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

[Due, Brice]

The Gomori used in muscle panels is the ENGEL-CUNNINGHAM MODIFICATION which
adjusts the pH of the standard Gomori one-step. As far as I know it only works
properly on fresh unfixed frozen sections. It is very sensitive to the condition
of the tissue (fixation, lipid extraction, freeze quality, fresh bx drying,
etc.). Online, see:
http://www.neuro.wustl.edu/neuromuscular/pathol/histol/trichrom.htm

-brice

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-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu]On Behalf Of John
Kiernan
Sent: Friday, August 12, 2005 4:11 PM
To: Hofecker, Jennifer L
Cc: histonet
Subject: Re: [Histonet] problems with Gomori's Trichrome on Frozen
Muscles


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
   kiernan[AT]uwo.ca
   http://publish.uwo.ca/~jkiernan/
   http://instruct.uwo.ca/anatomy/530/index.htm
_______________________________
"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
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
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