Re: [Histonet] frozen heart again

From:Gayle Callis

Rena Fail and Vinnie Della Speranza had an article on correcting freezing
artifact on muscle in Histologic XXVI (1), May 2003.  You can go to website and read up on how to correct your problem.  It was
amazing they were able to do this so nicely and simply. 

At 01:39 PM 1/8/2004 -0700, you wrote:
>I have pieces of frozen human heart that was not optomally frozen (meaning
>they just put it in a -70 freezer without cryo protectant OCT or anything).
>I cut off a piece while it was still frozen and mounted some in OCT and made
>frozen sections.  As you can imagine the sections are not good.  The muscle
>is shredded looking with lots of spaces and unrecognizable as muscle.  There
>are cells which could be myocytes but who knows.  I did some IHC on them and
>definately have nuclear envelope staining on some cells.  I am labeling with
>Lap2 and Lamin A&C.  I also tried Ki-67 and CD117-cKit, these were both
>completely negative (good neg. control I guess).  Negatives were negative.
>Is it possible to reserrect this frozen tissue or will it always be ruined?
>If I placed it in formalin and processed it in paraffin what do you suppose
>it would look like.  These antibodies seem hardy and may work on ffpe
>tissue.  These people are doing dna studies on these frozen heart samples
>and are not concerned about preserving the tissue and cell morphology.
>Please provide me with some wisdom here.  Thank you.
>Histonet mailing list
Gayle Callis
Research Histopathology Supervisor
Veterinary Molecular Biology 
Montana State University - Bozeman
PO Box 173610
Bozeman MT 59717-3610
406 994-6367 (lab with voice mail)
406 994-4303 (FAX)

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