[Histonet] mouse skeletal muscle frozen section prep

[Merced M Leiker]

I've frozen numerous rodent (including mouse) skeletal muscle tissue for 
sectioning. I've never heard of using talc before. I just blot any moisture 
off the tissue with a paper towel or Kimwipe. I usually do not fix the 
tissue beforehand but after sectioning, on the slide. To freeze, I just 
immerse the tissue into a cryomold filled with OCT and freeze over liquid 
nitrogen vapors or, since you don't have isopentane, in a slurry of dry 
ice/isopropanol (don't let the alcohol spill over into the freezing OCT, 
however, or you'll have to scrape it off as alcohol won't freeze). Or you 
can slowly dip the mold into some liquid nitrogen. I wouldn't just throw it 
in, however, as I used to do that and ended up with half my OCT blocks with 
cracks in them, right through the tissue.

Hope this helps! I'm sure others on Histonet have frozen mouse skeletal 
muscle as well.

Regards,
Merced

--On Tuesday, February 08, 2011 10:51 AM -0500 "Spinette, Sarah" 
<sspinette <@t> ric.edu> wrote:

> I am new to working with mouse skeletal muscle and have been reading
> about various methods for preparing and freezing tissue for sectioning.
>
> I have heard of the benefits of coating the tissue in talc prior to
> freezing but have not yet seen a single protocol that describes the use
> of talc and OCT or other medium IN plastic molds, can you still use talc
> and then plunge the tissue into cold OCT in the mold end then plunge the
> entire block into LiN2 (we do not have isopentane)?
>
> Also, can anyone comment about when and how they choose to fix the tissue
> first and if they use paraformaldehyde before freezing?
>
> Thank you!
>
>
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Merced M Leiker
Research Technician III
Cardiovascular Medicine
348 Biomedical Research Building
State University of New York at Buffalo
3435 Main St, Buffalo, NY 14214  USA
leiker <@t> buffalo.edu
716-829-6118 (Ph)
716-829-2665 (Fx)

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