[Histonet] Questions on cryotome for large samples and autoradiography

[JonesLy <@t> mir.wustl.edu]

Hello -
I'm new to the list and have little experience with thin slicing of
tissues.  (Our lab does some autoradiography but usually with 1 mm or
larger slices.)  We may be getting a new scanner and to go with it plan to
purchase a cryotome.  We definitely need to slice whole rat brains (fresh
frozen) and would like the option of slicing whole mice.  (Rats would be
nice, but space and money are limited, which rules out the Leica
Macrotomes.)  Because we work with short-lived radioisotopes, speed is a
very important factor, so I think we need to be able to do both thin slices
and thicker ones (100-300 micron).  We are considering the Hacker-Bright
OTF cryotome.  I don't know if there are other comparable instruments.

I would appreciate any advice on options for freezing and sample handling
as well as cryotome options.  Suggestions for reference reading material
would also be very helpful. The Macro Tape Transfer system looks nice, and
seems to be a larger version of the CryoJane system.  Any feedback from
folks who use either system would be handy.  How useful are they?

I also have some very basic questions:
How long does it take process a rat brain if you are doing 30 micron
slices?
Can you readily switch from 30 micron slices to 100-300 micron slices and
back again?
Can/how should/how long are brain slices stored for later use with receptor
binding assays or autoradiography? (Since one rat brain should give lots of
good 30 micron slices.)
About how long does it take to slice a whole mouse, and what slice
thickness do people use?
Is there a fast and eays way to trim off the areas that aren't interesting?

Thank you very much for your time,
Lynne Jones

Senior Research Technician
Dept. of Radiological Sciences
Washington University Medical School
St. Louis, MO