on the
} inside that is quite well worn, some parts barely readable under an
} optical microscope. I mounted it in the SEM, with somewhat better
} results, but am not able to decipher the entire engraving. Is there any
} way I could enhance the engraving to make it more legible? I tried
} wiping it colloidal graphite and then immediately wiping it out lightly
} to see if the dag would settle in the depressions, but no luck. Thanks.
}
} Gary
}
}
} Gary M. Easton
} Scanners Corporation
} 90 Aileron Court, Ste. 6
} Westminster, Maryland USA 21157
} 410.857.7633 (Office)
} 443-524-9631(Fax)
} 717.634.2226 (Mobile)
--
Philip Oshel
Microscopy Facility Supervisor
Biology Department
024C Brooks Hall
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576

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Gary,
Have you tried the side shot BSE using the SE detector? Just a thought.

One other thought: The engraving should disturb the crystal structure of
the gold, so it might be visible from an overhead BSE without etching the
gold, as long as the general surface hasn't been smeared. Might be worth a
try.
Ken

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

-----Original Message-----
X-from: garyeaston-at-scannerscorp.com [mailto:garyeaston-at-scannerscorp.com]
Sent: Monday, November 10, 2014 11:50 AM
To: kenconverse-at-qualityimages.biz

Hi all,
I have a antique gold wedding band(circa 1901) with an engraving on the
inside that is quite well worn, some parts barely readable under an
optical microscope. I mounted it in the SEM, with somewhat better
results, but am not able to decipher the entire engraving. Is there any
way I could enhance the engraving to make it more legible? I tried
wiping it colloidal graphite and then immediately wiping it out lightly
to see if the dag would settle in the depressions, but no luck. Thanks.

Gary

Gary M. Easton
Scanners Corporation
90 Aileron Court, Ste. 6
Westminster, Maryland USA 21157
410.857.7633 (Office)
443-524-9631(Fax)
717.634.2226 (Mobile)

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} We have a bad vibration problem in our building. Our old vibration pad has given up the ghost and is no longer made.

} Would like a recommendation for a anti vibration pad that can be up to four inches thick to place under the microtome to damp the vibrations, which are low frequency building vibrations…Many thanks..tom Reese, NIH

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Title-Subject: [Filtered] Tunable Light Source

Message: I am looking to talk to an expert in the field of microscopy, specifically microscopy lighting.

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Reese,
I've had some good success with Newport's "VIBe" series which are easily
configurable tables and they aren't as costly as the air based tables.
https://www.newport.com/VIBe-Mechanical-Vibration-Isolator-Platforms/987419/
1033/info.aspx

For a really inexpensive solution - you can try the Meade or Celestron
telescope vibration isolators - pack of 3 for about $50. One of my
customers used these under their Keyence digital scope and now they can get
good images at 2000X - they have a cooling tower with big fan motors right
outside on the roof near their lab.

Mike Toalson
Western Analytical Solutions, LLC
o • 208.899.4425 m • 208.340.3895
w • www.materialanalyzers.com e • mike-at-materialanalyzers.com

Representing:
HITACHI table-top SEM/EDS, LEICA Microsystems EM Sample Prep
(www.marinereef.com)
APPLIED RIGAKU TECHNOLOGIES XRF Elemental Analyzers
(www.rigakuedxrf.com)
RIGAKU RAMAN TECHNOLOGIES Raman Molecular Spectrometers
(www.rigakuraman.com)

-----Original Message-----
X-from: treese-at-mbl.edu [mailto:treese-at-mbl.edu]
Sent: Monday, November 10, 2014 1:23 PM
To: mike-at-materialanalyzers.com

} We have a bad vibration problem in our building. Our old vibration pad
has given up the ghost and is no longer made.

} Would like a recommendation for a anti vibration pad that can be up to
four inches thick to place under the microtome to damp the vibrations, which
are low frequency building vibrations…Many thanks..tom Reese, NIH

==============================Original Headers==============================
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15, 43 -- Subject: RE: [Microscopy] vibration table for Reichert
15, 43 -- Date: Mon, 10 Nov 2014 21:31:47 -0700
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Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

For another low-cost vibration-isolation solution, consider contacting
your local monument making vendor and orderring 12" thick granite plate
of suitable dimensions. To isolate granite place from the table
vibrations use tennis balls from your local sports supply store.
Quantity of tennis ball you will need depends on the mass of granite
plate and you instrument - try to have them compressed anywhere from 10%
to 25% of the tennis ball diameter.

Best of luck!

Valery Ray - also with NISP, UMDCP Nanocenter
=============================================
PBS&T, MEO Engineering Co., Inc.
290 Broadway, Suite 298
Methuen, MA 01844 USA
Phone: +1-978-296-5063 - leave a message with call-back number
US Mobile: +1-978-305-0479
Skype: pbstmeo
E-Mail: vray-at-partbeamsystech.com
UMD E-Mail: vray-at-umd.edu
Web: www.partbeamsystech.com
Web: www.freudlabs.com

On 11/10/2014 11:33 PM, mike-at-materialanalyzers.com wrote:
} ----------------------------------------------------------------------------
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} ----------------------------------------------------------------------------
}
} Reese,
} I've had some good success with Newport's "VIBe" series which are easily
} configurable tables and they aren't as costly as the air based tables.
} https://www.newport.com/VIBe-Mechanical-Vibration-Isolator-Platforms/987419/
} 1033/info.aspx
}
} For a really inexpensive solution - you can try the Meade or Celestron
} telescope vibration isolators - pack of 3 for about $50. One of my
} customers used these under their Keyence digital scope and now they can get
} good images at 2000X - they have a cooling tower with big fan motors right
} outside on the roof near their lab.
}
} Mike Toalson
} Western Analytical Solutions, LLC
} o • 208.899.4425 m • 208.340.3895
} w • www.materialanalyzers.com e • mike-at-materialanalyzers.com
}
} Representing:
} HITACHI table-top SEM/EDS, LEICA Microsystems EM Sample Prep
} (www.marinereef.com)
} APPLIED RIGAKU TECHNOLOGIES XRF Elemental Analyzers
} (www.rigakuedxrf.com)
} RIGAKU RAMAN TECHNOLOGIES Raman Molecular Spectrometers
} (www.rigakuraman.com)
}
}
} -----Original Message-----
} X-from: treese-at-mbl.edu [mailto:treese-at-mbl.edu]
} Sent: Monday, November 10, 2014 1:23 PM
} To: mike-at-materialanalyzers.com
} Subject: [Microscopy] vibration table for Reichert
}
}
}
}
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
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} ----------------------------------------------------------------------------
}
} } We have a bad vibration problem in our building. Our old vibration pad
} has given up the ghost and is no longer made.
}
} } Would like a recommendation for a anti vibration pad that can be up to
} four inches thick to place under the microtome to damp the vibrations, which
} are low frequency building vibrations…Many thanks..tom Reese, NIH
}
}
}
} ==============================Original Headers==============================
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} 15, 43 -- Subject: RE: [Microscopy] vibration table for Reichert
} 15, 43 -- Date: Mon, 10 Nov 2014 21:31:47 -0700
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Email: andel-at-cperi.certh.gr
Name: Andreas Delimitis

Organization: CPERI/CERTH

Title-Subject: [Filtered] SEM Wehnelt cup corrosion

Message: Dear listers,

I am contacting you regarding the JEOL 6400 SEM and, particularly, its Wehnelt unit.

On the three last times we exchanged the filament, we noticed that the Wehnelt cup at its outer
surface hole (where the edge of the W filament is located inside) appeared to be like corroded or
melt. The effect was more pronounced each time we removed the Wehnelt to exchange filament.
Furthermore, every new filament was placed into the centre of the cup hole and the gun operates just
below saturation conditions.

Any advice why this is happening and about possible ways to prevent it will be highly appreciated.

Thanks in advance,
Andreas.

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Andreas,
it seems you have flash-overs from the high voltage.
Check your vacuum in the gun section.
Do you have a stable imaging or does the image shift occasionally?

Best regards,
Stefan

Am 11.11.2014 14:01, schrieb microscopylistserver-noreply-at-microscopy.com:
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} Email: andel-at-cperi.certh.gr
} Name: Andreas Delimitis
}
} Organization: CPERI/CERTH
}
} Title-Subject: [Filtered] SEM Wehnelt cup corrosion
}
} Message: Dear listers,
}
} I am contacting you regarding the JEOL 6400 SEM and, particularly, its Wehnelt unit.
}
} On the three last times we exchanged the filament, we noticed that the Wehnelt cup at its outer
} surface hole (where the edge of the W filament is located inside) appeared to be like corroded or
} melt. The effect was more pronounced each time we removed the Wehnelt to exchange filament.
} Furthermore, every new filament was placed into the centre of the cup hole and the gun operates just
} below saturation conditions.
}
} Any advice why this is happening and about possible ways to prevent it will be highly appreciated.
}
} Thanks in advance,
} Andreas.
}
}
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--

-----------------------------------------------------
Stefan Diller - Scientific Photography
Arndtstrasse 22
D - 97072 Wuerzburg Germany
++49-931-7848700 Phone
++49-931-7848701 Fax
++49-175-7177051 Mobile

Websites:
www.nanoflight.info
www.electronmicroscopy.info
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www.assisi.de
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Anfahrt: http://Mail.map24.com/Stefan.Diller
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11, 23 -- From stefan.diller-at-t-online.de Tue Nov 11 07:55:53 2014
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Tom,

At the University of Pennsylvania I used a table constructed by using
cinderblocks for the support legs and a huge cement slab that I think is
commercially available as flooring in new buildings. This slab was
approximately 4 inches thick but maybe 6 feet long. Cement held the whole
thing together. A word of caution and that is to make sure that the table
does not touch any walls or the small vibrations that flow down the walls
will be transferred to your microtome.

Valery's suggestion has the potential of being nicer to look at than the
cement that I used and I have heard of using tennis balls previously but
have not tried them myself.

Opinions and experiences related are those of Pat Connelly and do not
represent the NIH. This message is not confidential and can be freely
shared and reproduced.

Pat

Patricia Stranen Connelly
Research Assistant
NHLBI EM Core Facility
National Institutes of Health
Building 14E Room 111B
Bethesda, MD 20892-5570
301-496-3491
connellyps-at-mail.nih.gov

http://www.nhlbi.nih.gov/research/intramural/researchers/core/electron-micr
oscopy-core/index.html

On 11/11/14 2:03 AM, "vray-at-partbeamsystech.com" {vray-at-partbeamsystech.com}
wrote:

}
} For another low-cost vibration-isolation solution, consider contacting
} your local monument making vendor and orderring 12" thick granite plate
} of suitable dimensions. To isolate granite place from the table
} vibrations use tennis balls from your local sports supply store.
} Quantity of tennis ball you will need depends on the mass of granite
} plate and you instrument - try to have them compressed anywhere from 10%
} to 25% of the tennis ball diameter.
}
} Best of luck!
}
} Valery Ray - also with NISP, UMDCP Nanocenter
} =============================================
} PBS&T, MEO Engineering Co., Inc.
} 290 Broadway, Suite 298
} Methuen, MA 01844 USA
} Phone: +1-978-296-5063 - leave a message with call-back number
} US Mobile: +1-978-305-0479
} Skype: pbstmeo
} E-Mail: vray-at-partbeamsystech.com
} UMD E-Mail: vray-at-umd.edu
} Web: www.partbeamsystech.com
} Web: www.freudlabs.com
}
} On 11/10/2014 11:33 PM, mike-at-materialanalyzers.com wrote:
} }
} } -------------------------------------------------------------------------
} } ---
} }
} } Reese,
} } I've had some good success with Newport's "VIBe" series which are easily
} } configurable tables and they aren't as costly as the air based tables.
} }
} } https://www.newport.com/VIBe-Mechanical-Vibration-Isolator-Platforms/9874
} } 19/
} } 1033/info.aspx
} }
} } For a really inexpensive solution - you can try the Meade or Celestron
} } telescope vibration isolators - pack of 3 for about $50. One of my
} } customers used these under their Keyence digital scope and now they can
} } get
} } good images at 2000X - they have a cooling tower with big fan motors
} } right
} } outside on the roof near their lab.
} }
} } Mike Toalson
} } Western Analytical Solutions, LLC
} } o € 208.899.4425 m € 208.340.3895
} } w € www.materialanalyzers.com e € mike-at-materialanalyzers.com
} }
} } Representing:
} } HITACHI table-top SEM/EDS, LEICA Microsystems EM Sample Prep
} } (www.marinereef.com)
} } APPLIED RIGAKU TECHNOLOGIES XRF Elemental Analyzers
} } (www.rigakuedxrf.com)
} } RIGAKU RAMAN TECHNOLOGIES Raman Molecular Spectrometers
} } (www.rigakuraman.com)
} }
} }
} } -----Original Message-----
} } X-from: treese-at-mbl.edu [mailto:treese-at-mbl.edu]
} } Sent: Monday, November 10, 2014 1:23 PM
} } To: mike-at-materialanalyzers.com
} } Subject: [Microscopy] vibration table for Reichert
} }
} }
} }
} } } We have a bad vibration problem in our building. Our old vibration pad
} } has given up the ghost and is no longer made.
} }
} } } Would like a recommendation for a anti vibration pad that can be up to
} } four inches thick to place under the microtome to damp the vibrations,
} } which
} } are low frequency building vibrationsŠMany thanks..tom Reese, NIH

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Andreas,
If what you're seeing looks like a peeling surface, then I think that you
are not cleaning all of the plated tungsten off of the interior of the
wehnelt. When it peels, it changes the electrostatic field and causes
imaging problems. Even if it doesn't peel, there is often gas trapped
beneath it that will eventually release in small pulses, making your beam
jump just a little each time. The trick is to inspect the wehnelt very
carefully after cleaning (with a light microscope) and reclean if necessary.

Ken Converse
owner

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

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Email: andel-at-cperi.certh.gr
Name: Andreas Delimitis

Organization: CPERI/CERTH

Title-Subject: [Filtered] SEM Wehnelt cup corrosion

Message: Dear listers,

I am contacting you regarding the JEOL 6400 SEM and, particularly, its
Wehnelt unit.

On the three last times we exchanged the filament, we noticed that the
Wehnelt cup at its outer
surface hole (where the edge of the W filament is located inside) appeared
to be like corroded or
melt. The effect was more pronounced each time we removed the Wehnelt to
exchange filament.
Furthermore, every new filament was placed into the centre of the cup hole
and the gun operates just
below saturation conditions.

Any advice why this is happening and about possible ways to prevent it will
be highly appreciated.

Thanks in advance,
Andreas.

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Dear All,
Sorry for disturbing.
For those who interested.
There is postdoc position for in situ TEM opening right now in argonne national lab. The basic request as following:
Considerable knowledge and hands on experience with in-situ analytical transmission electron microscopy and related techniques such as EELS and STEM.
Good knowledge and experience with x-ray fluorescence would be a plus.
Solid background in condensed matter physics, materials science and
catalysis.
Apply through: http://www.anl.gov/careers/apply-job/postdoctoral-applicants
(Requisition Number: 322846)
Thanks for your time.

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A friend wants to know:
Is there a commercial entity that will section TEM specimens (for pay) with a fast turnaround? She googled "commercial TEM sectioning" and came up with Agar scientific in the UK but would prefer a US company. I guess I should ask "how fast is fast?"

too slow,
Beth

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I posted a couple weeks back about a problem with a Noran Vantage Si(Li) detector. Those problems remain.

Many suggested the problem was due to ice, but I am reluctant to believe it. (You may check out an Excel file comparing various spectra or the EMSA files of the spectra at ftp://ftp.marl.iastate.edu/Equipment/Noran-EDS.)

Anyway, we know the vacuum is quite soft in the Dewar and we need to do something. If the sorbent is saturated and we warm the detector up to deal with ice or send it back for repair, we risk blowing out the window and incurring a more expensive repair.

Therefore, I wonder if we might beg, buy or borrow a fitting through which we could pump down the Dewar before and/or while we warm it up. It would be great if that alone solved the problem; however, I suspect we have other issues that will necessitate more serious refurbishing of the detector.

Warren Straszheim

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6, 32 -- From wesaia-at-iastate.edu Tue Nov 11 12:44:55 2014
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On Nov 11, 2014, at 10:59 AM, wesaia-at-iastate.edu wrote:

} I posted a couple weeks back about a problem with a Noran Vantage
} Si(Li) detector. Those problems remain.
}
} Many suggested the problem was due to ice, but I am reluctant to
} believe it. (You may check out an Excel file comparing various
} spectra or the EMSA files of the spectra at ftp://ftp.marl.iastate.edu/Equipment/Noran-EDS.)
}
} Anyway, we know the vacuum is quite soft in the Dewar and we need to
} do something. If the sorbent is saturated and we warm the detector
} up to deal with ice or send it back for repair, we risk blowing out
} the window and incurring a more expensive repair.
}
} Therefore, I wonder if we might beg, buy or borrow a fitting through
} which we could pump down the Dewar before and/or while we warm it
} up. It would be great if that alone solved the problem; however, I
} suspect we have other issues that will necessitate more serious
} refurbishing of the detector.
}
} Warren Straszheim

Dear Warren,
When we had this problem in Albany, our shop made a back plate out of
brass that had a copper tube with a Swagelok fitting on it. This
allowed us to attach the dewar to vacuum. We also from time to time
removed the zeolite and either regenerated it or replaced it. We
didn't have a mate to the existing vacuum fitting on the dewar, but
our work-around functioned very well.
Yours,
Bill

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Email: mclymer-at-datacolor.com
Name: Mark Clymer

Organization: Datacolor Inc.

Title-Subject: [Filtered] Nov 20 webinar: Â“Issues with Color in Histology in MedicineÂ” with Dr.
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Hi Gary

You can try to obtain a replica of your surface. RepliSet F5 designed by
Struers has an accuracy of 0.1µ and can be observed by SEM after coating.

