[Histonet] Masson's trichrome

[Bryan Llewellyn]

Go to http://stainsfile.info and read up the principles on which trichrome 
stains work.  I suggest you then start with the standard Masson method 
listed there.  It is one of the easiest to control.  The commonest reason 
for getting all green (or blue) staining is the fixation.  Formalin fixation 
is the likely cause.  Either modify the staining protocol to 10-2-2 minutes 
with plasma-polyacid-fibre stains, or refix sections in Bouin's fluid for an 
hour at 56C.  The nuclei are not usually black, regardless of what the books 
say they should be.  They are usually a muddy brown.

Bryan Llewellyn


----- Original Message ----- 
From: "Julien LambreyDeSouza" <julien_lambreydesouza <@t> uqar.qc.ca>
To: <histonet <@t> lists.utsouthwestern.edu>
Sent: Wednesday, May 04, 2005 10:30 AM
Subject: [Histonet] Masson's trichrome



Hello,

I am trying out Masson's trichrome for the first time. I used the Goldner
modification from Gabe 1976, with Fuschine-Ponceau, orang-G phosphomolybdic
and Light green. We are staining 7 micron sections of entire fish larvae
heads. When following the protocol (5 minutes in each color followed by a
1% acetic water rinse) I get a general green staining of all tissues. The
only way to differentiate cartilage from the rest in our section is by
recognizing cellular topography, same as with Hematox-eosine staining,
therefore loosing the whole point in using Masson's trichrome. I seem to be
doing something wrong.

Would anybody familiar with the technique be inclined to share their
expertise or protocol? I would be happy to send a pic of our staining
result by email to have input on the possible problem.

Thanks,

Julien.
Julien Lambrey de Souza
Assistant de recherche, Biologiste M.Sc.
Biologie Évolutive.
Université du Québec à Rimouski
Département de Biologie, Chimie et Sciences santé
300, allée des Ursulines
Rimouski (Québec) Canada G5L 3A1
Tél.: (418) 723-1986 ext. 1714
Fax.: (418) 724-1849
Courriel: julien_lambreydesouza <@t> uqar.qc.ca

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