[Histonet] sections falling off slides

[Rene J Buesa]

Your test subject is one of the more difficult there are.
Although you may get some background, use some gelatin dissolved in the water bath to try to adhere the sections to the slides. Also try to cut as thin as you can and make sure there is no water left between the sections and the slide.
Bake at 60ºC for at least 45 minutes before starting the IHC procedure. If you use HIER try to switch to pepsin.
The least "stress" you add to the sections the better.
René J.

From: Robin Dean <robin_dean <@t> compbio.com>
To: histonet <@t> lists.utsouthwestern.edu
Sent: Monday, May 16, 2011 2:07 PM
Subject: [Histonet] sections falling off slides

Hi All,



We are trying to do IHC for multiple antigens on goat knees. The sections
are decalcified FFPE sections and we are using a Dako autostainer. We are
currently trying air-drying for  1-2 days  and baking of tissue slides
before use in IHC..

The one previous time we tried baking, we noticed a decrease in IHC signal.
The sections are pretty large, we tried cutting them down, but can't
decrease the size much.  We also  decreased the number of washes and size of
washes on the Daok, which helped only slightly.  Does anyone have any
suggestions?



Thank you in advance.



Robin



Robin R. Dean, Ph.D.

Senior Scientist & Study Director

Comparative Biosciences, Inc.

786 Lucerne Dr.

Sunnyvale, CA

(408) 738-8060

robin_dean <@t> compbio.com



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