Nicolas STEPHANT

Université de Nantes
Institut Jean Rouxel
Service de microscopie électronique à balayage et microanalyse
2 rue de la Houssinière
BP 92208
44322 Nantes cédex 3

Tél : 02 40 37 64 26
Mél : nicolas.stephant-at-univ-nantes.fr
Web : http://cnrs-imn.fr

"Le monde n'existe que pour autant que nous sommes capables d'en produire une image"
C.G Jung

Le 10/11/2014 17:58, garyeaston-at-scannerscorp.com a écrit :
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} Hi all,
} I have a antique gold wedding band(circa 1901) with an engraving on the
} inside that is quite well worn, some parts barely readable under an
} optical microscope. I mounted it in the SEM, with somewhat better
} results, but am not able to decipher the entire engraving. Is there any
} way I could enhance the engraving to make it more legible? I tried
} wiping it colloidal graphite and then immediately wiping it out lightly
} to see if the dag would settle in the depressions, but no luck. Thanks.
}
} Gary
}
}
} Gary M. Easton
} Scanners Corporation
} 90 Aileron Court, Ste. 6
} Westminster, Maryland USA 21157
} 410.857.7633 (Office)
} 443-524-9631(Fax)
} 717.634.2226 (Mobile)
}
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} 9, 17 -- From garyeaston-at-scannerscorp.com Mon Nov 10 10:47:12 2014
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} 9, 17 -- Subject: Gold Ring engraving
} 9, 17 -- Date: Mon, 10 Nov 2014 16:47:04 +0000
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8, 25 -- From Nicolas.Stephant-at-univ-nantes.fr Wed Nov 12 01:37:08 2014
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Hi John,
I cannot remember the set-up, since it has been well over a decade
since I was in Albany. Perhaps someone from Noran (or whichever company
bought them out) will be able to help.
Yours,
Bill

On Nov 11, 2014, at 5:23 PM, {jtwilley-at-sprynet.com} wrote:

} Bill,
}
} What is the procedure for removing the zeolite? I always assumed
} that it was somehow packaged inside the walls and therefore permanent.
}
} Best,
}
} John Twilley
} Consulting Scientist
}
}
} -----Original Message-----
} } From: wtivol-at-sbcglobal.net
} } Sent: Nov 11, 2014 4:29 PM
} } To: jtwilley-at-sprynet.com
} } Subject: [Microscopy] Re: Dewar vacuum connection
} }
} } On Nov 11, 2014, at 10:59 AM, wesaia-at-iastate.edu wrote:
} }
} } } I posted a couple weeks back about a problem with a Noran Vantage
} } } Si(Li) detector. Those problems remain.
} } }
} } } Many suggested the problem was due to ice, but I am reluctant to
} } } believe it. (You may check out an Excel file comparing various
} } } spectra or the EMSA files of the spectra at ftp://ftp.marl.iastate.edu/Equipment/Noran-EDS.)
} } }
} } } Anyway, we know the vacuum is quite soft in the Dewar and we need to
} } } do something. If the sorbent is saturated and we warm the detector
} } } up to deal with ice or send it back for repair, we risk blowing out
} } } the window and incurring a more expensive repair.
} } }
} } } Therefore, I wonder if we might beg, buy or borrow a fitting through
} } } which we could pump down the Dewar before and/or while we warm it
} } } up. It would be great if that alone solved the problem; however, I
} } } suspect we have other issues that will necessitate more serious
} } } refurbishing of the detector.
} } }
} } } Warren Straszheim
} }
} }
} } Dear Warren,
} } When we had this problem in Albany, our shop made a back plate out
} } of
} } brass that had a copper tube with a Swagelok fitting on it. This
} } allowed us to attach the dewar to vacuum. We also from time to time
} } removed the zeolite and either regenerated it or replaced it. We
} } didn't have a mate to the existing vacuum fitting on the dewar, but
} } our work-around functioned very well.
} } Yours,
} } Bill

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I would like to receive suggestions based on good experiences with a portable digital magnifier or microscope with the following capabilities:

Mag range: 5x-40x
Good depth of field
Flat field
lens magnification adjustment rather than digital
USB transfer of images to a tethered laptop or tablet, preferably in TIF
Narrow, wand-like profile
If lights are provided by LEDs, that they can be turned off to allow other light sources than axial.
Ability to adjust color temperature for alternate lighting conditions.

Please respond off-list. Thanks,

John Twilley
Dept. of Mat. Sci. and Eng.
Stony Brook, NY

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Hello Dear All

We are working on a Tungsten Jeol 5600LV and we consider to upgrade to the
LaB6 option, that will require the addition of an ion pump.
We would like to ask advice from you on the following:

-Is the quality of imaging of the LaB6 substantially better than the
Tungsten one?
-What would be the approximate cost to buy, install and maintain an ion pump
(maybe a used one)?
Vendors' responses welcome

Thanks
yorgos

Dr Yorgos Nikas
Athens Innovative Microscopy
Skra 36 Voula 16673 GREECE

Tel/fax +30 210 8957677
mobile +30 6945 107477
www.eikonika.net www.aim.cat
************************************

==============================Original Headers==============================
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8, 22 -- From: "yorgos nikas" {eikonika-at-otenet.gr}
8, 22 -- To: {microscopy-at-microscopy.com}
8, 22 -- Subject: Upgrade from Tungsten to LaB6. Is it worthy?
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Email: mckeown3-at-llnl.gov
Name: Joseph McKeown

Organization: LLNL

Title-Subject: [Filtered] Looking for Electronic boards for JEOL 2000FX

Message: Hi all,

We are looking for electronic boards for a JEOL 2000FX TEM, particularly the Vac System PB. We would
be very grateful if someone has spares they are willing to part with (we are willing to purchase
them) or has information on how to obtain them. Thanks!

Best regards,

Joe McKeown
Materials Science Division
Lawrence Livermore National Laboratory
Livermore, CA 94551
USA

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John,
Perhaps you could provide a summary to the list. Schools and science
oriented parents, among others, might be interested in what you learn.

Thanks

Ken Converse

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

-----Original Message-----
X-from: jtwilley-at-sprynet.com [mailto:jtwilley-at-sprynet.com]
Sent: Wednesday, November 12, 2014 9:53 PM
To: kenconverse-at-qualityimages.biz

I would like to receive suggestions based on good experiences with a
portable digital magnifier or microscope with the following capabilities:

Mag range: 5x-40x
Good depth of field
Flat field
lens magnification adjustment rather than digital
USB transfer of images to a tethered laptop or tablet, preferably in TIF
Narrow, wand-like profile
If lights are provided by LEDs, that they can be turned off to allow other
light sources than axial.
Ability to adjust color temperature for alternate lighting conditions.

Please respond off-list. Thanks,

John Twilley
Dept. of Mat. Sci. and Eng.
Stony Brook, NY

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Gary, Phil and others,

Well, we've moved. The SEM engineer seems reasonably happy with the new location - so far, but this may change when everything is switched on in all of the rooms. The larger problem was that the new area was "handed over" but not everything is finished - including gas lines to instruments, etc.

Thanks for all of your replies, very useful, though sadly we have not convinced the architects to change locations and dimensions of rooms in the new building to be completed in a couple of years. We will probably never be able to have a FESEM - even the architects' own EMI modellers said the interference would be quite high.

thanks,
Rosemary

Dr Rosemary White
CSIRO Black Mountain
GPO Box 1600
Canberra, ACT 2601

T 61 2 6246 5475

________________________________________
X-from: Gary Brown [microscopy.gmb-at-gmail.com]
Sent: Friday, 14 November 2014 6:25 a.m.
To: White, Rosemary (Agriculture, Black Mountain); Microscopy-at-microscopy.com
Cc: Gary Brown; oshel1pe-at-cmich.edu

Rosemary,

I expect to be in the same boat in a couple of years. Architects don't
like to listen to people who will be using their building.
I suggest pointing out to the money people that the EM vendors know
their instrument requirements and how to measure for them better than
anyone else. From experience. If the building doesn't meet their
criteria, then all performance warranties are void. If the instruments
can't work properly because of the building, then they are just
expensive paperweights.
Fixing the problem after the fact is *much* more expensive than doing
the job right in the first place.
You might also arrange a meeting between the vendor people and the
architect people to hash out the conflict. Both should want things to
work right.

Phil

} Dear all,
}
} We're about to move into a temporary facility, and before doing so had the
} SEM vendor check the site to see if the vibration and electromagnetic
} interference was within spec. The site has excess vibration from 0.1-3 Hz,
} although it's not much over spec, but it was well out of spec for EMI. The
} vendor recommended installing an active field cancelling system.
}
} So, the architects got their own electromagnetic inspection and lo and
} behold, they said the site was OK, so we're not getting a field cancelling
} system. We're definitely having an inspection before re-installation of
} the instrument, but I'm a bit stymied as to what we'll do if it's very
} much out of spec.
}
} Has this happened to anyone and how did you handle it?
}
} thanks,
} Rosemary
}
} p.s. this email was rejected at first when in html format, not sure why.
}
} Dr Rosemary White
} CSIRO Black Mountain
} GPO Box 1600
} Canberra, ACT 2601
} Australia
}
} T 61 2 6246 5475
} F 61 2 6246 5334
} E rosemary.white-at-csiro.au
--
Philip Oshel
Microscopy Facility Supervisor
Biology Department
024C Brooks Hall
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576 {tel:%28989%29%20774-3576}

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Good Afternoon Dr. McKeown,

We are the long-time hosts to an instrument (JEM-4000EX UHV TEM) which was received from Xerox Corp as an instrument donation to UW-Madison, Materials Science and Engineering where it languished on a loading dock for several years before it was shipped to us at UW-Milwaukee, where we began the assembly - gun and HV tank have never been mounted. It has never been leak tested; the trials involved trying to pump the system down without the gun mounted, but with the UHV/FIB stage mounted. It was powered up once with the JEOL engineer on hand, but there was an electric discharge and the power was cut. The engineer checked over several boards and made some repairs. But it never has been powered up since. There it sits, forlorn and silent, with all the electronic equipment (computer work stations from 1990's). We think all the parts are here, but when it shipped from Madison we suspected there might have been a crate or two missing, but could not identify which parts.

If you are still interested or know of anyone who might be, to host this instrument or perhaps parts thereof, please reply with contact particulars.

Do you suppose you might be able to use some of the boards from this instrument?

Respectfully Yours,

Donald Robertson
Sr. Instrumentation Specialist
HRTEM Lab / Lab for Surface Studies
Department of Physics
College of Letters and Science / Graduate School
University of Wisconsin - Milwaukee
1900 East Kenwood Boulevard
Milwaukee, Wisconsin 53211
Office: Physics 329a
Telephone: (414) 229 2753
Email: donald-at-uwm.edu

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9, 37 -- 2000FX
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Yorgos,

My experience is limited to one ISI DS-130 so it may not be useful to you, or widely applicable.

The DS-130 had a LaB6 filament when I came to work here, and I dutifully tended the ion pump, worried about it when the power went out, and replaced the filament every few years. I generally got over 1,800 hours use from a filament, either Denka or Kimball Physics.

When circumstances (long-term power failure consequent to a Gulf storm) eventually killed the ion pump, I switched to tungsten "until new grids for the ion pump come in" - and was totally satisfied with the imaging. For EDS (materials / metallurgy application) the X-ray count rate with LaB6 is better, but adequate with tungsten.

Al low accelerating voltages (e.g 3-5 KV - I look at explosives powders as well as metals), the LaB6 is brighter/superior. At 15/20 KV tungsten is entirely satisfactory.

I quit worry about the ion pump and about filament life (I get close to 80 hours average on tungsten - book says 50 is usual) - but I let the gun cool for a while before specimen exchanges, slows me down but gentle on the microscope/filament.

For most uses the tungsten filament is satisfactory and I have no urge to re-install the LaB6 filament and refurbished ion pump.

I hope this helps. Others with more and wider experience will likely come by with other opinions.

Regards,
Andrew Werner
Chief Metallurgist, Perforating
Schlumberger Reservoir Completions
14910 Airline Road
Rosharon, TX 77583

-----Original Message-----
X-from: eikonika-at-otenet.gr [mailto:eikonika-at-otenet.gr]
Sent: Thursday, November 13, 2014 1:31 AM
To: Andrew Werner

Hello Dear All

We are working on a Tungsten Jeol 5600LV and we consider to upgrade to the
LaB6 option, that will require the addition of an ion pump.
We would like to ask advice from you on the following:

-Is the quality of imaging of the LaB6 substantially better than the
Tungsten one?
-What would be the approximate cost to buy, install and maintain an ion pump
(maybe a used one)?
Vendors' responses welcome

Thanks
yorgos

Dr Yorgos Nikas
Athens Innovative Microscopy
Skra 36 Voula 16673 GREECE

Tel/fax +30 210 8957677
mobile +30 6945 107477
www.eikonika.net www.aim.cat
************************************

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On Nov 12, 2014, at 11:33 PM, eikonika-at-otenet.gr wrote:

} Hello Dear All
}
} We are working on a Tungsten Jeol 5600LV and we consider to upgrade
} to the
} LaB6 option, that will require the addition of an ion pump.
} We would like to ask advice from you on the following:
}
} -Is the quality of imaging of the LaB6 substantially better than the
} Tungsten one?
} -What would be the approximate cost to buy, install and maintain an
} ion pump
} (maybe a used one)?
} Vendors' responses welcome
}
} Thanks
} yorgos
}
}
} Dr Yorgos Nikas
} Athens Innovative Microscopy
} Skra 36 Voula 16673 GREECE

Dear Yorgos,
LaB6 is a brighter and more coherent source than W. The filaments
are more expensive than W, but they last considerably longer if
treated well. Whether you will find the quality of your images to be
sufficiently better depends on what you plan to image.
Yours,
Bill

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Dear Friends
Thanks a lot for your answers. It looks like LaB6 is beautiful but, as
usual, this luxury comes with expenses and difficulties. Therefore I may
have to abandon this idea (with the mixed feelings of someone who decides to
follow the Prudence and not the Beauty..).
Best regards
yorgos

At 11:23 PM 11/12/2014, you wrote:

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Hello Dear All

We are working on a Tungsten Jeol 5600LV and we consider to upgrade to the
LaB6 option, that will require the addition of an ion pump.
We would like to ask advice from you on the following:

-Is the quality of imaging of the LaB6 substantially better than the
Tungsten one?
-What would be the approximate cost to buy, install and maintain an ion
pump
(maybe a used one)?
Vendors' responses welcome

Thanks
yorgos

Dr Yorgos Nikas
Athens Innovative Microscopy
Skra 36 Voula 16673 GREECE

Tel/fax +30 210 8957677
mobile +30 6945 107477
www.eikonika.net www.aim.cat
************************************

==============================Original Headers==============================
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We at the University of Illinois, would like to congratulate the winners of our first ever Fall Biological Conference poster and presentation winners!

This Contest was advertised on the microscopy listserve last summer.

The Zeiss award for the best poster: Elizabeth Joachim

Gelatin Nanoparticles for Non-invasive Delivery of Neuroprotectants

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

The Asylum Research (an Oxford Company) award for the best speaker presentation: Sage Dunham

Architectural and Chemical Characterization of Bacterial Biofilm with Secondary Ion Mass Spectrometry and Electron Microscopy

We congratulate these students, on this fine achievement. And hope to carry on the tradition of this competition.

Lou Ann Miller

{ { { { { { { { { { { { { { { { {} } } } } } } } } } } } } } } }
Lou Ann Miller
Biological Electron Microscopy
Frederick Seitz Materials Research Laboratory
Lab Room 125 Office 374 MRL
104 South Goodwin Ave
Urbana, IL 61801
Campus mail code: MC-230

217-244-1567
lamiller-at-illinois.edu
http://mrl.illinois.edu

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Email: vakimler-at-med.wayne.edu
Name: Vickie Kimler

Organization: Wayne State University

Title-Subject: [Filtered] Uneven Embed 812 Polymerization

Message: Hello All,
How can I correct the uneven polymerization of my resin in conical tip
"bottleneck" Beem capsules?

The tips where my pelleted cells are situated, are still soft, while the
top of the block (base) is very hard.

I know my mistake that I did not use a metal heat sink on the aspect of
the tips.

Can I continue to polymerize them in the 70 degree C oven with a heat
sink without destroying interrelationships between organelles? They have
been out of the oven a few days now, of course.

Any other advice?
Thanks,
Vickie

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Hi all,

we would like to find out where exactly the changes take place in the column when the probe current setting is altered between HIGH and NORM.

We are trying to understand why NORM setting gives higher resolution.

Regards,

Bettina and Sascha

________________________________________________________________
Â
Dipl. Ing. (FH) Bettina Wolpensinger
Â
Abteilung Photovoltaik
Photovoltaics Department
Â
Elektronenmikroskopie
Electron Microscopy
Institut fÃ¼r Solarenergieforschung GmbH
Am Ohrberg 1
D-31860 Emmerthal
Phone: +49 (0) 5151 999 313
(BÃ¼ro) oder 218 (Mikroskop)
Fax: +49 (0) 5151 999 400
eMail: b.wolpensinger-at-isfh.de
Internet: www.isfh.de
Handelsregister: Amtsgericht Hannover HRB 100547
GeschÃ¤ftsfÃ¼hrer: Prof. Dr. Rolf Brendel
Vorsitzender des Aufsichtsrats: Ministerialrat Dr. Hans Schroeder
________________________________________________________________
Â
Â
Â

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Hello All,

The University of Florida has posted an EM technician job.

The Electron Microscopy Core Laboratory of the Interdisciplinary Center
for Biotechnology Research (ICBR) at the University of Florida is looking
for an electron microscopist to join our team. This unique career
opportunity will allow you to apply and enhance your technical skills and
scientific knowledge in a friendly environment. This is a full-time,
time-limited position.

Visit the MSA job placement site; Job ID 21238688
http://jobs.microscopy.org/jobseeker/job/21238688/
or go to https://jobs.ufl.edu/postings/58970.

Karen Kelley

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Which Hitachi FESEM do you have? If the controls are labelled the same
as on the other Hitachi's I've used, including FESEM, the probe current
actually focuses the condenser len(s). The result of this is to change
the spot size of the probe where it impacts the sample. Higher probe
current means a larger spot size which means lower resolution.
High probe currents are typically used for backscattered imaging and
x-ray analysis (and low vacuum imaging, if your FESEM has that), where
resolution is traded away to get more signal and a better signal to
noise ratio.

