Suggestions:
1) If it's compatible with the autostainer that you're using, add a
surfactant such as Tween 20 (0.05%) to your buffer rinse solution, in order
to cut the surface tension and allow the staining reagents to flow across
your slides more evenly.
2) If it's possible, add more volume of dispensed reagents to each slide.
3) If it won't affect your positive staining intensity, decrease the
incubation times in reagents.
4) Make sure there isn't an excess of air flow across your slides (gap in
autostainer cover seal).
5) Make sure your autostainer is level.
6) As others have mentioned, make sure that your slides are completely
dewaxed.
Jan Shivers
UMN VDL
----- Original Message -----
From: "Paula Lucas" <plu...@biopath.org>
To: <histonet@lists.utsouthwestern.edu>
Sent: Wednesday, January 21, 2009 6:04 PM
Subject: [Histonet] Auto-IHC Staining Issues
Hello,
I've been having a problem with the staining quality from an auto-IHC
stainer and I was wondering if any of you experience the same thing.
I notice sometimes that the staining in the tissue isn't complete, like
there are areas where the fluid (one or more of the steps involved) didn't
get in contact with the tissue, thus resulting in patchy, skipped staining.
I'm thinking that this must be a dispensing problem.
Is this a general thing with all auto stainers - ones like DAKO or Lab
Vision's stainer or Biocare Medical's stainer? These stainers dispense
fluid onto the slide.
I hope this explains things and I really hope that I can get some feedback.
It's getting very frustrating.
Thanks in advance,
Paula Lucas
Lab Manager
Bio-Path Medical Group
Fountain Valley, CA
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