RE: Thawing frozen tissue for H&E
|From:||"Monson, Frederick C." |
While this is NOT a suggestion from a Pathologist, it may help by
giving you a position to start from.
I have, in the past, used what colleagues called a smart-a__
solution to the same problem. I had frozen tissue and wanted H&E permanent
slides. The key is to provide the tissue in a 'recognizable' form/condition
to the H&E even though it arrives with much more baggage that is normally
lost in fixing, dehydrating and cooking. What I did was as follows.
OCT section on slide.
With section confined to middle 1cm of slide, I filled the bottom of
a Coplin jar (screw cap) [anything appropriate will do] with 1cm of whatever
HCHO fixative I had on hand (I used 4% pHCHO [HCHO from paraformaldehyde] in
I carefully lowered the slide into the Coplin Jar directly from the
cryostat and screwed on the top. I placed the jar in the 'fridge'
Next morning, I removed the slide and ran it through the H&E
sequence. Staining was shorter and differentiation was longer and eosin
concentration was lower by a quarter, but I finally got it right.
Then I ran the slide up to xylene and mounted with my favorite, Gum
Damar [home made]. All of the above was true as long as the section stayed
where I put it after I cut it!
This method worked most of the time. Please note, everything I did
was in one's or 5's or 20's, and nothing was done automatically after tissue
was infiltrated in my Technicon - but that's not related to this.
Frederick C. Monson, PhD
Center for Advanced Scientific Imaging
Schmucker II Science Center
West Chester University
South Church Street and Rosedale
West Chester, Pennsylvania, USA, 19383
CASI URL: http://darwin.wcupa.edu/casi/
WCUPA URL: http://www.wcupa.edu/
Visitors URL: http://www.wcupa.edu/_visitors/
> From: Richard Cartun
> Sent: Friday, June 21, 2002 2:33 PM
> To: Histonet@pathology.swmed.edu
> Subject: Thawing frozen tissue for H&E
> We received 3 biopsies of colon to R/O Hirschsprung's disease.
> Unfortunately, all of the tissue was frozen in O.C.T. so we have nothing
> for permanent sections. What is the best way to thaw the frozen tissue
> before submitting it for H&E? Should we trim away the excess O.C.T. and
> place it in formalin or should we "wash" it in tissue culture media
> (RPMI) before placing it in formalin? Obviously, preservation of
> morphology is critical here. Thanks!
> R. Cartun
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