Phil

} Hi all,
}
} we would like to find out where exactly the changes take place in the column when the probe current setting is altered between HIGH and NORM.
}
} We are trying to understand why NORM setting gives higher resolution.
}
} Regards,
}
} Bettina and Sascha
}
}
} ________________________________________________________________
} Â
} Dipl. Ing. (FH) Bettina Wolpensinger
} Â
} Abteilung Photovoltaik
} Photovoltaics Department
} Â
} Elektronenmikroskopie
} Electron Microscopy
} Institut fÃ¼r Solarenergieforschung GmbH
} Am Ohrberg 1
} D-31860 Emmerthal
} Phone: +49 (0) 5151 999 313
} (BÃ¼ro) oder 218 (Mikroskop)
} Fax: +49 (0) 5151 999 400
} eMail: b.wolpensinger-at-isfh.de
} Internet: www.isfh.de
} Handelsregister: Amtsgericht Hannover HRB 100547
} GeschÃ¤ftsfÃ¼hrer: Prof. Dr. Rolf Brendel
} Vorsitzender des Aufsichtsrats: Ministerialrat Dr. Hans Schroeder
--
Philip Oshel
Microscopy Facility Supervisor
Biology Department
024C Brooks Hall
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576

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Dear Vickie,
You may have a closer look to Kent McDonald's paper in Protoplasma (2014)
251:429â€“448: he even suggested 100 C (!!!) for Epon + LRW, and did so , with
all kinds of biological samples, with great success. No BEEM, though, if I
remember correctly.
Kind regards! Reinhard

--
Prof. Dr. Reinhard Rachel
University of Regensburg
Centre for EM / Anatomy
Faculty of Biology & Preclin. Med.
Universitaetsstrasse 31
D-93053 Regensburg - Germany
tel +49 941 943 -2837, -1720
mail reinhard.rachel-at-biologie.uni-regensburg.de
office: VKL 3.1.29

Next microscopy conferences:
http://www.mc2015.de
DGE - Microscopy Conference 2015, Sept 6-11, GÃ¶ttingen
http://www.emc2016.fr/en/
16th Europ Microsc Congress EMC 2016 in Lyon, FR
next Microbiol. conferences:
VAAM - Annual Conf March 1-4, 2015, Marburg

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HiÂ

â€ŽIf your instrument is configured the same as the Hitachi S-4800, the HIGH/NORM toggle changes C1. Â â€Ž

Under Setup } Column you can adjust C1 excitation between 1 and 16, where 1 is lowest excitation (largest probe) and 16 is highest excitation (smallest probe), but you can change the offset of this with NORM/HIGH. Â

When you change from NORM to HIGH you are effectively changing excitation on C1 to a new range. HIGH-1 is the least excitation possible (biggest probe available), down to HIGH-16, then NORM-1 is next, down to NORM-16 (most excitation available, smallest probe).

If you plot specimen current (put a Faraday cup on the stage and connect a picoammeter to the stage BNC port) as a function of C1 and NORM/HIGH, you'll be able to see the relationship.

HTH,

--â€Ž
Adam

Â Original Message Â
X-from: wolpensinger-at-isfh.de
Sent: Tuesday, November 18, 2014 3:34 AM
To: adamschuetze-at-outlook.com
Reply To: wolpensinger-at-isfh.de

Hi all,

we would like to find out where exactly the changes take place in the column when the probe current setting is altered between HIGH and NORM.

We are trying to understand why NORM setting gives higher resolution.

Regards,

Bettina and Sascha

________________________________________________________________
Ã‚Â
Dipl. Ing. (FH) Bettina Wolpensinger
Ã‚Â
Abteilung Photovoltaik
Photovoltaics Department
Ã‚Â
Elektronenmikroskopie
Electron Microscopy
Institut fÃƒÂ¼r Solarenergieforschung GmbH
Am Ohrberg 1
D-31860 Emmerthal
Phone: +49 (0) 5151 999 313
(BÃƒÂ¼ro) oder 218 (Mikroskop)
Fax: +49 (0) 5151 999 400
eMail: b.wolpensinger-at-isfh.de
Internet: www.isfh.de
Handelsregister: Amtsgericht Hannover HRB 100547
GeschÃƒÂ¤ftsfÃƒÂ¼hrer: Prof. Dr. Rolf Brendel
Vorsitzender des Aufsichtsrats: Ministerialrat Dr. Hans Schroeder
________________________________________________________________
Ã‚Â
Ã‚Â
Ã‚Â

Â
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Email: 13qw9-at-queensu.ca
Name: Jason

Organization: Queen's University

Title-Subject: [Filtered] what's the difference between mapping,
Hypermap and QMap in Esprit?

Message: hi all, are there anybody using Bruker Esprit for elemental
mapping in TEM? Could you please inspire me a bit about what's the
difference between Mapping, HyperMap and QMap? Which one gives the real
elemental content and distribution image? What I found is, after
processing the Hypermaps using QMap, many tiny but shiny spots come out
which actually don't make sense to me and the material. Thank you!

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Dear Jason,

When you take a HyperMap you save the whole EDX spectrum for each spatial pixel x, y to record the full spectal data cube. This allows you to process the data cube after it has been recorded, for instance creating maps from elements which you would not originally have chosen, pulling out line-scans or running the QMap (see later). If you instead take a Mapping you record only the integrated intensity for each chosen EDX peak, in other words acquiring a series of 2D map images rather than the full 3D data cube. Because you have to choose the elements to map before you acquire, and because you loose the spectral information which is important for checking that your maps are correct (for instance in STEM-EDX mapping it is easy to make fictive maps of background), and because you cannot do post-processing, I would never recommend this. Always record the HyperMap.

Once you have acquired the HyperMap you can then run a quantification (e.g. wt.% or at.%) of the data cube for chosen elements using the QMap function. Note that you can include elements for deconvolution without quantifying them, for instance to account for Cu signal from the grid. Whether or not the quantified maps give the correct elemental distribution depends on your specimen and on your point of view. If your specimen is a uniformly thick lamella and if you have enough counts per pixel that each pixel has a good quality spectrum then it should be good. However, if you have particles sitting on a carbon film or other objects of varying thickness then maybe not. This is for three reasons. First by calculating percentage composition you flatten the increase in elemental intensity that you associate with the object becoming thicker. Secondly small particles or objects may never give enough counts per pixel to give a spectrum that can be properly fitted and quantified (a situation that can sometimes be remediated by running a binned QMap). Thirdly the QMap will quantify every pixel, including for instance the carbon film and pixels in the vacuum. The latter two aspects differentiate STEM-EDX mapping from SEM-EDX mapping, which is typically done on bulk samples. They further have the effect of amplifying noise, since essentially you run noise peaks through a quantification. This probably accounts for the shiny spots that you observe in your QMaps. To check for this, use the spectrum picker tool to look at the quality of the spectra for individual pixels and see if it is reasonable to fit a background and quantify them or if each spectrum actually has too few counts.

As a final note, a “middle way” exists between the raw elemental maps – which are not background subtracted and which scale intensities between zero and maximum counts irrespective of whether there is 0.1 at.% or 99.9 at.% of an element – and the wt.% or at.% maps which have the drawbacks discussed above. This is to run the QMap and then ask to display the deconvolved, integrated peak intensities. If the data are noisy you can then follow this with a pixel smoothing. The resultant maps can give more truly comparative intensities while avoiding the “flattening” of intensity changes that are related to thickness changes.

Regards
Duncan – working with the Super-X and Bruker Esprit since 2012...

On 18 Nov 2014, at 18:20, {zaluzec-at-microscopy.com} {zaluzec-at-microscopy.com} wrote:

} Email: 13qw9-at-queensu.ca
} Name: Jason
}
} Organization: Queen's University
}
} Title-Subject: [Filtered] what's the difference between mapping,
} Hypermap and QMap in Esprit?
}
} Message: hi all, are there anybody using Bruker Esprit for elemental
} mapping in TEM? Could you please inspire me a bit about what's the
} difference between Mapping, HyperMap and QMap? Which one gives the real
} elemental content and distribution image? What I found is, after
} processing the Hypermaps using QMap, many tiny but shiny spots come out
} which actually don't make sense to me and the material. Thank you!

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Dear Jason,

About the differences between Mapping, HyperMap and QMap, have a look to
the /"Man_QUANTAX_en.pdf/" that came with your Bruker system. There is
there a clear description in a few lines.

More interesting: the "/shiny spots come out which actually don't make
sense to me and the material" are most probably due to the use of the
"Automatic Map Filter/".

This tool is not described in the documentation but I was told it
contains a routine that adapt the filtering procedure/strength to the
signal/noise in the map.

On maps with a fairly high S/N it works well, but on noisy maps it
groups some noise pixels and create pseudo-nanoparticles that do not
exist. Nice images, often meeting our expectancies... but wrong!

This is in particular the case when you didn't accumulate enough counts
in the Hypermap and QMap quantification warns you : "/Quantification:
Very short acquisition time or low count rate. Quantification results
may be inaccurate. Ignore or Cancel procedure/". There is good chances
if you choose "Ignore" that the quantification will be very inaccurate
and your Automatic filter will build pseudo-nanoparticles that disappear
if you choose no filter or another one.

An advice: it's a good practice to always check how looks a spectrum
extracted from the Hypermap over a square area of the same size as the
QMap resolution (1: 1x1=1; 1/2: 2x2=4; 1/4: 4x4=16; 1/8: 8x8=64pxl; you
read the area size on the left of the legend under the hypermap when
"Area unit" is set to pixels). Then you will judge how far can the
Bremsstrahlung subtraction and quantification be accurate.

Enjoy, with regards

Philippe Buffat

--

Prof. Em. Philippe A. Buffat
Ecole Polytechnique Federale de Lausanne (http://cime.epfl.ch
{http://cime.epfl.ch/} )
AGH-UST University of Science and Technology Krakow
(http://www.tem.agh.edu.pl {http://www.tem.agh.edu.pl/} )

{http://cime.epfl.ch} Subimage, ch. des Vioz 14, Chalet l'Amadou
CH-1865 Les Diablerets, Switzerland
mobile +41 79 337 9384
philippe.buffat-at-epfl.ch {mailto:philippe.buffat-at-epfl.ch}

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Hi all,

Thanks for the replies so far! I've had a couple of people asking for more details.

Our instrument is a Hitachi S-4800.
Also regading the issue we could have been more precise, here's the full story:

There are two parameters that influence the probe. On one hand there is a "cond lens setting" which can be altered in the range from 1 to 16 and on the other hand there is a "probe current setting" which can be altered from norm to high. We are wondering which setting influences which lens in what way. Adam wrote that they are both altering C1, just in different ranges. That's sort of the information we were looking for. Further replies are still very welcome.

Regards

Bettina and Sascha

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Ravi;
My advice is forget about auto fill systems. The systems are unreliable. They either fail to turn on, which is a minor problem if your detector has a working bias shut-off (major if not), or worse, they fail to shut off, which empties the liquid nitrogen supply all over your microscope and floor and can cause serious damage. Also, any arrangement that gets them to not touch the microscope and transmit vibration exposes the dewar to introduction of moisture, which induces ice crystals inside the dewar, which degrades performance.

John Mardinly, ASU

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X-from: zaluzec-at-microscopy.com [mailto:zaluzec-at-microscopy.com]
Sent: Monday, November 17, 2014 7:52 PM
To: John Mardinly

X-from: ravithakkar-at-vet.k-state.edu ()

I am inclined to agree with John. I suggest you save the money you would spend on the auto fill system and keep saving to buy an SDD detector to get rid of the lN2 completely (from the XEDS systems). They are not cheap but they are a lot less hassle than Si-Lis

____
John Mansfield PhD Cphys MInstP
*****Note New Business Mailing Address, all other information remains the same*****
Microscopy Society of America - President-Elect
Associate Director
North Campus Electron Microbeam Analysis Laboratory
Building 22, Room G010, University of Michigan
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} On Nov 19, 2014, at 1:51 PM, John.Mardinly-at-asu.edu wrote:
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} Ravi;
} My advice is forget about auto fill systems. The systems are unreliable. They either fail to turn on, which is a minor problem if your detector has a working bias shut-off (major if not), or worse, they fail to shut off, which empties the liquid nitrogen supply all over your microscope and floor and can cause serious damage. Also, any arrangement that gets them to not touch the microscope and transmit vibration exposes the dewar to introduction of moisture, which induces ice crystals inside the dewar, which degrades performance.
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} John Mardinly, ASU
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Contents Retrieved from Microscopy Listserver Archives
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The benefits of going with SDD are much greater than just not having to
deal with the LN2. The systems are so much more sensitive and you get so
many more counts that the time saved is an added bonus. They result in
much more accurate results due to the high count rate, efficiency of the
systems and more sophisticated software.

While at Purdue, I justified replacing a 2-yr old SiLi system with the SDD
to my administration partially on the safety considerations. A picture of
a technician up on a stool pouring LN2 into the dewar and the obvious
hazards involved made a strong argument for getting internal funding for
the replacement.

Debby

Debra Sherman, Founder & CSO
DS imaging LLC
Purdue Technology Center
1281 Win Hentschel Blvd
West Lafayette, IN 47906
E-mail: debby.sherman-at-dsimagingllc.com
www.dsimagingllc.com

On 11/19/14, 9:20 PM, "jfmjfm-at-umich.edu" {jfmjfm-at-umich.edu} wrote:

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10, 37 -- From dsherman-at-purdue.edu Wed Nov 19 20:52:49 2014
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Dear All,
taking the cue from Debby's reply: is there anyone filling the dewar in
a better way than staying on a stool?
I discussed the topic with NMR people here, they use a silicone tube
connected directly to a LN2 tank (correct word?). Does anyone use a
similar system? Does anyone see problems in this method? Maybe
degradation of the material, falling into the XEDS dewar.
Thanks

Davide

~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~~~~~~~
Davide Cristofori

direct phone = 041.234.67.26
e-mail = dcristofori[at]unive[dot]it

Università Ca' Foscari Venezia
Dipartimento di Sc. Molecolari e Nanosistemi
Via Torino, 155b I-30172 Mestre (VE) Italy
~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~~~~~~~

Il 20/11/2014 3.58, dsherman-at-purdue.edu ha scritto:
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} The benefits of going with SDD are much greater than just not having to
} deal with the LN2. The systems are so much more sensitive and you get so
} many more counts that the time saved is an added bonus. They result in
} much more accurate results due to the high count rate, efficiency of the
} systems and more sophisticated software.
}
} While at Purdue, I justified replacing a 2-yr old SiLi system with the SDD
} to my administration partially on the safety considerations. A picture of
} a technician up on a stool pouring LN2 into the dewar and the obvious
} hazards involved made a strong argument for getting internal funding for
} the replacement.
}
} Debby
}
}
} Debra Sherman, Founder & CSO
} DS imaging LLC
} Purdue Technology Center
} 1281 Win Hentschel Blvd
} West Lafayette, IN 47906
} E-mail: debby.sherman-at-dsimagingllc.com
} www.dsimagingllc.com
}
}
} On 11/19/14, 9:20 PM, "jfmjfm-at-umich.edu" {jfmjfm-at-umich.edu} wrote:
}
} }
} }
} }
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} } I am inclined to agree with John. I suggest you save the money you would
} } spend on the auto fill system and keep saving to buy an SDD detector to
} } get rid of the lN2 completely (from the XEDS systems). They are not
} } cheap but they are a lot less hassle than Si-Lis
} }
} } ____
} } John Mansfield PhD Cphys MInstP
} } *****Note New Business Mailing Address, all other information remains the
} } same*****
} } Microscopy Society of America - President-Elect
} } Associate Director
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} }
} } Home address:
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} }
} } } On Nov 19, 2014, at 1:51 PM, John.Mardinly-at-asu.edu wrote:
} } }
} } }
} } }
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} } }
} } } Ravi;
} } } My advice is forget about auto fill systems. The systems are
} } } unreliable. They either fail to turn on, which is a minor problem if
} } } your detector has a working bias shut-off (major if not), or worse, they
} } } fail to shut off, which empties the liquid nitrogen supply all over your
} } } microscope and floor and can cause serious damage. Also, any arrangement
} } } that gets them to not touch the microscope and transmit vibration
} } } exposes the dewar to introduction of moisture, which induces ice
} } } crystals inside the dewar, which degrades performance.
} } }
} } } John Mardinly, ASU
} } }
} } }
} } } -----Original Message-----
} } } X-from: zaluzec-at-microscopy.com [mailto:zaluzec-at-microscopy.com]
} } } Sent: Monday, November 17, 2014 7:52 PM
} } } To: John Mardinly
} } } Subject: [Microscopy] viaWWW:LN2 Filling system
} } }
} } }
} } }
} } }
} } }
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} } } Name: Ravi
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} } } Title-Subject: [Filtered] LN2 Filling system
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} 10, 37 -- From dsherman-at-purdue.edu Wed Nov 19 20:52:49 2014
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6, 22 -- From dcristofori-at-unive.it Thu Nov 20 12:46:55 2014
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Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Years ago when we got our first SEM/EDX system, I quickly learned that I did not like filling the detector using a portable dewar, for all the usual reasons stated by others. So, for 25+ years we filled our SiLi detector using a Swagelok corrugated/braided stainless steel hose. The large LN2 tank was placed adjacent to the SEM so only 4 feet of flex hose was needed to reach the detector. I always made sure I was present when the tanks were changed out to prevent damage to the instrument. Once our delivery folks got the tank to the SEM room, I would tilt/roll the tank into final position. Not hard to do with reasonable strength and a little understanding. The straight hose won't drop right into the detector dewar, so I wired a 90 degree bend into the end of the hose and mounted a sintered diffuser to it to minimize the effects of the initial gassing. To fill, crack the LN2 valve open and chill the hose down, then open it up and fill the detector. The cold hose would condense CO2 out of the air, so gathering and playing with liquid CO2 was a pastime during filling. By listening to the tone of the LN2 whooshing into the tank, it's easy to tell when the detector was getting full.

Since those early days, we've finally seen the light; SDD's are the only way to go!

