RE: [Histonet] IHC ~ Tissue Falling Off Slides

From:"Jeff Gordon"

Laura, what drying technique are you using?  While the problem may possibly be the slides, often we are able to solve tissue adhesion problems with proper slide drying.  We use standard positively charged slides with no special coating and no special adhesive in the water bath.  We pretreat slides in an electric pressure cooker for 15 minutes with Trilogy EDTA, which is a pretty aggressive pretreatment, and we never have recurring problems with tissue adhesion.  Our slide drying technique is 2 hours at 60 degrees Centigrade for most tissues (super sensitive tissues like fatty breast we will dry for 4 hours at 60 degrees), and the tissue stays on just fine in the pressure cooker pretreatment.

Jeff Gordon
Cell Marque Corp.

-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Jones,
Laura
Sent: Thursday, December 09, 2004 10:04 AM
To: Histonet (E-mail)
Subject: [Histonet] IHC ~ Tissue Falling Off Slides


Hello fellow Histonetters.  

We have suddenly started to have problems with tissue falling off slides
during IHC processing.  We suspect the problem may be our slides, although
we have been using the same ones from Mercedes Medical for a while and they
have worked beautifully!  We have contacted them, and they are investigating
and very kindly replacing our stock, but in the meantime, our Pathologist
has asked me to ask all of you where else we can find 25 X 75 silinated
slides.  We use the Zymed ST-5050 automated stainer, and the 1 X 3 slides
are sometimes a bit to big to fit on the carousel.  

Just for additional info, we seem to be experiencing problems mostly with
the more alkaline retreival solutions - AR-10 (Biogenex) and EDTA (Zymed)
and Trilogy (Cell Marque).  We use the Black & Decker steamer, and have
adjusted our times in retrieval from 75 minutes (!) to 20 minutes, per
Pathologist direction.  We have also tried 20 minutes heating, then adding
the slides for 20 minutes of boiling, then allowing them to stand for 20-30
minutes.  

Thanks in advance for all of your knowledge!  

The Histochicks at Sharon Regional

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