Re: fixed frozen mouse tissues

From:James Mubiru

Dear fellow histonetters
I have limited experience on this issue of processing
frozen tissue. But I did it once and the sections came
out fine. What I did was to get the frozen tissue and
put it NBF to fix and then continued with dehydrating
clearing and embedding. I would like to hear from
those who have had experience with these kind of

James Mubiru

--- "Trenton R. Schoeb"  wrote:
> I've received an unusual request from a PI who wants
> to have HE sections 
> made from some mouse tissues. The mice were perfused
> with paraformaldehyde. 
> The tissues were then treated with 3 changes of 30%
> sucrose buffer, then 
> frozen for cryosectioning. He uses this method for
> studies of the brain. He 
> wants to look at the rest of the mouse to see if
> there are any other 
> manifestations of the mutation he's studying. The
> tissues he has are 
> already prepared like the brains (perfused, soaked
> in sucrose, and frozen). 
> We could do HE stained frozens, but I wonder if we
> can get better fine 
> detail if we paraffin embed the tissues. So, my
> question is, has anyone 
> tried paraffin embedding and sectioning fixed frozen
> tissues such as these, 
> and, if so, was it worth the effort? (I expect we'll
> have to remove the 
> sucrose first.)
> --------------------------------------------------
> Trenton R. Schoeb, DVM, PhD, Diplomate ACVP
> Professor, Department of Genomics and Pathobiology
> University of Alabama at Birmingham
> 205-934-2288
> Department office: 205-934-2117
> Fax: 205-975-4418
> US mail:                    Fed Ex:
> VH 402                      402 Volker Hall
> 1530 3rd Ave. S.            1670 University Blvd.
> Birmingham, AL 35294-0019   Birmingham, AL 35294
> --------------------------------------------------

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