Rick

Richard A. Ross
Sr. Metallurgist, Materials Engineering
Office: (317) 242-4880 - Fax (317) 242-7638
Richard.Ross-at-AllisonTransmission.com

-----Original Message-----
X-from: dcristofori-at-unive.it [mailto:dcristofori-at-unive.it]
Sent: Thursday, November 20, 2014 2:08 PM
To: Richard A. Ross

Dear All,
taking the cue from Debby's reply: is there anyone filling the dewar in a better way than staying on a stool?
I discussed the topic with NMR people here, they use a silicone tube connected directly to a LN2 tank (correct word?). Does anyone use a similar system? Does anyone see problems in this method? Maybe degradation of the material, falling into the XEDS dewar.
Thanks

Davide

~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~~~~~~~ Davide Cristofori

direct phone = 041.234.67.26
e-mail = dcristofori[at]unive[dot]it

Università Ca' Foscari Venezia
Dipartimento di Sc. Molecolari e Nanosistemi Via Torino, 155b I-30172 Mestre (VE) Italy ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~~~~~~~

Il 20/11/2014 3.58, dsherman-at-purdue.edu ha scritto:
} ----------------------------------------------------------------------
} ------ The Microscopy ListServer -- CoSponsor: The Microscopy Society
} of America To Subscribe/Unsubscribe --
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} The benefits of going with SDD are much greater than just not having
} to deal with the LN2. The systems are so much more sensitive and you
} get so many more counts that the time saved is an added bonus. They
} result in much more accurate results due to the high count rate,
} efficiency of the systems and more sophisticated software.
}
} While at Purdue, I justified replacing a 2-yr old SiLi system with the
} SDD to my administration partially on the safety considerations. A
} picture of a technician up on a stool pouring LN2 into the dewar and
} the obvious hazards involved made a strong argument for getting
} internal funding for the replacement.
}
} Debby
}
}
} Debra Sherman, Founder & CSO
} DS imaging LLC
} Purdue Technology Center
} 1281 Win Hentschel Blvd
} West Lafayette, IN 47906
} E-mail: debby.sherman-at-dsimagingllc.com www.dsimagingllc.com
}
}
} On 11/19/14, 9:20 PM, "jfmjfm-at-umich.edu" {jfmjfm-at-umich.edu} wrote:
}
} }
} }
} }
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} }
} } I am inclined to agree with John. I suggest you save the money you would
} } spend on the auto fill system and keep saving to buy an SDD detector to
} } get rid of the lN2 completely (from the XEDS systems). They are not
} } cheap but they are a lot less hassle than Si-Lis
} }
} } ____
} } John Mansfield PhD Cphys MInstP
} } *****Note New Business Mailing Address, all other information remains the
} } same*****
} } Microscopy Society of America - President-Elect
} } Associate Director
} } North Campus Electron Microbeam Analysis Laboratory
} } Building 22, Room G010, University of Michigan
} } 2800 Plymouth Road, Ann Arbor MI 48109-2800 USA
} } Phone: (734) 936-3352 FAX (734) 763-2282
} } Cell. Phone: (734) 834-3913
} } Email: jfmjfm-at-umich.edu
} } URL: http://emalwww.engin.umich.edu/
} } iMessage: thejfmjfm
} } Skype: thejfmjfm
} }
} } Home address:
} } 4304 Spring Lake Boulevard
} } Ann Arbor MI 48108-9657
} } Phone (734) 994-3096
} }
} } } On Nov 19, 2014, at 1:51 PM, John.Mardinly-at-asu.edu wrote:
} } }
} } }
} } }
} } }
} } }
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} } }
} } } Ravi;
} } } My advice is forget about auto fill systems. The systems are
} } } unreliable. They either fail to turn on, which is a minor problem if
} } } your detector has a working bias shut-off (major if not), or worse, they
} } } fail to shut off, which empties the liquid nitrogen supply all over your
} } } microscope and floor and can cause serious damage. Also, any arrangement
} } } that gets them to not touch the microscope and transmit vibration
} } } exposes the dewar to introduction of moisture, which induces ice
} } } crystals inside the dewar, which degrades performance.
} } }
} } } John Mardinly, ASU
} } }
} } }
} } } -----Original Message-----
} } } X-from: zaluzec-at-microscopy.com [mailto:zaluzec-at-microscopy.com]
} } } Sent: Monday, November 17, 2014 7:52 PM
} } } To: John Mardinly
} } } Subject: [Microscopy] viaWWW:LN2 Filling system
} } }
} } }
} } }
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} } } X-from: ravithakkar-at-vet.k-state.edu ()
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} } } Email: ravithakkar-at-vet.k-state.edu
} } } Name: Ravi
} } }
} } } Title-Subject: [Filtered] LN2 Filling system
} } }
} } } Message: Hi,
} } } I am looking for automated LN2 filling/ transfer system for EDS
} } } installed on scopes. Kindly share some suggestions based on your
} } } experience.
} } }
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} } } 22, 40 -- From John.Mardinly-at-asu.edu Wed Nov 19 12:37:45 2014
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} } } 22, 40 -- Wed, 19 Nov 2014 11:37:43 -0700
} } } 22, 40 -- From: John Mardinly {John.Mardinly-at-asu.edu}
} } } 22, 40 -- To: "zaluzec-at-microscopy.com" {zaluzec-at-Microscopy.com} ,
} } } 22, 40 -- "ravithakkar-at-vet.k-state.edu"
} } } {ravithakkar-at-vet.k-state.edu} ,
} } } 22, 40 -- "Microscopy-at-Microscopy.com"
} } } {Microscopy-at-Microscopy.com}
} } } 22, 40 -- Subject: RE: [Microscopy] viaWWW:LN2 Filling system
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} } 6, 37 -- From jfmjfm-at-umich.edu Wed Nov 19 20:18:01 2014
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} 10, 37 -- From dsherman-at-purdue.edu Wed Nov 19 20:52:49 2014
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} 10, 37 -- Nov 2014 21:52:47 -0500
} 10, 37 -- From: "Sherman, Debra" {dsherman-at-purdue.edu}
} 10, 37 -- To: "jfmjfm-at-umich.edu" {jfmjfm-at-umich.edu} ,
} 10, 37 -- "ravithakkar-at-vet.k-state.edu"
} 10, 37 -- {ravithakkar-at-vet.k-state.edu} ,
} 10, 37 -- "message to: MSA list"
} 10, 37 -- {microscopy-at-microscopy.com}
} 10, 37 -- Subject: Re: [Microscopy] viaWWW:LN2 Filling system
} 10, 37 -- Thread-Topic: [Microscopy] viaWWW:LN2 Filling system
} 10, 37 -- Thread-Index: AQHQBGiSmB7SELn4J0WEjg/xkfPHXpxo0UuA
} 10, 37 -- Date: Thu, 20 Nov 2014 02:52:46 +0000
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6, 22 -- From dcristofori-at-unive.it Thu Nov 20 12:46:55 2014
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16, 36 -- From richard.ross-at-allisontransmission.com Thu Nov 20 14:09:25 2014
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16, 36 -- From: "Richard A. Ross" {richard.ross-at-allisontransmission.com}
16, 36 -- To: "Microscopy-at-Microscopy.com" {Microscopy-at-Microscopy.com}
16, 36 -- Subject: RE: [Microscopy] viaWWW:LN2 Filling system
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Title-Subject: [Filtered] EDS-DT-Problem

Message: hello

we have a problem with EDS, Si(Li) detectors (JEOL-JSM6360LV) with super Ultra Thin Window SUTW
the Dead times DT, is always 100% don't want to decrease to.
as trying to optimized by adjusting the beam current (probe current or spot size), and/or the
process time
are there any solutions for these problems.

Best regards

Naitbouda Abdelyamine

SEM Lab service Engineer at advanced technologies Development Center -CDTA
Algers ALGERIA

www.cdta.dz
anaitbouda-at-cdta.dz

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You did not mention which EDS detector or system you have. That may have some bearing in the trouble-shooting. Is this a new system to you or did the problem just occur? What is the background history?

What happens if you try to collect a spectrum with the beam turned completely off or blanked? If you still have excessive counts and deadtime, you could have a problem somewhere in the EDS system.

Are you sure there is no other source of light hitting the detector? Chamber cameras are nice to have, but they cannot be on while trying to collect x-rays. The myriad IR photons will flood most EDS detectors.

Is there a way of monitoring the count rate and/or reset pulse on your system. On a very old EDS system with external amplifiers, it was possible to adjust the discriminators by watching lights on the pulse processor. You wanted to have a very low level of counts with the beam off. If the discriminators were set too low, excessive noise slipped through and increased the baseline count rate. New systems hide those adjustments in areas typically reserved for engineers, but adjustments can still be made. You may have to call for expert help.

Warren Straszheim

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Title-Subject: [Filtered] EDS-DT-Problem

Message: hello

we have a problem with EDS, Si(Li) detectors (JEOL-JSM6360LV) with super Ultra Thin Window SUTW the Dead times DT, is always 100% don't want to decrease to.
as trying to optimized by adjusting the beam current (probe current or spot size), and/or the process time are there any solutions for these problems.

Best regards

Naitbouda Abdelyamine

SEM Lab service Engineer at advanced technologies Development Center -CDTA Algers ALGERIA

www.cdta.dz
anaitbouda-at-cdta.dz

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Hello Davide,
The problem with the silicon tube method is, that you first have to get the tube properly cooled down until no more gas but really liquid becomes transported. To control that initial phase safely immersing the tube directly to the EXDS dewar might be even more difficult than externally filling first your small dewar, away from the microscopy at a safe space, and then walk with the small dewar to the XEDS dewar via a stable stairway. I wouldn't be surprised if the initial strong gas flow directly going into the XEDS dewar would even damage something in the XEDS dewar on the long term.
I would not spend too much time on thinking about an automatic filling system, but just think about how to replace your stool by a proper, stable, fixed-in-place stairway.

Best greetings from the EM-Labs of CIC biomaGUNE,
Marco Möller

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Marco Möller
Platform Manager - Electron Microscopy

CIC biomaGUNE
Parque Tecnológico de San Sebastián
Paseo Miramón, 182. Ed. Empresarial C
20009 San Sebastián (Guipúzcoa)
SPAIN

Tel: +34 943 00 53 25
mmoller-at-cicbiomagune.es

-----Original Message-----
X-from: dcristofori-at-unive.it [mailto:dcristofori-at-unive.it]
Sent: Thursday, November 20, 2014 8:04 PM
To: Marco Moller

Dear All,
taking the cue from Debby's reply: is there anyone filling the dewar in a better way than staying on a stool?
I discussed the topic with NMR people here, they use a silicone tube connected directly to a LN2 tank (correct word?). Does anyone use a similar system? Does anyone see problems in this method? Maybe degradation of the material, falling into the XEDS dewar.
Thanks

Davide

~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~~~~~~~ Davide Cristofori

direct phone = 041.234.67.26
e-mail = dcristofori[at]unive[dot]it

Università Ca' Foscari Venezia
Dipartimento di Sc. Molecolari e Nanosistemi Via Torino, 155b I-30172 Mestre (VE) Italy ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~~~~~~~

Il 20/11/2014 3.58, dsherman-at-purdue.edu ha scritto:
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} The benefits of going with SDD are much greater than just not having
} to deal with the LN2. The systems are so much more sensitive and you
} get so many more counts that the time saved is an added bonus. They
} result in much more accurate results due to the high count rate,
} efficiency of the systems and more sophisticated software.
}
} While at Purdue, I justified replacing a 2-yr old SiLi system with the
} SDD to my administration partially on the safety considerations. A
} picture of a technician up on a stool pouring LN2 into the dewar and
} the obvious hazards involved made a strong argument for getting
} internal funding for the replacement.
}
} Debby
}
}
} Debra Sherman, Founder & CSO
} DS imaging LLC
} Purdue Technology Center
} 1281 Win Hentschel Blvd
} West Lafayette, IN 47906
} E-mail: debby.sherman-at-dsimagingllc.com www.dsimagingllc.com
}
}
} On 11/19/14, 9:20 PM, "jfmjfm-at-umich.edu" {jfmjfm-at-umich.edu} wrote:
}
} }
} }
} }
} } --------------------------------------------------------------------------
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} } I am inclined to agree with John. I suggest you save the money you would
} } spend on the auto fill system and keep saving to buy an SDD detector to
} } get rid of the lN2 completely (from the XEDS systems). They are not
} } cheap but they are a lot less hassle than Si-Lis
} }
} } ____
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} } *****Note New Business Mailing Address, all other information remains the
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} } Microscopy Society of America - President-Elect
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} } } On Nov 19, 2014, at 1:51 PM, John.Mardinly-at-asu.edu wrote:
} } }
} } }
} } }
} } }
} } }
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} } } Ravi;
} } } My advice is forget about auto fill systems. The systems are
} } } unreliable. They either fail to turn on, which is a minor problem if
} } } your detector has a working bias shut-off (major if not), or worse, they
} } } fail to shut off, which empties the liquid nitrogen supply all over your
} } } microscope and floor and can cause serious damage. Also, any arrangement
} } } that gets them to not touch the microscope and transmit vibration
} } } exposes the dewar to introduction of moisture, which induces ice
} } } crystals inside the dewar, which degrades performance.
} } }
} } } John Mardinly, ASU
} } }
} } }
} } } -----Original Message-----
} } } X-from: zaluzec-at-microscopy.com [mailto:zaluzec-at-microscopy.com]
} } } Sent: Monday, November 17, 2014 7:52 PM
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} } } Subject: [Microscopy] viaWWW:LN2 Filling system
} } }
} } }
} } }
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} } } Name: Ravi
} } }
} } } Title-Subject: [Filtered] LN2 Filling system
} } }
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Contents Retrieved from Microscopy Listserver Archives
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Hello All,

A reminder that the University of Florida has an EM Technician position
posted. The posting will close midnight November 26th, so pass this onto
technicians or recent graduates from any of the microscopy colleges.

Karen

On 11/18/14, 6:44 AM, "vau-at-ufl.edu" {vau-at-ufl.edu} wrote:

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8, 38 -- From vau-at-ufl.edu Fri Nov 21 08:35:16 2014
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8, 38 -- From: "Kelley,Karen L" {vau-at-ufl.edu}
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I am stitching multiple digital images of paraffin sections photographed with either a 4x or 10x objective. A typical composite is 10-40 overlapping sections.

Using my standard MagnaFire digital camera, I have used FIJI software to implement the background smoothing protocol outlined by George McNamara (Color Balancing Histology Images for Presentations and Publication. The Journal of Histotechnology 28(2):81-88 available at http://home.earthlink.net/~geomcnamara/McNamara2005JoH28n2pp81-88.pdf). Essentially you subtract a "black" image from all the images (to remove hot pixels) and then divide by a background white image (no tissue, blank spot on the slide) and multiply the resulting image by some factor such as 220. This works okay but is somewhat tedious.

The method I really think does the best job employs Photoshop. I subtract the black image from every image just like in George's protocol and then use the Photoshop "Apply image - divide" using the background white image to divide the tissue image. It gives a great clean bright background. But this Photoshop function must be multiplying the divided image by something (and possibly doing other things) and, despite extensive online searching, I can find no documentation on what it is doing. I wouldn't try to publish an image where I have done an image processing step that I don't fully understand. Naturally I will describe what I have done in the Methods section - I have no worries on the ethical basis of what I am doing since I am going to explicitly state what I have done. But step one in that explanation is to understand is for me to understand what Photoshop does in this function. Does anyone know exactly what this function does?

Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
phillipst-at-missouri.edu

http://biology.missouri.edu/people/?person=88
http://www.biotech.missouri.edu/mcc/

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7, 33 -- From PhillipsT-at-missouri.edu Fri Nov 21 09:37:30 2014
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7, 33 -- From: "Phillips, Thomas E." {PhillipsT-at-missouri.edu}
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7, 33 -- CC: George McNamara {geomcnamara-at-EARTHLINK.NET} ,
7, 33 -- jerry sedgewick
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7, 33 -- Subject: Using Photoshop for creating an even background field for LM montage
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The Apply Image - Divide is a pixel by pixel divide from what I've
tested. It's the same as using Divide in the layer mode functions
(http://en.wikipedia.org/wiki/Blend_modes).

Jerry

On 11/21/2014 9:49 AM, PhillipsT-at-missouri.edu wrote:
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} I am stitching multiple digital images of paraffin sections photographed with either a 4x or 10x objective. A typical composite is 10-40 overlapping sections.
}
} Using my standard MagnaFire digital camera, I have used FIJI software to implement the background smoothing protocol outlined by George McNamara (Color Balancing Histology Images for Presentations and Publication. The Journal of Histotechnology 28(2):81-88 available at http://home.earthlink.net/~geomcnamara/McNamara2005JoH28n2pp81-88.pdf). Essentially you subtract a "black" image from all the images (to remove hot pixels) and then divide by a background white image (no tissue, blank spot on the slide) and multiply the resulting image by some factor such as 220. This works okay but is somewhat tedious.
}
} The method I really think does the best job employs Photoshop. I subtract the black image from every image just like in George's protocol and then use the Photoshop "Apply image - divide" using the background white image to divide the tissue image. It gives a great clean bright background. But this Photoshop function must be multiplying the divided image by something (and possibly doing other things) and, despite extensive online searching, I can find no documentation on what it is doing. I wouldn't try to publish an image where I have done an image processing step that I don't fully understand. Naturally I will describe what I have done in the Methods section - I have no worries on the ethical basis of what I am doing since I am going to explicitly state what I have done. But step one in that explanation is to understand is for me to understand what Photoshop does in this function. Does anyone know exactly what this function does?
}
}
} Thomas E. Phillips, Ph.D
} Professor of Biological Sciences
} Director, Molecular Cytology Core
} 2 Tucker Hall
} University of Missouri
} Columbia, MO 65211-7400
} 573-882-4712 (office)
} 573-882-0123 (fax)
} phillipst-at-missouri.edu
}
} http://biology.missouri.edu/people/?person=88
} http://www.biotech.missouri.edu/mcc/
}
}
}
} ==============================Original Headers==============================
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} 7, 33 -- jerry sedgewick
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} 7, 33 -- Subject: Using Photoshop for creating an even background field for LM montage
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Once you have divided by your illumination image (the one with no sample), your result is a ratio of your sample to a constant illumination, giving the fraction of light that your sample absorbs. Rescaling at that point is somewhat arbitrary, but the most common choices are to rescale by the scalar mean of the illumination image or possibly the scalar max. This is what is done in many software packages - Image Pro (our product, commercial disclaimer here) does this with the illumination mean.

Image' = [ (Image - BlackIm) / Illumination ] * mean(Illumination)

or

Image' = [ (Image - BlackIm) / Illumination ] * max(Illumination)

Either rescales the processed image into the same rough integer range as the original. Using the max( ) gives brighter results, with less precision loss due to rounding with a brighter image overall, but risks saturation and clipping. Rescaling by mean( ) is generally a safer choice. Again, the final result is somewhat arbitrary, but that doesn't really matter - even with _perfectly_ flat illumination that didn't need correction (something I've never managed with a microscope) you can get different intensity final images just by changing the exposure time. What you gain with correction is a shift-invariant response of your sample.

You can crosscheck the Photoshop method by doing these two alternatives by hand. If your step by step results match Photoshop, you've identified what scalar they use to bring the image back into integer range.

Side note: you can do the same with fluorescent images, imaging a stained glass test slide (Chroma makes these) or a volume of dye in solution (gasket on a slide filled to slight excess, drop a cover slip on it, image mid-volume) to obtain your illumination image - thus characterizing spatial flatness for your microscope.

Kevin Ryan
Media Cybernetics, Inc.

-----Original Message-----
X-from: PhillipsT-at-missouri.edu [mailto:PhillipsT-at-missouri.edu]
Sent: Friday, November 21, 2014 10:50 AM
To: Kevin Ryan

I am stitching multiple digital images of paraffin sections photographed with either a 4x or 10x objective. A typical composite is 10-40 overlapping sections.

