[Histonet] Does anyone know how to achieve a perfect Masson Trichrome stain. Our counterstain (light green in acetic acid) works well but the intensity of the muscle stain is not very bright. Does anyone have any suggestions? We will be staining liver

[Lee & Peggy Wenk]

Lots of questions, as this is one of those stains that has a lot of places
that need to be tweaked:

NEW OR OLD? Is this a new problem you are having, or have you always had
this problem?

MORDANT: Are you still post-mordanting in Bouin, or have you tried to
eliminate this or switch over to something else to try to not use picric
acid in the lab? What time and temperature?

RED DYE: What dye(s) are you using for the red dye for the muscle? Please
include name and color index (CI) number, if that's available in the UK.
Also, have you switched recently to a new type of red dye, or purchased a
replacement when you ran out? What is the dye content percent on the dry
powder?

PMA/PTA: Are you using Phosphotungstic acid and/or Phospomolybdic acid? How
old is it?

ONE OR TWO STEP: Just to make certain, is this a true Masson trichrome,
where the red dye is applied, and then the PTA/PMA, and then finally the
green dye? Or is this the traditional Gomori, where the red and green dyes
and the PTA/PMA are all in one solution?

TIME IN RED AND GREEN DYES: Are you trying to standardize the time in the
red dye and the time in the green dye? This usually doesn't work. Each
different type of tissue, and each different different person's tissue, will
pick up red dye and green dye at a different rate. 

TEMPERATURE: Are the dyes at room temp or refrigerator temperature? That can
have an influence.

MACHINE OR HAND: Are you hand staining, where you have control over the
amount of time in each solution, where you can check the progress with a
microscope, where you have a choice of dyes? Or are you staining with a kit?
Or on a machine?

Sorry, need some more information, before we can help you pinpoint the
problem.

Peggy A. Wenk, HTL(ASCP)SLS
Beaumont Hospital
Royal Oak, MI 48073

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of SARAH REEVES
Sent: Tuesday, December 30, 2008 10:09 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Does anyone know how to achieve a perfect Masson
Trichrome stain. Our counterstain (light green in acetic acid) works well
but the intensity of the muscle stain is not very bright. Does anyone have
any suggestions? We will be staining liver bio

Does anyone know how to achieve a perfect Masson Trichrome stain. Our
counterstain (light green in acetic acid) works well but the intensity of
the muscle stain is not very bright. Does anyone have any suggestions? We
will be staining liver biopsies mostly in the near future and wish to
improve our technique.

Thanks in advance

Sarah


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