[Histonet] Modified Gomori Trichrome

[Marie Wattenhofer]

Hi
I am a beginner in histology....
I would like to do the modified gomori trichrome staining to see if I have
RRF on muscle frozen sections.

I tried the following protocol that I found on the web

Preparation of solutions:

SolutionA:
Chromotrope 2R 60 mg
FCF 30 mg
phosphotunstic acid 60 mg
glacial acid acetic 1 ml
Water up to 100ml
Adjust pH to 3.4 with  HCL 37%

Solution B
200 ul of glacial acetic acid in 100 ml of water

Protocole
Frozen muscle section
3 mn in filtred Hematoxiline Harris
Wash 3 times in water
10 mn in Solution A
Wash 3 times in water
10 seconds in solution B
Deshydratation: EtOH 70%, 95% absolute
Histosol 10 mn

I got the following result.
>From the border to the center of the cut the coloration was not the same:
At the border of the cut, fibers are green (great!) but then they are red
(???) and at the very center, they are not colored at all.

Could someone tell me what I did wrong?
should I stain longer in solution A ?
I have read that the pH of solution A is VERY important; was it ok to pH
with HCl?
Is the solution B formula correct?

Thanks for your help!

Marie



-- 
Marie WATTENHOFER-DONZE
Institut de Genetique et de Biologie Moleculaire et Cellulaire (IGBMC)
1 rue Laurent Fries, BP 10142
67404 Illkirch Cedex
France
Tel. 33+(0)3 88 65 34 16
web site : http://www-igbmc.u-strasbg.fr