[Histonet] Masson's Trichrome Stain

[Camila Gomes]

Dear all,

I did a Masson's Trichrome Stain in frozen liver samples with cancer and it
turns quite well to differentiate the collagen in the tumour from the
normal liver but I would also like to see better the nuclei detail and have
a less intense pink.
Here is the protocol that I am using with the Sigma kit:

(Fixation with 4%PFA for 10 minutes)

1- Bouin's Solution at room temperature over-night;
2- Wash in tap water until remove yellow color;
3- Weigert's Hematoxilin for 5 min;
4- Was in tap water for 5 min;
5- Rinse in deionized water;
6- Biebrich Scarlet-Acid Fucshin  for 5 minutes;
7- Rinse in deionized water;
8- Phosphomolybdic Acid for 5 minutes;
9- Aniline blue solution for 5 minutes;
10-Acetic acid 1% 1'30'';
11-Rinse in water, dehydrate, clear and mount.

All the solution are made at the moment of the staining. I have experience
with the staining but only in paraffin embedded sections and I would like
to know if there is any step to be careful or change with frozen sections.



Anyone can help me please?
Thank you,
Camila