Using my standard MagnaFire digital camera, I have used FIJI software to implement the background smoothing protocol outlined by George McNamara (Color Balancing Histology Images for Presentations and Publication. The Journal of Histotechnology 28(2):81-88 available at http://home.earthlink.net/~geomcnamara/McNamara2005JoH28n2pp81-88.pdf). Essentially you subtract a "black" image from all the images (to remove hot pixels) and then divide by a background white image (no tissue, blank spot on the slide) and multiply the resulting image by some factor such as 220. This works okay but is somewhat tedious.

The method I really think does the best job employs Photoshop. I subtract the black image from every image just like in George's protocol and then use the Photoshop "Apply image - divide" using the background white image to divide the tissue image. It gives a great clean bright background. But this Photoshop function must be multiplying the divided image by something (and possibly doing other things) and, despite extensive online searching, I can find no documentation on what it is doing. I wouldn't try to publish an image where I have done an image processing step that I don't fully understand. Naturally I will describe what I have done in the Methods section - I have no worries on the ethical basis of what I am doing since I am going to explicitly state what I have done. But step one in that explanation is to understand is for me to understand what Photoshop does in this function. Does anyone know exactly what this function does?

Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
phillipst-at-missouri.edu

http://biology.missouri.edu/people/?person=88
http://www.biotech.missouri.edu/mcc/

==============================Original Headers==============================
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7, 33 -- jerry sedgewick
7, 33 -- {jerrysedgewick-at-GMAIL.COM}
7, 33 -- Subject: Using Photoshop for creating an even background field for LM montage 7, 33 -- Thread-Topic: Using Photoshop for creating an even background field for LM 7, 33 -- montage 7, 33 -- Thread-Index: AdAFGUSUQK8c4SD4QfaTOZjdjjij2gAhbdbwAABzqfA=
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22, 28 -- From KRyan-at-mediacy.com Fri Nov 21 11:56:51 2014
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22, 28 -- To: "Microscopy-at-microscopy.com" {Microscopy-at-microscopy.com}
22, 28 -- Subject: RE: [Microscopy] Using Photoshop for creating an even background
22, 28 -- field for LM montage
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Thanks to Jerry and Kevin and everyone else who replied on my question. I don't know why I didn't think of doing the obvious thing and making a couple of homogeneous grey scale images and then dividing by some homogeneous brighter "grey" scale image and looking at the resulting pixels. To confirm Jerry's statement, I made a Photoshop image where I created some large bands of 120, 150, 175, 200 grey pixel values and then divided by various brighter images such as one whose pixels were all 220. The resulting bands were indeed the result of x/220 * 255. If x } the denominator (i.e., brighter than the background white image), it causes saturation. Tom

Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
phillipst-at-missouri.edu

http://biology.missouri.edu/people/?person=88
http://www.biotech.missouri.edu/mcc/

-----Original Message-----
X-from: KRyan-at-mediacy.com [mailto:KRyan-at-mediacy.com]
Sent: Friday, November 21, 2014 11:58 AM
To: Phillips, Thomas E.

Once you have divided by your illumination image (the one with no sample), your result is a ratio of your sample to a constant illumination, giving the fraction of light that your sample absorbs. Rescaling at that point is somewhat arbitrary, but the most common choices are to rescale by the scalar mean of the illumination image or possibly the scalar max. This is what is done in many software packages - Image Pro (our product, commercial disclaimer here) does this with the illumination mean.

Image' = [ (Image - BlackIm) / Illumination ] * mean(Illumination)

or

Image' = [ (Image - BlackIm) / Illumination ] * max(Illumination)

Either rescales the processed image into the same rough integer range as the original. Using the max( ) gives brighter results, with less precision loss due to rounding with a brighter image overall, but risks saturation and clipping. Rescaling by mean( ) is generally a safer choice. Again, the final result is somewhat arbitrary, but that doesn't really matter - even with _perfectly_ flat illumination that didn't need correction (something I've never managed with a microscope) you can get different intensity final images just by changing the exposure time. What you gain with correction is a shift-invariant response of your sample.

You can crosscheck the Photoshop method by doing these two alternatives by hand. If your step by step results match Photoshop, you've identified what scalar they use to bring the image back into integer range.

Side note: you can do the same with fluorescent images, imaging a stained glass test slide (Chroma makes these) or a volume of dye in solution (gasket on a slide filled to slight excess, drop a cover slip on it, image mid-volume) to obtain your illumination image - thus characterizing spatial flatness for your microscope.

Kevin Ryan
Media Cybernetics, Inc.

-----Original Message-----
X-from: PhillipsT-at-missouri.edu [mailto:PhillipsT-at-missouri.edu]
Sent: Friday, November 21, 2014 10:50 AM
To: Kevin Ryan

I am stitching multiple digital images of paraffin sections photographed with either a 4x or 10x objective. A typical composite is 10-40 overlapping sections.

Using my standard MagnaFire digital camera, I have used FIJI software to implement the background smoothing protocol outlined by George McNamara (Color Balancing Histology Images for Presentations and Publication. The Journal of Histotechnology 28(2):81-88 available at http://home.earthlink.net/~geomcnamara/McNamara2005JoH28n2pp81-88.pdf). Essentially you subtract a "black" image from all the images (to remove hot pixels) and then divide by a background white image (no tissue, blank spot on the slide) and multiply the resulting image by some factor such as 220. This works okay but is somewhat tedious.

The method I really think does the best job employs Photoshop. I subtract the black image from every image just like in George's protocol and then use the Photoshop "Apply image - divide" using the background white image to divide the tissue image. It gives a great clean bright background. But this Photoshop function must be multiplying the divided image by something (and possibly doing other things) and, despite extensive online searching, I can find no documentation on what it is doing. I wouldn't try to publish an image where I have done an image processing step that I don't fully understand. Naturally I will describe what I have done in the Methods section - I have no worries on the ethical basis of what I am doing since I am going to explicitly state what I have done. But step one in that explanation is to understand is for me to understand what Photoshop does in this function. Does anyone know exactly what this function does?

Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
phillipst-at-missouri.edu

http://biology.missouri.edu/people/?person=88
http://www.biotech.missouri.edu/mcc/

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Oh my. What you have described Photoshop doing is rescaling to the max integer value possible for the data type. That's _probably_ OK for transmitted light, where you expect any sample to absorb and be darker than the flat field image, and as long as you ensure that your flat field image is taken with the same exposure time as your sample.

But rescaling to the range of the image date type is wholly inappropriate for fluorescence, as the staining in your sample may or _may not_ be brighter than your flat field reference. If you apply that Photoshop function to flat fielding fluorescence you'll have saturation/clipping of anything brighter than the reference. That's why IMO rescaling by the flat field mean is more appropriate - areas illuminated more than average get dimmed, areas illuminated less get amplified, all moving towards the sample image mid-range, and (on average) the overall intensity of the sample doesn't change under correction.

Kevin Ryan
Media Cybernetics, Inc.

-----Original Message-----
X-from: Phillips, Thomas E. [mailto:PhillipsT-at-missouri.edu]
Sent: Friday, November 21, 2014 4:02 PM
To: 'microscopy-at-microscopy.com'
Cc: Kevin Ryan; jerry sedgewick

Once you have divided by your illumination image (the one with no sample), your result is a ratio of your sample to a constant illumination, giving the fraction of light that your sample absorbs. Rescaling at that point is somewhat arbitrary, but the most common choices are to rescale by the scalar mean of the illumination image or possibly the scalar max. This is what is done in many software packages - Image Pro (our product, commercial disclaimer here) does this with the illumination mean.

Image' = [ (Image - BlackIm) / Illumination ] * mean(Illumination)

or

Image' = [ (Image - BlackIm) / Illumination ] * max(Illumination)

Either rescales the processed image into the same rough integer range as the original. Using the max( ) gives brighter results, with less precision loss due to rounding with a brighter image overall, but risks saturation and clipping. Rescaling by mean( ) is generally a safer choice. Again, the final result is somewhat arbitrary, but that doesn't really matter - even with _perfectly_ flat illumination that didn't need correction (something I've never managed with a microscope) you can get different intensity final images just by changing the exposure time. What you gain with correction is a shift-invariant response of your sample.

You can crosscheck the Photoshop method by doing these two alternatives by hand. If your step by step results match Photoshop, you've identified what scalar they use to bring the image back into integer range.

Side note: you can do the same with fluorescent images, imaging a stained glass test slide (Chroma makes these) or a volume of dye in solution (gasket on a slide filled to slight excess, drop a cover slip on it, image mid-volume) to obtain your illumination image - thus characterizing spatial flatness for your microscope.

Kevin Ryan
Media Cybernetics, Inc.

-----Original Message-----
X-from: PhillipsT-at-missouri.edu [mailto:PhillipsT-at-missouri.edu]
Sent: Friday, November 21, 2014 10:50 AM
To: Kevin Ryan

I am stitching multiple digital images of paraffin sections photographed with either a 4x or 10x objective. A typical composite is 10-40 overlapping sections.

Using my standard MagnaFire digital camera, I have used FIJI software to implement the background smoothing protocol outlined by George McNamara (Color Balancing Histology Images for Presentations and Publication. The Journal of Histotechnology 28(2):81-88 available at http://home.earthlink.net/~geomcnamara/McNamara2005JoH28n2pp81-88.pdf). Essentially you subtract a "black" image from all the images (to remove hot pixels) and then divide by a background white image (no tissue, blank spot on the slide) and multiply the resulting image by some factor such as 220. This works okay but is somewhat tedious.

The method I really think does the best job employs Photoshop. I subtract the black image from every image just like in George's protocol and then use the Photoshop "Apply image - divide" using the background white image to divide the tissue image. It gives a great clean bright background. But this Photoshop function must be multiplying the divided image by something (and possibly doing other things) and, despite extensive online searching, I can find no documentation on what it is doing. I wouldn't try to publish an image where I have done an image processing step that I don't fully understand. Naturally I will describe what I have done in the Methods section - I have no worries on the ethical basis of what I am doing since I am going to explicitly state what I have done. But step one in that explanation is to understand is for me to understand what Photoshop does in this function. Does anyone know exactly what this function does?

Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
phillipst-at-missouri.edu

http://biology.missouri.edu/people/?person=88
http://www.biotech.missouri.edu/mcc/

==============================Original Headers==============================
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I am in full agreement that this would be wrong for fluorescence images since saturation can lead to an artifactual representation of the real image. My sections are all H&E stained paraffin sections and I am simply trying to illustrate the overall morphology of a large field of view. All the images, including the background image, were taken with identical exposures. Saturation of an RGB image of a paraffin section might risk losing detail but not lead to the type of misleading conclusion one would get while working with fluorescence images. One might avoid saturation by rescaling with a mean value but doesn't this lead to a compression since you are using less than the full 256 pixel range?

Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
phillipst-at-missouri.edu

http://biology.missouri.edu/people/?person=88
http://www.biotech.missouri.edu/mcc/

-----Original Message-----
X-from: Kevin Ryan [mailto:KRyan-at-mediacy.com]
Sent: Friday, November 21, 2014 3:25 PM
To: 'microscopy-at-microscopy.com'
Cc: Phillips, Thomas E.

Once you have divided by your illumination image (the one with no sample), your result is a ratio of your sample to a constant illumination, giving the fraction of light that your sample absorbs. Rescaling at that point is somewhat arbitrary, but the most common choices are to rescale by the scalar mean of the illumination image or possibly the scalar max. This is what is done in many software packages - Image Pro (our product, commercial disclaimer here) does this with the illumination mean.

Image' = [ (Image - BlackIm) / Illumination ] * mean(Illumination)

or

Image' = [ (Image - BlackIm) / Illumination ] * max(Illumination)

Either rescales the processed image into the same rough integer range as the original. Using the max( ) gives brighter results, with less precision loss due to rounding with a brighter image overall, but risks saturation and clipping. Rescaling by mean( ) is generally a safer choice. Again, the final result is somewhat arbitrary, but that doesn't really matter - even with _perfectly_ flat illumination that didn't need correction (something I've never managed with a microscope) you can get different intensity final images just by changing the exposure time. What you gain with correction is a shift-invariant response of your sample.

You can crosscheck the Photoshop method by doing these two alternatives by hand. If your step by step results match Photoshop, you've identified what scalar they use to bring the image back into integer range.

Side note: you can do the same with fluorescent images, imaging a stained glass test slide (Chroma makes these) or a volume of dye in solution (gasket on a slide filled to slight excess, drop a cover slip on it, image mid-volume) to obtain your illumination image - thus characterizing spatial flatness for your microscope.

Kevin Ryan
Media Cybernetics, Inc.

-----Original Message-----
X-from: PhillipsT-at-missouri.edu [mailto:PhillipsT-at-missouri.edu]
Sent: Friday, November 21, 2014 10:50 AM
To: Kevin Ryan

I am stitching multiple digital images of paraffin sections photographed with either a 4x or 10x objective. A typical composite is 10-40 overlapping sections.

Using my standard MagnaFire digital camera, I have used FIJI software to implement the background smoothing protocol outlined by George McNamara (Color Balancing Histology Images for Presentations and Publication. The Journal of Histotechnology 28(2):81-88 available at http://home.earthlink.net/~geomcnamara/McNamara2005JoH28n2pp81-88.pdf). Essentially you subtract a "black" image from all the images (to remove hot pixels) and then divide by a background white image (no tissue, blank spot on the slide) and multiply the resulting image by some factor such as 220. This works okay but is somewhat tedious.

The method I really think does the best job employs Photoshop. I subtract the black image from every image just like in George's protocol and then use the Photoshop "Apply image - divide" using the background white image to divide the tissue image. It gives a great clean bright background. But this Photoshop function must be multiplying the divided image by something (and possibly doing other things) and, despite extensive online searching, I can find no documentation on what it is doing. I wouldn't try to publish an image where I have done an image processing step that I don't fully understand. Naturally I will describe what I have done in the Methods section - I have no worries on the ethical basis of what I am doing since I am going to explicitly state what I have done. But step one in that explanation is to understand is for me to understand what Photoshop does in this function. Does anyone know exactly what this function does?

Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
phillipst-at-missouri.edu

http://biology.missouri.edu/people/?person=88
http://www.biotech.missouri.edu/mcc/

==============================Original Headers==============================
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Rescaling by the mean will show histogram compression where the illumination is higher than average, and histogram expansion where it is lower than average. Scaling up gives only histogram expansion.

But you're going to get (+/-) 1 rounding losses for any scaling whether up or down (rounding direction being pretty much unbiased over the full intensity range and overall image) - I consider the absolute _loss_ of data with saturation far worse than an integer rounding. And that's true even in RGB transmitted microscopy, as you risk losing dim edges of your objects to saturation, shifted automatic thresholds, and undercounted areas.

Quite frankly, if my results (or perceptions) are conditional on a (+/-) 1 intensity value or (+/-) 1 pixel count, they are not strong results.

Kevin Ryan
Media Cybernetics, Inc.

-----Original Message-----
X-from: Phillips, Thomas E. [mailto:PhillipsT-at-missouri.edu]
Sent: Friday, November 21, 2014 5:02 PM
To: Kevin Ryan; 'microscopy-at-microscopy.com'

Once you have divided by your illumination image (the one with no sample), your result is a ratio of your sample to a constant illumination, giving the fraction of light that your sample absorbs. Rescaling at that point is somewhat arbitrary, but the most common choices are to rescale by the scalar mean of the illumination image or possibly the scalar max. This is what is done in many software packages - Image Pro (our product, commercial disclaimer here) does this with the illumination mean.

Image' = [ (Image - BlackIm) / Illumination ] * mean(Illumination)

or

Image' = [ (Image - BlackIm) / Illumination ] * max(Illumination)

Either rescales the processed image into the same rough integer range as the original. Using the max( ) gives brighter results, with less precision loss due to rounding with a brighter image overall, but risks saturation and clipping. Rescaling by mean( ) is generally a safer choice. Again, the final result is somewhat arbitrary, but that doesn't really matter - even with _perfectly_ flat illumination that didn't need correction (something I've never managed with a microscope) you can get different intensity final images just by changing the exposure time. What you gain with correction is a shift-invariant response of your sample.

You can crosscheck the Photoshop method by doing these two alternatives by hand. If your step by step results match Photoshop, you've identified what scalar they use to bring the image back into integer range.

Side note: you can do the same with fluorescent images, imaging a stained glass test slide (Chroma makes these) or a volume of dye in solution (gasket on a slide filled to slight excess, drop a cover slip on it, image mid-volume) to obtain your illumination image - thus characterizing spatial flatness for your microscope.

Kevin Ryan
Media Cybernetics, Inc.

-----Original Message-----
X-from: PhillipsT-at-missouri.edu [mailto:PhillipsT-at-missouri.edu]
Sent: Friday, November 21, 2014 10:50 AM
To: Kevin Ryan

I am stitching multiple digital images of paraffin sections photographed with either a 4x or 10x objective. A typical composite is 10-40 overlapping sections.

Using my standard MagnaFire digital camera, I have used FIJI software to implement the background smoothing protocol outlined by George McNamara (Color Balancing Histology Images for Presentations and Publication. The Journal of Histotechnology 28(2):81-88 available at http://home.earthlink.net/~geomcnamara/McNamara2005JoH28n2pp81-88.pdf). Essentially you subtract a "black" image from all the images (to remove hot pixels) and then divide by a background white image (no tissue, blank spot on the slide) and multiply the resulting image by some factor such as 220. This works okay but is somewhat tedious.

The method I really think does the best job employs Photoshop. I subtract the black image from every image just like in George's protocol and then use the Photoshop "Apply image - divide" using the background white image to divide the tissue image. It gives a great clean bright background. But this Photoshop function must be multiplying the divided image by something (and possibly doing other things) and, despite extensive online searching, I can find no documentation on what it is doing. I wouldn't try to publish an image where I have done an image processing step that I don't fully understand. Naturally I will describe what I have done in the Methods section - I have no worries on the ethical basis of what I am doing since I am going to explicitly state what I have done. But step one in that explanation is to understand is for me to understand what Photoshop does in this function. Does anyone know exactly what this function does?

Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
phillipst-at-missouri.edu

http://biology.missouri.edu/people/?person=88
http://www.biotech.missouri.edu/mcc/

==============================Original Headers==============================
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7, 33 -- Subject: Using Photoshop for creating an even background field for LM montage 7, 33 -- Thread-Topic: Using Photoshop for creating an even background field for LM 7, 33 -- montage 7, 33 -- Thread-Index: AdAFGUSUQK8c4SD4QfaTOZjdjjij2gAhbdbwAABzqfA=
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I am convinced. I guess it is a good thing I didn't think of the obvious use of test images to figure out how Photoshop worked because then I would have just proceeded with that strategy and never started this thread. I will give it a try with the mean value. I appreciate you taking the time to explain things so clearly. Tom

Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
phillipst-at-missouri.edu

http://biology.missouri.edu/people/?person=88
http://www.biotech.missouri.edu/mcc/

-----Original Message-----
X-from: Kevin Ryan [mailto:KRyan-at-mediacy.com]
Sent: Friday, November 21, 2014 4:28 PM
To: Phillips, Thomas E.; 'microscopy-at-microscopy.com'

Once you have divided by your illumination image (the one with no sample), your result is a ratio of your sample to a constant illumination, giving the fraction of light that your sample absorbs. Rescaling at that point is somewhat arbitrary, but the most common choices are to rescale by the scalar mean of the illumination image or possibly the scalar max. This is what is done in many software packages - Image Pro (our product, commercial disclaimer here) does this with the illumination mean.

