[Histonet] Problem with Masson's Trichrome

[Bartlett, Jeanine]

The Gomori's One-Step trichrome is faster than the Masson and should
suit your needs beautifully.

Jeanine Bartlett, HT(ASCP)
Centers for Disease Control and Prevention
Infectious Disease Pathology Activity
1600 Clifton Road, MS/G-32
Atlanta, GA 30333


-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of rschoon
Sent: Friday, August 06, 2004 8:25 AM
To: Bdeer24 <@t> aol.com
Cc: histonet <@t> lists.utsouthwestern.edu
Subject: Re: [Histonet] Problem with Masson's Trichrome


Amber,

Don't know where you got your protocol but I would suggest ANY of the
standerd Histology books.  As for an answer to your problems.

1- mordent in either Boin's fluid or saturated picric acid for 60 min.
at 60oC (there are other time, temp.'s and concentrations if one looks
them up).  The method simply will not work without the modent.

2- use Weigert's Iron Hematoxylin as the nuclear stain.  

Robert Schoonhoven





Bdeer24 <@t> aol.com wrote:

>Hi Everyone!
>
>I was recommended to try the Masson's Trichrome stainings on the slides

>of uterine tissue that I have been collecting (Previously I had only
performed Hematoxylin & Eosin stains). This is a new method to our lab
and thus far, staining has not gone well. Although my sections contain a
significant amount of muscle, I have only been able to see the aniline
blue stain. On a couple of the slides, there was a slight dark purple
coloration on the epithelial lining.
>
>I have noticed upon comparing the protocol we received to others found 
>on the internet that ours lacks the step in which  the slides are
placed in Bouin's fluid. There was also no specification in our protocol
for the type of hematoxylin, so we had been using the same hematoxylin
(Harris's modified with acetic acid) that we use in our H & E stainings.
I was wondering if any of these might have led to the stainings not
going as planned. The tissue sections also looked slightly brittle,
perhaps due to a harsher stain. I am wondering if this has anything to
do with my fixation process in which I use a 4% paraformaldehyde/PBS
mixture, ethanol and xylene steps, as well as embedding the tissues in
paraffin.
>
>Is Bouin's a crucial step, would I have better staining with a 
>different tissue fixation, or should I be looking in a different 
>direction? Any suggestions would be very much appreciated! Thanks!
>
>Amber Greenbank
>
>Arizona State University
>
>_______________________________________________
>Histonet mailing list
>Histonet <@t> lists.utsouthwestern.edu
>http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>  
>



_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet