[Histonet] Antigen retrieval and rat skin

[Yaskovich, Ruth A (NIH/NIDCR)]

Ron,
I just did mine they are formalin fixed paraffin sections. They stayed on
during the retrieval process just fine. I only bake for 20 mins. and then
let them get to room temperature. Just put a drop of formalin in the bottom
of dish or jar.
Hope this helps.
Ruth Yaskovich

> ----------
> From: 	Ronald P. Wilson
> Sent: 	Thursday, June 3, 2004 12:18 PM
> To: 	Histonet
> Subject: 	[Histonet] Antigen retrieval and rat skin
> 
> Thank you for all the replies so far. As for additional details about our
> procedure that some requested, the tissue is formalin-fixed and
> paraffin-embedded. Sections are cut at 4 microns, mounted on Superfrost
> Plus
> slides, air-dried and baked in a 55-60ºC oven for 1 hour before starting
> the
> procedure. We use a Black and Decker rice steamer in which we place 3-4
> plastic coplin jars containing citrate buffer (pH 10) into which the
> slides
> are loaded back-to-back and standing upright. We steam for 20 min; the
> steamer is started ahead of time so that the temperature reaches 96-98ºC.
> The slides are then allowed to cool for 20 min before continuing with the
> staining procedure. We recently tried some capillary gap slides and used
> the
> reagent reservoirs in place of the coplin jars. There seemed to be a
> slight
> improvement in the resulting slides but still had some disruption of the
> dermis.
> 
> One reply recommended covering the tissue with about 1/2 ml of 10%
> formalin
> and then placing in a 60ºC oven. Is this for frozen sections or
> formalin-fixed paraffin-embed sections? I am willing to give it a try. We
> believe that the problem is a result of the antigen retrieval with heating
> since our standard H&E sections and BrdU sections which uses 20 min
> trypsin
> digestion have very minimal wrinkling or disruption of the dermis.
> 
> Again, any suggestions or comments are much appreciated.
> 
> Ronald P. Wilson
> Department of Comparative Medicine, HO54
> Penn State University College of Medicine
> M. S. Hershey Medical Center
> 500 University Drive
> Hershey, PA 17033
> phone:  717-531-8460
> fax:       717-531-5001
> e-mail:  rpw4 <@t> psu.edu
> 
> 
> 
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