Image' = [ (Image - BlackIm) / Illumination ] * mean(Illumination)

or

Image' = [ (Image - BlackIm) / Illumination ] * max(Illumination)

Either rescales the processed image into the same rough integer range as the original. Using the max( ) gives brighter results, with less precision loss due to rounding with a brighter image overall, but risks saturation and clipping. Rescaling by mean( ) is generally a safer choice. Again, the final result is somewhat arbitrary, but that doesn't really matter - even with _perfectly_ flat illumination that didn't need correction (something I've never managed with a microscope) you can get different intensity final images just by changing the exposure time. What you gain with correction is a shift-invariant response of your sample.

You can crosscheck the Photoshop method by doing these two alternatives by hand. If your step by step results match Photoshop, you've identified what scalar they use to bring the image back into integer range.

Side note: you can do the same with fluorescent images, imaging a stained glass test slide (Chroma makes these) or a volume of dye in solution (gasket on a slide filled to slight excess, drop a cover slip on it, image mid-volume) to obtain your illumination image - thus characterizing spatial flatness for your microscope.

Kevin Ryan
Media Cybernetics, Inc.

-----Original Message-----
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Sent: Friday, November 21, 2014 10:50 AM
To: Kevin Ryan

I am stitching multiple digital images of paraffin sections photographed with either a 4x or 10x objective. A typical composite is 10-40 overlapping sections.

Using my standard MagnaFire digital camera, I have used FIJI software to implement the background smoothing protocol outlined by George McNamara (Color Balancing Histology Images for Presentations and Publication. The Journal of Histotechnology 28(2):81-88 available at http://home.earthlink.net/~geomcnamara/McNamara2005JoH28n2pp81-88.pdf). Essentially you subtract a "black" image from all the images (to remove hot pixels) and then divide by a background white image (no tissue, blank spot on the slide) and multiply the resulting image by some factor such as 220. This works okay but is somewhat tedious.

The method I really think does the best job employs Photoshop. I subtract the black image from every image just like in George's protocol and then use the Photoshop "Apply image - divide" using the background white image to divide the tissue image. It gives a great clean bright background. But this Photoshop function must be multiplying the divided image by something (and possibly doing other things) and, despite extensive online searching, I can find no documentation on what it is doing. I wouldn't try to publish an image where I have done an image processing step that I don't fully understand. Naturally I will describe what I have done in the Methods section - I have no worries on the ethical basis of what I am doing since I am going to explicitly state what I have done. But step one in that explanation is to understand is for me to understand what Photoshop does in this function. Does anyone know exactly what this function does?

Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
phillipst-at-missouri.edu

http://biology.missouri.edu/people/?person=88
http://www.biotech.missouri.edu/mcc/

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Name: Ahmad Ashkhaibi

Organization: Al-Balqa University

Title-Subject: [Filtered] Venting TECNAI Gun

Message: Hello everybody..

I operate a TECNAI T12 SPIRIT transmission electron microscope. In order to replace the filament,
you have to vent the electron gun (thermoionic), according to the FEI operating manual, before
lifting up the gun cover.

The problem is, when I vent the gun it doesn't vent completely to the atmospheric pressure (log 99),
it keeps holding some vacuum (log 95) that prevents me from lifting up the gun cover to change the
filament.

Has anybody ever experienced anything like that?

Regards,

Ahmad Ashkhaibi

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Email: peta.clode-at-uwa.edu.au
Name: Peta Clode

Organization: University of Western Australia

Title-Subject: [Filtered] Diamond knife - 35 or 45deg?

Message: We have mistakenly ordered a Diatome 35deg ultra diamond knife instead of a 45deg ultra
diamond knife and I am wondering if the 35deg knife will be suitable for what I need? I particularly
wish to cut high quality 80-250nm thick sections of epoxy-embedded biological cells/tissues for TEM.

Thanks!

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Dear Peta,

if you visit " https://www.diatomeknives.com/ "
(not knowing which brand you have purchased, Disclaimer: No affiliation, no financial Interest, only 34 years satisfied customer of these knives)

you'll see that a diamond knife with 35° should be suitable for your task ("Ultra 35° the diamond knife for optimized sectioning results in almost all applications.") I personally have chosen in the 1980ies to use 45° diamond knives and always was happy with them.
Unfortunately I don't have experience with 35° knives but I guess that sectioning will be as easy as with 45 ° (depending perhaps a little bit on the quality of resin [polymerization grade soft-medium-hard] you use).

A [Diatome].pdf brochure you can find at: https://www.diatomeknives.com/knives/pdf/ultra_flyer_A4_USA_0306.pdf
There you can find:

CITATION:
{ {ultra 35°:
In 1989 J. C. Jésior (Ref. Jésior) demonstrated considerably reduced compression, smoother section surfaces and improved structural preservation thanks to the use of our ultra 35° knives.
In the meantime, a large number of scientists have recognized the advantages of 35° knives, in particular for sectioning Lowicryls and non-homogenous specimens, as well as non-decalcified bone, dental materials, etc.
The ultra 35° knives are perfect for sectioning relatively soft materials research specimens including metals and polymers, as well as hard specimens such as semi-conductors, super conducting oxides, catalysts, nano-crystalline ceramics, etc (Refs. Mahon, Glanvill, Swab, Quintana, Maniette, Schubert-Bischoff) END of CITATION } }

If you nevertheless are a bit hesitant about your knife's properties contact either your vendor for more information (brochures) or even contact Mr. GNAEGI Helmut at DIATOME AG, CH [Diamond knives for electron microscopy], e-mail: helmut.gnaegi-at-diatome.ch. HE would be the best source of information I can imagine....

Best wishes and good luck,

Wolfgang

Wolfgang MUSS
SALK-LKH (Gen. Hospital) and PMU (Private Paracelsus Medical University) SALZBURG
SALZBURG
AUSTRIA/EUROPE
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An: Muß Wolfgang
Betreff: [Microscopy] Diamond knife - 35 or 45deg?

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We have mistakenly ordered a Diatome 35deg ultra diamond knife instead of a 45deg ultra
diamond knife and I am wondering if the 35deg knife will be suitable for what I need?
I particularly wish to cut high quality 80-250nm thick sections of epoxy-embedded biological cells/tissues for TEM.

Thanks!

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I don't have a Tecnai TEM but have a similar problem with our JEOL microprobe. The chamber needs about 1.2 vent cycles to come up to atmosphere. I push the vent button and wait, then press the evac button until the pumps start and then press vent again.

We do have an FEI Quanta SEM. I think the software runs it through a vent cycle anytime we press vent.

We found that there was about 20 seconds of extra venting whenever we vented our chamber. That chewed through extra tanks of N2. The service engineer was able to reset the vent timer in the vacuum controls reduce the waste. I suppose a similar timer exits for the Tecnai in case you want to lengthen the time.

Warren

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Title-Subject: [Filtered] Venting TECNAI Gun

Message: Hello everybody..

I operate a TECNAI T12 SPIRIT transmission electron microscope. In order to replace the filament, you have to vent the electron gun (thermoionic), according to the FEI operating manual, before lifting up the gun cover.

The problem is, when I vent the gun it doesn't vent completely to the atmospheric pressure (log 99), it keeps holding some vacuum (log 95) that prevents me from lifting up the gun cover to change the filament.

Has anybody ever experienced anything like that?

Regards,

Ahmad Ashkhaibi

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It is possible that pirani gauges are out of calibration.

There will be a pot somewhere to tweak the gauge calibration - probably best to get a service engineer to check and do this.

Larry Stoter

} On 25 Nov 2014, at 14:48, wesaia-at-iastate.edu wrote:
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} I don't have a Tecnai TEM but have a similar problem with our JEOL microprobe. The chamber needs about 1.2 vent cycles to come up to atmosphere. I push the vent button and wait, then press the evac button until the pumps start and then press vent again.
}
} We do have an FEI Quanta SEM. I think the software runs it through a vent cycle anytime we press vent.
}
} We found that there was about 20 seconds of extra venting whenever we vented our chamber. That chewed through extra tanks of N2. The service engineer was able to reset the vent timer in the vacuum controls reduce the waste. I suppose a similar timer exits for the Tecnai in case you want to lengthen the time.
}
} Warren
}
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} Subject: [Microscopy] viaWWW:Venting TECNAI Gun
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} Title-Subject: [Filtered] Venting TECNAI Gun
}
} Message: Hello everybody..
}
} I operate a TECNAI T12 SPIRIT transmission electron microscope. In order to replace the filament, you have to vent the electron gun (thermoionic), according to the FEI operating manual, before lifting up the gun cover.
}
} The problem is, when I vent the gun it doesn't vent completely to the atmospheric pressure (log 99), it keeps holding some vacuum (log 95) that prevents me from lifting up the gun cover to change the filament.
}
} Has anybody ever experienced anything like that?
}
} Regards,
}
} Ahmad Ashkhaibi
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Message: We are looking for a postdoctoral researcher, possibly at a senior level, to be employed by
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Email: zoom-at-snu.ac.kr
Name: Ohkyung Kwon

Organization: SNU

Title-Subject: [Filtered] serial sections from softwood

Message: Hello,

I am planning to build 3D images from serial sections of pine.

1. How difficult to obtain serial sections (100 nm thick) of softwood (Pine)?

2. I heard that it is very hard to make thin sections without wrinkles.
Is that the main problem to prepare thin sections of softwood?

Please advice me for sample preparation tips.
Thank you very much for your advice in advance.

Best regards,
Ohkyung

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It largely depends on which orientation you want to look at and what thickness of stack you want to build up. Commonly we produce serial sections for light microscopy at around 25 µm thick from the radial or longitudinal faces, without any embedding using a sledge microtome and disposable steel blades. If you are using green sapwood then attempt to keep it wet as a swollen cell wall cuts better than a dry one. Heartwood may need some rewetting. A rougher method is to boil the blocks in water until they sink. Autoclaving in water works too. You can of course cut thinner, particularly if you embed and I'll discuss that further if you ask. Transverse (cross) sections may break up more but they can be prepared..
We commonly rewet the wood by vacuum impregnation with water (set up a vacuum desiccator so that you can pull a rough vacuum [water pump] but allowing an entry at the neck to allow water to be admitted. Load blocks into a glass of plastic beaker, allowing space for swelling and weigh down to prevent them from floating when water is admitted. Pull a vacuum, say for 10 minutes, suck water in, let vacuum go, leave to soak for a while). If the blocks are properly saturated they'll sink.
Cutting: If you use a high knife angle the sections will curl and you'll induce kinks in the walls (compression fractures, slip planes if you like, visible under polarised light) and if you have a too low angle you may end up cutting thick and thin, alternate sections. I use between 6° and 11°.
Even at a low angle some curling off the knife occurs and a small wetted paintbrush (like used for watercolours) can be used to assist the sections to remain flatter. There is also the possibility of cutting the sections as a book, a technique I came up with in the 1970s in an idle moment, when cutting a bulk of serial sections. In this method, if I recollect I was cutting radial longitudinal sections about 25 µm thick from rewetted pine sapwood blocks (5 x 50 x 25 mm) but ran the knife almost to the block end, but left a corner uncut, so that the sections stayed in order. When this was done the earlier sections in the book stopped the later sections from curling when they were cut.
Any further advice please ask or skype.

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Email: zoom-at-snu.ac.kr
Name: Ohkyung Kwon

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Title-Subject: [Filtered] serial sections from softwood

Message: Hello,

I am planning to build 3D images from serial sections of pine.

1. How difficult to obtain serial sections (100 nm thick) of softwood (Pine)?

2. I heard that it is very hard to make thin sections without wrinkles.
Is that the main problem to prepare thin sections of softwood?

Please advice me for sample preparation tips.
Thank you very much for your advice in advance.

Best regards,
Ohkyung

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On Nov 17, 2014, at 6:53 PM, zaluzec-at-microscopy.com wrote:

} Email: ravithakkar-at-vet.k-state.edu
} Name: Ravi
}
} Title-Subject: [Filtered] LN2 Filling system
}
} Message: Hi,
} I am looking for automated LN2 filling/ transfer system for EDS
} installed on scopes. Kindly share some suggestions based on your
} experience.
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Dear Ravi,
When we tried that at the HVEM, we had a problem when the system
failed open circuit. This caused LN2 to flow continually, and if
someone had not entered the scope room for an unrelated reason late
Friday, LN2 would have continued to flow all weekend. The excess LN2
caused the dewar bottom to become convex, and our shop had to make a
tool that we could use to return the dewar to its proper shape.
Fortunately, the instrument recovered and was usable for many years
after the event. After that experience, we filled the dewar manually
every Friday--the LN2 lasted more than a week in our set-up.
Yours,
Bill

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Title-Subject: [Filtered] Job Opportunity - Electron Microscopy and Imaging Manager - France

Message: An international pharmaceutical group, leader in its field of research, is seeking for its
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As part of the R&D Department, you will manage the Transmission Electron Microscope and Imaging Unit
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Hi Vickie!

Sorry for replying so late. I never had problems with polymerization at the tip of capsules.
Your problem looks more like uncomplete dehydration and since your material is pelleted, it affects only the tip.
One way to verify this hypothesis is to cure the same capsule with the same resin but without material.
If it polymerizes, then the problem is your material (probably incomplete dehydration).

Regards
Stephane

--------------------------------------------
On Tue, 11/18/14, zaluzec-at-microscopy.com {zaluzec-at-microscopy.com} wrote:

Subject: [Microscopy] viaWWW:Uneven Embed 812 Polymerization
To: nizets2-at-yahoo.com
Date: Tuesday, November 18, 2014, 3:41 AM

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Email: vakimler-at-med.wayne.edu
Name: Vickie Kimler

Organization: Wayne State University

Title-Subject: [Filtered] Uneven Embed 812 Polymerization

Message: Hello All,
How can I correct the uneven polymerization of my resin in
conical tip
"bottleneck" Beem capsules?

The tips where my pelleted cells are situated, are still
soft, while the
top of the block (base) is very hard.

I know my mistake that I did not use a metal heat sink on
the aspect of
the tips.

Can I continue to polymerize them in the 70 degree C oven
with a heat
sink without destroying interrelationships between
organelles? They have
been out of the oven a few days now, of course.

Any other advice?
Thanks,
Vickie

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Email: david-at-nanocraft.us
Name: David Shimmin

Organization: NanoCraft Electron Microscopy Services

Title-Subject: [Filtered] Leitz RMS Objective Labeling

Message: I am shopping for objective lenses for a Leitz Ergolux, with 20mm thread and 160mm tube
length. I am finding very little information on the internet to describe the various lens class
designations. For example "NPL", "L", "HL", "EF". In particular, I would like to know the
characteristics of the HL type. Is there a comprehensive document out there? We are talking about
lenses made in the 80's and 90's. Thank you!

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11, 17 -- From microscopylistserver-noreply-at-microscopy.com Wed Dec 3 08:22:57 2014
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Thanks to everybody contributed to the discussion!
davide

~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~~~~~~~
Davide Cristofori

direct phone = 041.234.67.26
e-mail = dcristofori[at]unive[dot]it

Università Ca' Foscari Venezia
Dipartimento di Sc. Molecolari e Nanosistemi
Via Torino, 155b I-30172 Mestre (VE) Italy
~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~~~~~~~

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Hello All,

How long will DNA remain stable on a grid before rotary shadowing?

Can I spread and collect the DNA by Kleinschmidt and the modified
Kleinschmidt cytochrome C droplet method a week before coating without
loss of detail?

The purpose of the project is simply to identify if the strands are
circular, branched, linear. Not so important to have high resolution.

Karen Kelley

==============================Original Headers==============================
7, 37 -- From vau-at-ufl.edu Tue Dec 9 05:24:55 2014
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7, 37 -- From: "Kelley,Karen L" {vau-at-ufl.edu}
7, 37 -- To: "Microscopy-at-Microscopy.com" {Microscopy-at-Microscopy.com}
7, 37 -- Subject: how long DNA good on grid without coating?
7, 37 -- Thread-Topic: how long DNA good on grid without coating?
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Dear Karen,
I regularly put DNA on carbon membranes with no over-coat protection - I
have found them to be stable indefinitely, as far as their contour on the
film is concerned. They are bound quite tightly - one usually cannot even
wash them off!
Regards,
Larry Scipioni

-----Original Message-----
X-from: vau-at-ufl.edu [mailto:vau-at-ufl.edu]
Sent: Tuesday, December 09, 2014 6:43 AM
To: LES-at-ZSGENETICS.COM

Hello All,

How long will DNA remain stable on a grid before rotary shadowing?

Can I spread and collect the DNA by Kleinschmidt and the modified
Kleinschmidt cytochrome C droplet method a week before coating without loss
of detail?

The purpose of the project is simply to identify if the strands are
circular, branched, linear. Not so important to have high resolution.

Karen Kelley

==============================Original Headers==============================
7, 37 -- From vau-at-ufl.edu Tue Dec 9 05:24:55 2014 7, 37 -- Received: from
smtp.ufl.edu (smtp-prod03.osg.ufl.edu [128.227.74.219])
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9 Dec 2014 06:24:51 -0500 7, 37 -- From: "Kelley,Karen L" {vau-at-ufl.edu} 7,
37 -- To: "Microscopy-at-Microscopy.com" {Microscopy-at-Microscopy.com} 7, 37 --

Dear Rebecca,

The wording for the advert is fine. Apart from the local paper, will
this be advertised elsewhere?

Two suggestions:

1. The Microscopy Society of America Listserver
(Microscopy-at-microscopy.com) is a free email contact list. Many jobs are
advertised there (especially for universities), so may be worth putting
something there.

2. The Institute of Physics (www.iop.org) has a wide readership.

Thanks for taking care of this.

Best, Jon

==============================Original Headers==============================
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OK, so this is going to sound crazy to some of you, but let me have a minute of your valuable time.

Recently the California Community College Chancellor's Office authorized a pilot program to let California community colleges offer bachelor's degrees.

Currently about 35/79 districts have expressed an interest in this program. Here at Delta, we are interested in doing it.

We have to prepare an application to be submitted by Dec. 19, 2014 explaining how we would structure our program, classes to offer, etc.

