RE: Whole mouse snap freezing for frozen sections
Dear
Gayle,
It was
some years ago now at a different institution, but I have successfully
croysectioned whole mice on a standard Leica (mid range) cryostat - a
1090 if I remember correctly.
I used
high profile gold plated disposable blades by Sturkey, & cut at 60
um. The tissue was collected using a commercially available cryo
tape (as opposed to a histo tape such as Cryojane). I got about 2
sections per disposable blade.
The mice
were pre frozen on dry ice, prior to delivery to the cryosat, I removed
the head, limbs and tail (as only the visceral organs were of interest)
prior to mounting for sectioning.
Regards
Rob
W.
At 01:52 PM 26/06/2002 -0600, Gayle Callis wrote:
I do not cryosection whole mouse
but have learned some options.
Leica 3050 research cryostat or Leica's very large, chest freezer
cryostat
(not sure of the model, but it takes up an enormous amount of floor
space
and costs approximately $190,000 equipped with special knives). The
3050
is used with
d profile tungsten carbide tips mounted in steel backs - microtome
blades.
DDK sells these, and they are excellent.
I am not sure what thickness, but Robin Taylor indicated 25 um picked up
on
tape.
Instrumedics has a macro Cryojane unit, very pricey, but useful for
tape
transfer with bone or a specimen this large. I viewed a poster
using this
technology for sectioning whole mouse embedded in paraffin, and know it
can
be used for frozen sections. I guess in terms of pricey, any
larger
cryostat would also fit this description not to mention tungsten
carbide
knives plus their reconditioning costs, one knife would not be very
efficient.
As for how to cryosection a whole mouse, I envy the person who
successfully
does this, they must have great patience and skill. We looked whole
mouse
frozen serial sections some time ago. It was later learned that
high
resolution CCD cameras and specially labelled cells to produce
bioluminescence was far cheaper to perform T and B cell trafficking in
a
live mouse, with possibility of using the mouse again for further
testing.
Heavy sigh of relief on my part.
Obviously the right tools are very important.
At 10:20 AM 6/26/02 -0700, you wrote:
> Hi Gayle, how do you cryosection (what
thickness, what type of
>...tome?).
>Thanks in advance,
>Abizar > -----Original Message-----
Gayle Callis
MT,HT,HTL(ASCP)
Research Histopathology Supervisor
Veterinary Molecular Biology - Marsh Lab
Montana State University - Bozeman
19th and Lincoln St
Bozeman MT 59717-3610
406 994-6367 (voice mail)
406 994-4303 (FAX)
email: UVSGC@montana.edu
Robert Wadley
Senior Laboratory Manager
Flow Cytometry
Faculty of Veterinary Science
University of Sydney
Ph +61 2 9351 5827
Fax +61 2 9351 3957
http://www.usyd.edu.au/vetfac/reprogen/
RMC Gunn Building, B19, Room 212
Faculty of Veterinary Science
University of Sydney
Sydney, New South Wales, Australia, 2006
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