Re: [Histonet] Washed out spots

From:"Gudrun Lang"

Maybe the deparaffination-step before staining is insufficient. On spots of
residual paraffin the hematoxylin will not work well.
We deparaffinise in two baths of xylen-substitute for at least 10 minutes in
Gudrun Lang

----- Original Message -----
From: "Fran Lemons" 
Sent: Thursday, March 18, 2004 3:13 PM
Subject: [Histonet] Washed out spots

Hello fellow 'netters!
I am trying to troubleshoot a problem with some of our sections, perhaps you
can give me some input.
Some of our sections, mainly the biopsies, have areas of "washed out" stain.
That is to say, there are "spots" where the tissue just didn't seem to take
up the hematoxylin.  We use the Gemini Varistain autostainer, and the
reagents are fresh & in order as they should be.  Our biopsies are processed
on a 4 hour run separate from large pieces, and all of the reagents on the
processor (VIP) are as they should be.  I even considered perhaps something
in the waterbath that is getting on the sections and "blocking" the
hematoxylin, such as a cleaning agent residual; but I am assured by my techs
that they use Paragard on the microtomes and hot water only on the
waterbaths.  Biopsies are cut before the regular run, maybe there is
residual on the 'tomes?  Dunno.
Didn't mean to write a book, but I wanted to provide as much info as
possible in case someone else out there is experiencing the same problems.
Any suggestions would be helpful.
Thanks in advance,
Fran Walker
BHET Knoxville

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