We already have a lot of EM courses. To raise our program to the BA level we will probably need more. That's where you can help. What kind of curriculum should be part of a BA degree in microscopy?

Keep in mind we have two subprograms, one biological, one materials. Some classes might be the same for both, some might be only good for one side.

In addition to classic microscopy classes (maybe you should tell me what these should be too) what other training would a student need to present him/her self to your lab with a new BA in hand and be competitive for joining your lab?

This all could end up as pie in the sky if we are not selected as one of the 15 to be part of the pilot program, but its a good thing for me to think about in any case.

Thanks

Jon

Jonathan Krupp
Applied , Business & Technology
San Joaquin Delta College Science
5151 Pacific Ave.
Stockton, CA 95207
209-954-5284
jkrupp-at-deltacollege.edu

Find us on Facebook -at-
Electron Microscopy at SJ Delta College

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Email: sbarlow-at-mail.sdsu.edu
Name: Steve Barlow

Organization: SDSU EM Facility

Title-Subject: [Filtered] SEM: student quick fix of cultured cells

Message: Listers

I would like to have undergrad students quickly fix 3T3 culture cells
growing on coverslips for SEM examination. I want the students to carry out
these steps as part of an introduction to using the light microscopes, SEM,
and TEM in a lab session where they photograph cultured cells labelled with
fluorescent markers, cells on the SEM, and a slice of cultured cells or T4
negatively stained bacteriophage in the TEM.

Unfortunately, there is very little time available for me to squeeze it
into the lab session that would be available to the students. Furthermore,
there are 140 students in the 2hr 45 minute lab sessions (5-6 session over
3 days depending on enrollment), and only a small hood. I would like to
avoid using osmium tetroxide because of the safety issues, and can't
critical point dry that many samples (working in pairs is still almost 70
samples).

A preliminary attempt, designed for speed and simplicity:
-Rinse cells in room temp PBS
- Fix in 4% formaldehyde, 2% glutaraldehdye in PBS for 10 minutes at room
temp.
-Rinse in room temp PBS
-dehydrate in ethanol: 50%, 75%, 95%, 100% 3x, each -at-5 min
-air dry from 100% ethanol, or from HMDS (after 100% EtOH, 50:50 EtOH HMDS,
100% HMDS 2x each -at- 5 min, then air dry).

Cell are not happy in this preparation, looking either like outlines of
where the cells were, or flattened and full of holes in their plasma
membranes. Monitoring the cells suggests the problems are occurring after
the start of the ethanol series.

Any suggestions on ways for the students to better preserve these cells
within the time and equipment constraints would be appreciated.

Steve

--
Steven Barlow, Ph.D.
Director, EM Facility/ SDSU Biology
5500 Campanile Drive MC-4614
San Diego, CA 92182-4614
phone: (619) 594-4523
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Steve,

When I've done cultured cells:
Grow on coveslips - first sputter coat the coverslips on both sides for
conductivity. 1 min max - this won't interfer with light microscopy.
Fix with 1 - 1.25% glut in preferred buffer (0.1 M PO4 worked fine) plus
1% monomeric tannic acid. The TA helps with the membrane hole. But! Fix
for 1 hour at room temp. 10 minutes is too short.

deH2O through EtOH starting at 30%, maybe lower depending on the cells
30-50-70-80-90-95-3x100
then to HMDS, but! use more intermediate steps.
2:1, 1:1, 1:2 EtOH:HMDS, or even 3:1, 3:2, 1:1, 2:3, 1:3 EtOH:HMDS
Then 3x100% HMDS
5 min steps is generally enough if the cells are a monolayer, but I'd
use 10 min for the transition to HMDS and the HMDS steps.
Air dry either room temp or at 60 deg C - which works better depends on
the cells, just have to try. (Sometimes 37 deg or 45 deg is best.)
This takes 1 hr or more.

It's just possible that t-Butyl alcohol would work. After the EtOH
series, go 2:1, 1:1, 1:2 EtOH:tBuOH, 3x100% tBuOH, let solidify (freezes
below 25 deg C) and vacuum sublimate. Tobias Baskin, May 2014 Microscopy
Today. Leave just enough tBuOH to cover the cells before letting freeze.
Also takes ~hr, maybe more.
I have not tried this with cells, though. But, tBuOH is much less toxic
than is HMDS.

Not doable in one lab session.

Phil

} Email: sbarlow-at-mail.sdsu.edu
} Name: Steve Barlow
}
} Organization: SDSU EM Facility
}
} Title-Subject: [Filtered] SEM: student quick fix of cultured cells
}
} Message: Listers
}
} I would like to have undergrad students quickly fix 3T3 culture cells
} growing on coverslips for SEM examination. I want the students to carry out
} these steps as part of an introduction to using the light microscopes, SEM,
} and TEM in a lab session where they photograph cultured cells labelled with
} fluorescent markers, cells on the SEM, and a slice of cultured cells or T4
} negatively stained bacteriophage in the TEM.
}
} Unfortunately, there is very little time available for me to squeeze it
} into the lab session that would be available to the students. Furthermore,
} there are 140 students in the 2hr 45 minute lab sessions (5-6 session over
} 3 days depending on enrollment), and only a small hood. I would like to
} avoid using osmium tetroxide because of the safety issues, and can't
} critical point dry that many samples (working in pairs is still almost 70
} samples).
}
} A preliminary attempt, designed for speed and simplicity:
} -Rinse cells in room temp PBS
} - Fix in 4% formaldehyde, 2% glutaraldehdye in PBS for 10 minutes at room
} temp.
} -Rinse in room temp PBS
} -dehydrate in ethanol: 50%, 75%, 95%, 100% 3x, each -at-5 min
} -air dry from 100% ethanol, or from HMDS (after 100% EtOH, 50:50 EtOH HMDS,
} 100% HMDS 2x each -at- 5 min, then air dry).
}
} Cell are not happy in this preparation, looking either like outlines of
} where the cells were, or flattened and full of holes in their plasma
} membranes. Monitoring the cells suggests the problems are occurring after
} the start of the ethanol series.
}
} Any suggestions on ways for the students to better preserve these cells
} within the time and equipment constraints would be appreciated.
}
} Steve
}
} --
} Steven Barlow, Ph.D.
} Director, EM Facility/ SDSU Biology
} 5500 Campanile Drive MC-4614
} San Diego, CA 92182-4614
} phone: (619) 594-4523
} fax: (619) 594-5676
} email: sbarlow-at-mail.sdsu.edu
} web: www.sci.sdsu.edu/emfacility
}
} Login Host: 146.244.234.237
} Listserver Email Form V - 20120416
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}

--
Philip Oshel
Microscopy Facility Supervisor
Biology Department
024C Brooks Hall
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576

==============================Original Headers==============================
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Jon;
My thought, as one who has hired numerous staff for industry, is that a BS degree would be desirable, but most hiring requisitions for an engineer will not be for doing technician work, even if that might be a good idea. For materials microscopy, you would probably need to extend it to be a Materials Science degree. However, I do recall one of the best people our lab at Intel ever hired was a young lady with a BS degree in metallography from the Max Planck institute. She is still in the Bay area and works for JEOL, so you could consult with her and find out what their curriculum was.

John Mardinly, ASU

-----Original Message-----
X-from: jkrupp-at-deltacollege.edu [mailto:jkrupp-at-deltacollege.edu]
Sent: Tuesday, December 09, 2014 5:42 PM
To: John Mardinly

OK, so this is going to sound crazy to some of you, but let me have a minute of your valuable time.

Recently the California Community College Chancellor's Office authorized a pilot program to let California community colleges offer bachelor's degrees.

Currently about 35/79 districts have expressed an interest in this program. Here at Delta, we are interested in doing it.

We have to prepare an application to be submitted by Dec. 19, 2014 explaining how we would structure our program, classes to offer, etc.

We already have a lot of EM courses. To raise our program to the BA level we will probably need more. That's where you can help. What kind of curriculum should be part of a BA degree in microscopy?

Keep in mind we have two subprograms, one biological, one materials. Some classes might be the same for both, some might be only good for one side.

In addition to classic microscopy classes (maybe you should tell me what these should be too) what other training would a student need to present him/her self to your lab with a new BA in hand and be competitive for joining your lab?

This all could end up as pie in the sky if we are not selected as one of the 15 to be part of the pilot program, but its a good thing for me to think about in any case.

Thanks

Jon

Jonathan Krupp
Applied , Business & Technology
San Joaquin Delta College Science
5151 Pacific Ave.
Stockton, CA 95207
209-954-5284
jkrupp-at-deltacollege.edu

Find us on Facebook -at-
Electron Microscopy at SJ Delta College

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Microsc. Microanal.["Microscopy and Microanalysis"] 20, 1348-1355, 2014, "Extracellular Matrix Reorganization during Cryo Preparation for Scanning Electron Microscope Imaging of Staphylococcus aureus Biofilms" (doi: 10:1017/S143192761401277X)

Comparison of Drying: In air; plunging in Liquid Nitrogen(LN2), plunging in liquid ethane cooled in LN2, and high pressure freezing in LN2 - followed by low temp sublimation of the frozen HOH. I suggest this to emphasize the value of high tech freezing methods followed by sublimation under vacuum, AND cryo transfer equipment to move the specimen onto the SEM stage - itself held at regulated low temperatures. I lack these as well, so the article can provide a student with the difference between what s/he gets comparted with what is now appropriate in the primary literature.

I, and others, have used in sequence, OsO4 gas (from 1ml 10% OsO4 in HOH + 4ml HOH) to stabilize membranes in a closed '"Ball" jar' in the hood for 1-2hr; followed by an appropriate buffered fixative for 10-60min+ at 4oC. E.g., when I required a urinary bladder to be fixed at volume, I resorted to the following. Immerse bladder at volume in buffered paraldehyde for 10 minutes at 4oC. This poisons the musculo-neural system so that when the urine is released, the same volume of fixative can replace it to the same volume also at 4oC. I would then leave the bladder to fix overnight before continuing processing. The same works for TEM preps, but one must be aware that a rabbit urinary bladder wall becomes quite thin when distended.

A colleague, and, as reported, colleagues of his, often used OsO4 gas to stabilize his bacterial preps, either for thin sectioning or whole cell TEM studies such as negative staining.

Hope this helps,

Fred Monson

Frederick C. Monson, PhD
Technical Director
Center for Microanalysis and Imaging, Research and Training (CMIRT)
West Chester University of Pennsylvania
Schmucker Science South - Room SSS-024
MailDrop: Geology-Astronomy
750 South Church Street
West Chester, PA, 19383
610-738-0437
fmonson-at-wcupa.edu

-----Original Message-----
X-from: oshel1pe-at-cmich.edu [mailto:oshel1pe-at-cmich.edu]
Sent: Wednesday, December 10, 2014 11:00 AM
To: Monson, Frederick

Steve,

When I've done cultured cells:
Grow on coveslips - first sputter coat the coverslips on both sides for conductivity. 1 min max - this won't interfer with light microscopy.
Fix with 1 - 1.25% glut in preferred buffer (0.1 M PO4 worked fine) plus 1% monomeric tannic acid. The TA helps with the membrane hole. But! Fix for 1 hour at room temp. 10 minutes is too short.

deH2O through EtOH starting at 30%, maybe lower depending on the cells
30-50-70-80-90-95-3x100
then to HMDS, but! use more intermediate steps.
2:1, 1:1, 1:2 EtOH:HMDS, or even 3:1, 3:2, 1:1, 2:3, 1:3 EtOH:HMDS Then 3x100% HMDS
5 min steps is generally enough if the cells are a monolayer, but I'd use 10 min for the transition to HMDS and the HMDS steps.
Air dry either room temp or at 60 deg C - which works better depends on the cells, just have to try. (Sometimes 37 deg or 45 deg is best.) This takes 1 hr or more.

It's just possible that t-Butyl alcohol would work. After the EtOH series, go 2:1, 1:1, 1:2 EtOH:tBuOH, 3x100% tBuOH, let solidify (freezes below 25 deg C) and vacuum sublimate. Tobias Baskin, May 2014 Microscopy Today. Leave just enough tBuOH to cover the cells before letting freeze.
Also takes ~hr, maybe more.
I have not tried this with cells, though. But, tBuOH is much less toxic than is HMDS.

Not doable in one lab session.

Phil

} Email: sbarlow-at-mail.sdsu.edu
} Name: Steve Barlow
}
} Organization: SDSU EM Facility
}
} Title-Subject: [Filtered] SEM: student quick fix of cultured cells
}
} Message: Listers
}
} I would like to have undergrad students quickly fix 3T3 culture cells
} growing on coverslips for SEM examination. I want the students to
} carry out these steps as part of an introduction to using the light
} microscopes, SEM, and TEM in a lab session where they photograph
} cultured cells labelled with fluorescent markers, cells on the SEM,
} and a slice of cultured cells or T4 negatively stained bacteriophage in the TEM.
}
} Unfortunately, there is very little time available for me to squeeze
} it into the lab session that would be available to the students.
} Furthermore, there are 140 students in the 2hr 45 minute lab sessions
} (5-6 session over
} 3 days depending on enrollment), and only a small hood. I would like
} to avoid using osmium tetroxide because of the safety issues, and
} can't critical point dry that many samples (working in pairs is still
} almost 70 samples).
}
} A preliminary attempt, designed for speed and simplicity:
} -Rinse cells in room temp PBS
} - Fix in 4% formaldehyde, 2% glutaraldehdye in PBS for 10 minutes at
} room temp.
} -Rinse in room temp PBS
} -dehydrate in ethanol: 50%, 75%, 95%, 100% 3x, each -at-5 min -air dry
} from 100% ethanol, or from HMDS (after 100% EtOH, 50:50 EtOH HMDS,
} 100% HMDS 2x each -at- 5 min, then air dry).
}
} Cell are not happy in this preparation, looking either like outlines
} of where the cells were, or flattened and full of holes in their
} plasma membranes. Monitoring the cells suggests the problems are
} occurring after the start of the ethanol series.
}
} Any suggestions on ways for the students to better preserve these
} cells within the time and equipment constraints would be appreciated.
}
} Steve
}
} --
} Steven Barlow, Ph.D.
} Director, EM Facility/ SDSU Biology
} 5500 Campanile Drive MC-4614
} San Diego, CA 92182-4614
} phone: (619) 594-4523
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}

--
Philip Oshel
Microscopy Facility Supervisor
Biology Department
024C Brooks Hall
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576

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In less than 24 hours I have received at least 18 replies to my question about what should be included in an undergraduate microscopy curriculum.

If I don't get a chance to reply to everyone individually, this is a big shout out of thanks for the ideas.

You have given me a wealth of ideas to check. Keep them coming.

Jon

Jonathan Krupp
Applied Science, Business & Technology
San Joaquin Delta College
5151 Pacific Ave.
Stockton, CA 95207
209-954-5284
jkrupp-at-deltacollege.edu

Find us on Facebook -at-
Electron Microscopy at SJ Delta College

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Hi, Jon

What a wonderful opportunity!

If you are doing a lot of EM, then I am assuming that you are also including typical elemental analyses such as EDS, WDX, EBSD, etc. With the rise in both hybridized and interfaced systems for IR (both FT-IR and NIR) and Raman I would also encourage you to add a course on molecular analyses.

In terms of expanding the general microscopy component, definitely courses in AFM sample prep and analysis (important now in both bio and materials). On the materials side, it would be helpful to have surface analysis such as phase shifting and white light interferometry, with perhaps either modules and/or additional courses on tribology and nanoindentation.

Hope this is helpful.

Good hunting!

Barbara Foster, President & Chief Consultant
Microscopy/Microscopy Education*
www.MicroscopyEducation.com

P. S. Have you taken part yet in MME's "20 Really Quick Quesions"? The deadline has been extended to this Saturday, Dec. 13! See www.MicroscopyEducation.com for details.

At 10:32 AM 12/11/2014, jkrupp-at-deltacollege.edu wrote:

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I find it interesting that the discussion is about a course of study to lead to a BA but all the topics I'm seeing seem to be electron microscopy. While these are all excellent ideas, let's not forget about LM.

No course of study would be complete without training in:
Photomicroscopy and macrophotography,
Phase contrast for both the biological and material science side,
Polarized light microscopy including immersion methods and optical crystallography,
Reflected light microscopy including metallography and hardness testing.

Of course, I'm partial to fiber and hair microscopy, microchemical testing and fusion methods, but these may not have sufficient impact in a general study.

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11, 30 -- Subject: Re: Undergraduate curicculum?
11, 30 -- Thread-Topic: Undergraduate curicculum?
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I thought much of the difference between an associate's degree and a BA or BS degree was about a broader education not just more technical topics. I have been involved twice this year with hiring someone to operate SEMs and other equipment. The position involves operation and training, but it also involves lots of materials science which I do not suppose would come with just an associate's degree. We also look for excellent communication skills which may or may not be part of a community college degree.

On the technical side, we have a strong interest in EDS and EBSD to go along with the materials science.

FWIW, 17 of our 19 candidates had PhD degrees although we only called for a BS or MS degree.

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Email: jhyun-at-gatan.com
Name: John Hyun

Organization: Gatan

Title-Subject: [Filtered] EELS & EFTEM Analysis Training School 2015

Message: If you are interested in advancing your EELS and EFTEM skills, we invite you to register
for the April 2015 EELS & EFTEM Analysis Training School, April 14-17, 2015, Gatan R&D
Headquarters, Pleasanton, CA USA.

This course reviews the basic theory and practice of EELS imaging and analysis in the TEM, but its
main emphasis is on practical techniques, optimum deployment of Gatan hardware and software systems,
and advanced EELS and EFTEM applications. Some prior experience with EELS, EFTEM, and Gatan systems
is recommended, as is a good familiarity with TEM/STEM instrumentation and techniques.

You can register online at:
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Email: ravi.thakkar369-at-gmail.com
Name: Ravi

Title-Subject: [Filtered] Gamma settings in Gatan Digital Micrograph

Message: Hi,
I am interested to analyse gold nanoparticle on RBC surface.
I have used Olympus AnalySIS before, it has gamma setting to visualize nanoparticle on RBC surface.
Does Gatan digital micrograph has such option.?

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10, 17 -- From microscopylistserver-noreply-at-microscopy.com Thu Dec 11 16:51:31 2014
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Dear Ravi,
You'll find gamma, brightness and contrast sliders by selecting the menu
item Window/Floating Windows/Display Control. ImageJ is a good place to do
this as well (in my opinion a better place, actually).
Regards,
Larry Scipioni

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Title-Subject: [Filtered] Gamma settings in Gatan Digital Micrograph

Message: Hi,
I am interested to analyse gold nanoparticle on RBC surface.
I have used Olympus AnalySIS before, it has gamma setting to visualize
nanoparticle on RBC surface.
Does Gatan digital micrograph has such option.?

