RE: intern seeks help...

From:Bert Dotson <> (by way of histonet)

All trichromes are extremely fixation dependent. For this reason, post
fixation of hydrated slides prior to staining is usually recommended. The
usual treatment is 30 minutes to an hour in Bouins solution at 60 degrees.
If you are using frozen (instead of paraffin processed) tissue sections,
you will probably need a procedure specifically drawn up for frozen


-----Original Message-----
From:	Heather Hall []
Sent:	Wednesday, September 13, 2000 12:20 PM
Subject:	intern seeks help...

My name is Heather Hall and I am presently doing an
internship at Primedica Inc., located in Worcester,
Mass.  One of the required slides is skin stained with
trichrome.  For some reason, I cannot get this slide
to come out right.  I first tried Masson's procedure,
with kit reagents.  The slides appeared as though they
had not been differentiated in
phosphotungstic/phosphomolybdic acid long enough, as
the collagen bundles were retaining the scarlet-acid
fuchsin in their centers.  My first step to correct
the problem was to increase the time in the p/p acid;
that didn't work.  Next I made up fresh reagents;
didn't work.  Altering the times in the other reagents
didn't work either, so I tried the Gomori's procedure.
 When the Gomori's did not work, I changed species
(first species used was pig, second was human).  The
same problem was occuring in the human tissue also.
Since the control slides appeared to coming out fine I
tried a third species.  I tried rat skin hoping that
it might have a thinner connective tissue layer and
permeability would be altered, but that came out as
badly as the others.  I don't know what to do next to
try and fix this problem.  Any suggestions would be
very much appreciated.  Thanks for any help you may be
able to give.
Heather Hall
Primedica Inc.
Histology Lab
Worcester, MA

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