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

[Bonnie Whitaker]

Jennifer,

If I understand correctly, you are probably doing frozen muscle biopsies for
a neuropathologist as part of a whole muscle biopsy work-up.  The Gomori's
modification used by neuropathologists does indeed produce green muscle, if
the muscle is normal.  Only "ragged red fibers" should be staining red.  I
really don't know the specifics of this, but you don't do the bouins
post-fixation.  As I recall a friend and former co-worker had pH problems
that caused some variances in her muscles w/ Gomori's.  I have some muscle
experience, but am not currently doing that, and don't have access to the
references I used.  

Maybe someone with expertise in neuropath will address this issue with more
clarity than I can.

Bonnie Whitaker


-----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 3:11 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
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet

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