Re: [Histonet] problems with Gomori's Trichrome on Frozen Muscles

From:John Kiernan

If the method is Gomori's one-step trichrome, with
chromotrope 2R and light green (or fast green FCF),
and PTA or PMA, then muscle should stain red, and 
collagen green. Instructions for 2 variants of the 
method are given in Presnell & Schreibman (1997) 
Humason's Animal Tissue Techniques, 5th edn. 
Baltimore: Johns Hopkins Univ. Press, pp.129-131.
I think it's also in earlier editions of the
book (with G. Humason as the author). And in 
other books too, of course.

Trichrome methods are usually done on paraffin 
rather than frozen sections. If the fixative
is formaldehyde, staining is improved by a
pre-treatment of the hydrated sections with
picric acid - Bouin's solution is often used.
John A. Kiernan
Department of Anatomy and Cell Biology
The University of Western Ontario
London,   Canada   N6A 5C1
"Hofecker, Jennifer L" wrote:
> Happy Friday everyone,
> We are experiencing a strange phenomenon with our Gomori's.  All of a sudden the staining is predominately red (instead of green).  Normal muscle control is still green but the patient tissues are staining red.   We have tried several different changes: new trichrome solution, recutting frozens, etc.  The only thing that is reproducible is that sections which have been previously cut and stored at -70 seem to stain fine.  If we cut a frozen then begin staining it without time in the freezer, voila, the same muscle sections are now red instead of green.  What could be causing this?  I even pulled out the same piece of control tissue and cut fresh slides on it.  They were red also.  There was one fresh slide that stained green in the whole "experiment" otherwise, anything that doesn't spend time in -70 is turning red.  We are using the same procedure that we have been using for quite a while, and the precut slides do stain, so I don't think it's a procedural error.   The only change
> is that we have been trying to stain the trichromes when the specimens are cut instead of storing in the freezer to batch staining later.  Has anybody experienced this?  Any ideas would be appreciated.
> Thanks in advance,
> Jennifer
> Jennifer Hofecker, HT (ASCP)
> Vanderbilt University Medical Center
> Division of Neuropathology
> (615) 343-0083
> (615) 343-7089 fax
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