We had that same problem last year.  We tried everything!!!  It was very
humid in our lab and we thought that might be the problem.  We tried using
molecular sieves in our last two Pro-Par stations on the stainer.  That
helped a little but we still had the problem.  Finally, we switched to a
different mounting medium.  After trying many brands, we found that CMS
protocol (Xylene) works great and we no longer have the problem.  (We were
using Refrax).

Michelle Fuller
ACM Medical Laboratory
Rochester, N.Y.


-----Original Message-----
From:	Hess, Mary [mailto:mary.hess@canji.com]
Sent:	Monday, September 25, 2000 5:24 PM
To:	'histonet@pathology.swmed.edu'
Subject:	Xylene substitutes

I've received some responses asking why we used xylene for Masson's
trichrome instead of pro-par in the first place. Initially it was because I
got the Masson's Trichrome protocol and the kit from another person at my
company who was very much into using only 'pure' reagents (ie, expensive
absolute alcohols, xylene, etc) to ensure the quality of his slides. And the
quality is very good when you use those solutions. When I started, I quickly
switched to our normal alcohol and found no difference between his slides
and mine. I have now tried to do our Masson's trichrome using pro-par and
refrax mount instead fo xylene and permount. My initial results aren't too
good.
Recently we have found eosin 'bleeding' from our H&E tissues when the slides
were mounted (for which we used Pro-Par and Refrax mounting media). I
switched out everything (from water, to alcohol, to Pro-Par, to eosin and
hematoxylin) in our H&E stations and still had the problem. When I did the
trichrome with the Pro-par,  the Biebrich Scarlet Acid Fuscin bled into the
mounting media. I don't know what it is. If you guys do I would be grateful.
And in the end, it was the guy who started me on the stain who kept me using
xylene. I don't want to keep using it, but so far, it seems like I'm going
to have to. I hope this 'bleeding' is something I'm doing wrong and is
easily fixable, I'd rather it be my fault. Thanks for the help so far....
-Mary Hess, San Diego, Ca.