RE: uneven IHC staining patterns

From:"Dawson, Glen"


I experienced the same thing and was dumbfounded for a while.  In a last
ditch effort, I tried adding some additional tween (soap) to my wash buffer
to help break down surface tension and facilitate even spreading of reagent
across the whole slide because I observed a tendancy for some reagent to
pool up in various areas on the slide.  I only added 1ml of tween per 10 L.
of wash buffer but it seems to have helped.

Best of Luck,

Glen Dawson  BS, HT & QIHC (ASCP)
Lead IHC Technologist
Milwaukee, WI

-----Original Message-----
From: Pat Zeitlow []
Sent: Tuesday, September 10, 2002 11:51 AM
Subject: uneven IHC staining patterns

We are periodically getting uneven staining patterns on our IHC slides.
I have exhausted my troubleshooting skills and need input from anyone
out there who may be able to help.  We are using the Dako Autostainer (2
different ones).  I have checked the leveling, reagent distribution, and
slide position.  I am in a reference laboratory and unable to do much in
terms of trouble shooting fixation, etc.   I am at wit's end and will
happily provide any other details to assist.  Dako says they are
clueless and have heard no other similar complaints.  Please help.  TIA.
Pat Zeitlow
Molecular Pathology Manager
Boyce and Bynum Laboratories

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