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18, 44 -- From les-at-zsgenetics.com Fri Dec 12 08:56:58 2014
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Email: lamiller-at-illinois.edu
Name: Lou Ann Miller

Organization: Materials Research Laboratory, University of Illinois

Title-Subject: [Filtered] Job opening for an experienced Electron Microscopist

Message: Research Scientist(s) position
Please see the link below for the job information and to apply electronically online:

https://jobs.illinois.edu/academic-job-board/job-details?jobID=36125&job=research-scientist-materials-research-laboratory-a1300472

*Note for those unfamiliar with University jobs here, a common question: Is the job for only one
year with the "12 month appointment"?

All academic positions at the University are appointed on a 9- or 12-month service basis, eligible
for annual renewal based upon mutual agreement and the annual performance review process. This
Academic Professional position is a regularly recurring staff scientist role not subject to grant
funding. See http://ap.illinois.edu for more information.

{ { { { { { { { { { { { { { { { { {} } } } } } } } } } } }
Lou Ann Miller
Biological Electron Microscopy
Frederick Seitz Materials Research Laboratory
Lab Room 125 Office 374 MRL
104 South Goodwin Ave
Urbana, IL 61801
Campus mail code: MC-230

217-244-1567
lamiller-at-illinois.edu
http://mrl.illinois.edu

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All,

The deadlines for abstract submission and early-bird registration for AMAS XIII, the 13th biennial Australian Microbeam Analysis Symposium, have been extended to 23 December 2014.

So it's not too late to submit your paper and join us this (Australian) Summer in beautiful Hobart, Tasmania, on 11-13 February 2015, with Pre-Meeting Workshops 9-10 Feb, 2015. AMAS is also still accepting applications for student travel bursaries for the conference.

The AMAS symposia provide a forum to discuss and share ideas on advances, trends and challenges in microanalysis and imaging with national and international leaders in the field, with an emphasis on practical solutions and applications. A range of exciting speakers has already been confirmed, including European and American attendees visiting Australia for the first time.

Please see our website

http://www.microscopy.org.au/amas/amas13/

for more information such as second circular, invited speakers, workshop program, registration,
accommodation, and student travel bursary information.

Please feel free to forward this email to your colleagues and anyone you think might be interested in attending.

Apologies for cross-posting.

Many thanks,

Karsten

AMAS XIII Co-Chair

Dr Karsten Goemann
Research Fellow, Electron Microscopy & X-Ray Microanalysis
Central Science Laboratory, University of Tasmania
Mail: Private Bag 74, Hobart TAS 7001, Australia
Location: Rooms 254-256, Chemistry Building, Dobson Road, Sandy Bay TAS 7005, Australia
T: +61 (0)3 6226 2146 | F: +61 (0)3 6226 2494 | M: +61 (0)407 101 990
www.utas.edu.au/research/central-science-laboratory
CRICOS 00586B

University of Tasmania Electronic Communications Policy (December, 2014).
This email is confidential, and is for the intended recipient only. Access, disclosure, copying, distribution, or reliance on any of it by anyone outside the intended recipient organisation is prohibited and may be a criminal offence. Please delete if obtained in error and email confirmation to the sender. The views expressed in this email are not necessarily the views of the University of Tasmania, unless clearly intended otherwise.

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Dear fellow Microscopists,

It must a be a glitch reposting something I posted a long time ago. maybe a year ago.
A job opening posted yesterday, I did not put out a job posting yesterday. Please disregard this.

However we do have an opening in our â€œcleanâ€ microfab laboratory, you may want to look on our job board for that.

Sorry, I posted nothing, so do not know how this happened.

Lou Ann

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10, 41 -- From lamiller-at-illinois.edu Sun Dec 14 07:28:07 2014
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Colleagues

The Job posting from UIUC which appeared on the listserver
yesterday is abit over a year old. It originally was posted
on 10/23/13.

I am guessing that a copy of the original message was stuck in
an internet blackhole somewhere and on a system reboot somewhere
on the UIUC campus the message got resent.

Apologies to all, but the MListerver computer
registered it as a new posting.

Nestor
Your Friendly Neighborhood SysOp

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Email: er12-at-rice.edu
Name: Emilie Ringe

Organization: Rice University

Title-Subject: [Filtered] 2 TEM FIB Tech/postdoc Positions Available at Rice University, TX, USA

Message: The new Electron Microscopy Center (EMC) at Rice University in Houston, TX,
USA is opening in 2015, and we are looking to fill two positions to perform
both student instrument training and to participate in state of the art
nanomaterials research

The EMC consists of a fully equipped FEI Titan Themis3, with probe and
image correctors as well as monochromator and advanced detectors, and a FEI
Helios 660, both installed in a state-of-the art facility.

We are looking for 1 technician/postdoc specializing in S/TEM and EELS, and
1 technician/postdoc specializing in SEM/FIB and TEM sample preparation.
Anticipated start date is March 2015 with an initial appointment for 2
years and extension to a permanent position possible/desirable for
exceptional performance.

Each position will include ample time (50%) to pursue his/her own research
in collaboration with one or more EMC associated faculty, as well as a
competitive salary, plus full fringe benefits. Tool maintenance is provided
by a service contract and the employee is expected to act as the principal
liaison with the service provider. The candidate will be provided full
administrative support with respect to the instrument billing, management,
and related tasks.

As a research associate the candidate will join a research group in the new
Materials Science and NanoEngineering department at Rice University. More
information on MSNE can be found here: http://msne.rice.edu/

Ph.D. degree in materials science or equivalent field; Non-Ph.D. candidates
with significant experience may be considered. Extensive hands-on
experience in state of the art SEM/FIB and TEM required, as well as
appropriate EM sample prep techniques. Applications will be considered on
an ongoing basis until the positions is filled, and should include a CV,
name and contact info of three references, as well as a brief statement of
research interests.

Applications will be accepted on an ongoing basis, potential candidates
will be interviewed in late January. Questions and applications can be
directed to Cindy Wikes, Department coordinator (cindy.wikes-at-rice.edu), or
Emilie Ringe, faculty in charge (emilie.ringe-at-rice.edu).

Thanks!

Emilie Ringe, Ph.D.
Assistant Professor
Department of Materials Science and Nanoengineering
Department of Chemistry
Rice University
6100 Main MS-325, Houston, TX 77005
Email: er12-at-rice.edu
Department: http://msne.rice.edu/Content.aspx?id=109
Research Group: http://ringegroup.blogs.rice.edu/

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Email: jinsong-wu-at-northwestern.edu
Name: jinsong wu

Organization: Northwestern University

Title-Subject: [Filtered] Permanent Staff Position in Advanced S/TEM at Northwestern University,
Evanston, IL, USA

Message: Open Position - Permanent Scientific Staff
Northwestern University Â– NUANCE Center

NUANCE, Northwestern UniversityÂ’s Atomic and Nanoscale Characterization Experimental Center, is
seeking a Senior Research Associate (permanent scientific staff position) to oversee research,
collaboration and training related to the Scanning Transmission and Transmission Electron Microscopy
(S/TEM) facilities.

QUALIFICATIONS
The preferred candidate will have obtained a PhD in Materials Science, Physics or related physical
science/engineering field. The preferred candidate will have at least 3 years of experience in
advanced S/TEM analysis, related analytical techniques and sample preparation methods for physical
science/engineering materials. Proven publication record utilizing advanced S/TEM techniques and
applications is essential.

SKILLS
The preferred candidate will have demonstrated expertise in conventional and advanced S/TEM
techniques/skills, including: defect imaging, HREM, HAADF, electron diffraction, EDS, EELS, among
others. The candidate will have demonstrated expertise in TEM-related sample preparation skills,
including: conventional thin foil preparation and FIB/related techniques. Experience in
aberration-corrected S/TEM is highly desirable.

The candidate will have demonstrated the ability to facilitate lab sessions for hands-on course and
curricula related to conventional and advanced electron microscopy. The candidate will be capable
of using software for simulation, modeling and tomographic reconstruction, as well as other software
related advanced imaging/spectroscopy analysis. The successful candidate will be customer-oriented
and demonstrate the ability to design experiments and research programs to solve innovative problems
in atomic and nanoscale analysis and characterization.

DUTIES
The selected candidate will be responsible for training and assisting users on conventional and
advanced S/TEM, including aberration-corrected S/TEM, facilitating laboratory sessions for
microscopy courses, and conducting analysis and characterization using electron microscopes. There
are ample opportunities for independent and collaborative research utilizing advanced S/TEMs. The
selected candidate will regularly operate conventional and advanced S/TEM analysis, consult and
collaborate with users on experiment design & research in electron microscopy, sample preparation
and general lab equipment. With the expected installation of aberration-corrected S/TEM in 2015,
preference will be given to those candidates with proven experience in AC S/TEM.

Materials should be submitted as a single PDF to chad.goeser-at-northwestern.edu
Complete applications will include:

1. Introduction letter
2. Curriculum Vitae
3. Statement of plans for research (3 to 5 pages)
4. Contact information for three references (names, postal & email addresses, phone number)

Review of submissions will commence January 1, 2015 and further review will occur until the search
is closed.

Northwestern University is an Equal Opportunity, Affirmative Action Employer of all protected
classes including veterans and individuals with disabilities.

http://www.nuance.northwestern.edu/epic/senior_research_associate.html

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21, 18 -- From microscopylistserver-noreply-at-microscopy.com Mon Dec 15 17:29:03 2014
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Email: hipps-at-wsu.edu
Name: K W Hipps

Organization: Washington State University

Title-Subject: [Filtered] Used STM wanted

Message: I am looking for a used Molecular Imaging or Agilent STM. A 5500, pico plus, or Pico would
work. I could either buy the entire system, or just the STM head, chamber and interface since I own
an extra controller.

Best wishes

KW

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Email: swatkins-at-pitt.edu
Name: Simon watkins

Organization: university of pittsburgh

Title-Subject: [Filtered] JEOL negative trays

Message: Folks, a few months back we took delivery of a new JEOL 1400. As we do basic biologic TEM
with this scope we are entirely digital.. It came with 100 negative holders which are still packed
up in plastic bags and completely unused.
If you have a legitimate use for them please let me know and we will ship them to you
Happy holidays
Simon

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Email: ravi.thakkar369-at-gmail.com
Name: Ravi

Title-Subject: [Filtered] Line in diffraction image.

Message: I am getting a line in diffraction pattern for all the
samples.(https://www.flickr.com/photos/97321550-at-N08/15858529368/) I am sure this is not from the
sample. It's something to do with scope. Can Anybody here tell me how to get rid of this line.

Thanks.

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Hi Ravi,

This looks like a TEM diffraction pattern with an incorrect timing on the blanking shutter. If you are using Digital Micrograph you can look under the Camera pallet, acquisition settings and set the camera to start acquiring the image 0.1 seconds or so after the blanking shutter turns off and see if it goes away. If so, then the explanation is likely that it was due to the image acquisition starting the INSTANT the beam blanking turned off. Of course, during that instant the beam is still slewing back into place, so that faint line is actually the direct beam rastering back to it's place under the beam blocker.

Another hint is that if this is happening, the line appears stronger for shorter acquisitions and fainter for longer (though the actual intensity is the same if you count the number of counts). The time it takes for the shutter to turn off is fixed (say, 0.01 secs of something) whereas the total acquisition time varies.

A little caution, the shutter is often important to the well being of your camera, so make sure you're configuring the right setting!

Zack

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Name: Ravi

Title-Subject: [Filtered] Line in diffraction image.

Message: I am getting a line in diffraction pattern for all the
samples.(https://www.flickr.com/photos/97321550-at-N08/15858529368/) I am sure this is not from the
sample. It's something to do with scope. Can Anybody here tell me how to get rid of this line.

Thanks.

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

Dear colleagues

We are enlarging our team and are searching for a Technician. Our new colleague will devote his/her time mainly to biological sample preparation for electron microscopy.

IST Austria is a new international institute dedicated to be a world-class research center focused on the natural sciences, which is located in Klosterneuburg on the outskirts of Vienna.
Electron microscopy facility team at IST Austria provides researchers with well-maintained cutting-edge sample preparation and imaging instrumentation and modern infrastructure.

Main tasks:
ï‚§ Biological sample preparation for electron microscopy.
ï‚§ Support and training of users in the field of sample preparation techniques.
ï‚§ Care of the Electron Microscopy Facility equipment â€“ conduct operational maintenance of sample preparation equipment.

Profile of a candidate:
ï‚§ Working experience with biological sample preparation for electron microscopy observation â€“ ultrastructural immunolabeling techniques and ultramicrotomy.
ï‚§ Working experience in a research environment in the field of natural sciences (biology, chemistry).
ï‚§ Ability to work independently and team-oriented.
ï‚§ High degree of reliability, organizational and interpersonal skills.
ï‚§ Good knowledge in English.

We welcome flexible and reliable team players who are interested in working in an international environment within a diverse and dynamic working atmosphere. Possible starting date is 1.3.2015 or later.
Salary will be depending on education, qualification, and work experience.
IST Austria values diversity and is committed to equality, minimal salary 1900 â‚¬ gross/month.

To apply for this position, please send your application in one combined pdf (including CV, photo, certificates and references) by e-mail to: ludek.lovicar-at-ist.ac.at

Please feel free to forward this post to anyone who might be interested in the position. This job ad is also available at:
http://ist.ac.at/about-ist-austria/open-positions/scientific-services/

Kind regards,
Ludek Lovicar

-----------------------------------------------------
Ludek Lovicar
Electron Microscopy Facility
Scientific Services Unit
Institute of Science and Technology Austria
Am Campus 1
3400 Klosterneuburg
Austria

Phone: +43-(0)2243 9000 1066
Mobile: +43-(0)664 604 841 066
Fax: +43-(0)2243 9000-2000
Email: ludek.lovicar-at-ist.ac.at

Visit our website: www.ist.ac.at

==============================Original Headers==============================
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15, 39 -- Date: Thu, 18 Dec 2014 16:40:44 +0100
15, 39 -- Subject: Open position in the field of EM at IST Austria
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From secure-at-inc.pay1.com Thu Dec 18 16:29:54 2014
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I want to study melting and merging of two crystals under high
resolution TEM. Since we do not have an in-situ heating stage, idea is
to coat the TEM grid with sample and heat it in vacuum ( {700 degree C)
then see it under TEM. I would like to know which substrate/grid
material will be most suitable for it?
Will normal carbon or carbon/formvar coated copper grids will work or
shall i look for membranes like silicon nitride?

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

Dear Microscopy Listserver Members,

After serving the microscopy community for over 60 years, Theodore &
Christel Pella will retire from the regular workweek at the end of this
month. I'm not sure there is another couple who has served science so long.
They have greatly enjoyed serving the microscopy community. They have
watched, and in some cases been a part of that history. They attended first
EMSA, and then M&M meetings for over 50 years, even prior to forming Ted
Pella, Inc. Their story follows.

Christel started in the field as a TEM technologist, working in Sweden in
the lab of Fritiof Sjostrand at Karolinska Institute in the early 1950's.
She was part of a group that first saw cell ultrastructure.

At the same time Ted joined the fledgling Sorvall Company as a sales
engineer, and not long afterwards LKB. Ted delivered the first commercial
ultramicrotome under his arm to the Rockefeller Institute in New York,
taking a train to get there. He greatly enjoyed visiting with customers,
helping repair instruments in their labs and finding ways to help.

As Ted's responsibilities grew, LKB sent him to Sweden for further EM
training. Christel was conducting the training and they met there. Christel
emigrated to the USA and they got married in 1957. Chris continued to work,
now at Johns Hopkins but stopped to start a family until my sister and I
started school.

About that time LKB promoted Ted and the family moved to Sweden for a few
years. It proved to be a tough work transition, however, and Ted began
thinking about returning. During a vacation Ted bounced the idea off Chris:
Why don't we start our own business back in the states? Chris had a spirit
of adventure and they agreed to start it together. The family moved back to
the states, this time to Southern California where on Jan. 1, 1968 Ted
Pella, Inc. was started first in our house in Altadena, then moved a few
years later to Tustin. It was during these first years that the company
switched away from a focus on instruments to a focus on supplies, one that
has stood the test of decades.

Throughout this time Ted and Chris worked hard to listen to customer needs
and take on the products that labs used. So many products and techniques
were tried and perfected by the microscopy community over those years; so
much equipment was developed and redeveloped to suit the discoveries and
changing needs of labs.

Within a few years they rented their first office, then the one next door in
addition, and yet a third as the business slowly grew. I remember the very
long hours Ted worked when he split a 16-hour day for months; half the time
managing the business during the day and the other half working on a
composer in the early morning and evening, setting type for a catalog.

During these early years Chris opened Pelco International to better serve
the unique needs of our international customers. She grew up in Europe, was
trilingual and had sensitivity to people in other countries. She ran that
business for over 30 years and it grew into a significant sister company
until it became part of Ted Pella, Inc. in 2007. She has continued running
that internal international business until this day.

The time finally came when they wanted to own their own building; but this
was the time of the great real estate boom of the 80's and no land was
available nearby for building. The City of Redding "found" them at a trade
show and introduced them to what Northern California had to offer. Ted &
Chris moved the company to Redding in the Spring of 1987. At that time the
number of employees had grown to 19.

The company has continued to slowly grow over time as the number of labs has
grown worldwide, until now when we have over 60 employees. Many of you have
gotten to know Ted & Chris as they answered questions, partnered with you on
product development and filled hundreds of thousands of your purchase
orders. They helped sponsored many societies and visited with you, their
customers, all over the USA and all over the world.

In 2010 they passed the business on to me but continued to be actively
involved because this field is their passion, it is in their DNA. The
company is what it is today because of the foundation they established.

If any of you would like to drop me a note of congratulations to them or
recall how you know them to me at tom_pella-at-tedpella.com, we would be
honored and add it to a book we're putting together for them.

Thank you.

Best Regards,

Tom Pella
President
Ted Pella, Inc.
PO Box 492477
Redding, CA 96049
Tel: 530-243-2200
Fax: 530-243-3761
http://www.tedpella.com

==============================Original Headers==============================
21, 20 -- From tom_pella-at-tedpella.com Tue Dec 23 13:42:08 2014
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21, 20 -- From: "Tom Pella" {tom_pella-at-tedpella.com}
21, 20 -- To: {Microscopy-at-microscopy.com}
21, 20 -- Subject: Announcement: Retirement Of Theodore And Christel Pella
21, 20 -- Date: Tue, 23 Dec 2014 11:41:45 -0800
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