RE: Tissue Adhesion for Immunos

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From:"Todd Sherman" <> (by way of histonet)
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We've tried untreated, Vectabond-coated, and plus-charged slides for our IHC
IP procedures and found that plus-charged slides performed the best when
microwave antigen retrieval was employed.  Sections were mounted in
distilled water only.  Performance was not dependent on whether acidic,
neutral (citrate), or basic buffer solution was used - personal
observations, not scientifically analyzed.  One of the greatest contributing
factors of tissue loss during microwave based HIER (heat induced epitope
retrieval) is the physical action of bubbles in the buffer as the solution
approaches and reaches boiling.

We routinely used microwave HIER in which buffer (including citrate) was
placed in plastic Coplin jars followed by the addition of specimen slides.
The jars (w/ loosened lids) were then placed in a microwaveable container
with distilled water to help distribute heat to diminish zonal heating.
During the heating process, the solution was monitored so that only small
bubbles or no bubbles were allowed.  We were able to retain most of the
section with regularity.  One notable exception was cartilage which would
adhere unless a wrinkle was present in the section during the initial
mounting.  Any wrinkles near cartilage were certain trouble...the typical
result being folding rather than complete loss of the cartilage.

I cannot comment on steaming or pressure-cooking the slides for HIER but I
suspect steaming might be an interesting option for section retention.

Good luck, Todd

Todd Sherman
UT Southwestern Medical Center at Dallas
5323 Harry Hines Blvd.
Dallas, TX  75235

-----Original Message-----
From: Joyce Friedland []
Sent: Thursday, December 02, 1999 2:43 PM
Subject: Tissue Adhesion for Immunos

What methods are all the experts out there using to keep tissue from
loosening, folding over and falling off during Immunoperoxidase
procedures?? The biggest problem is during antigen retrieval using citrate
buffer in the microwave.


*  Joyce Friedland                 *
*         *
*    *

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