Re: H & E Staining Method

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From:"Tony Henwood" <>
To:"HistoNet Server" <>, Edna_J_Gonzalez/
Date:Sun, 29 Aug 1999 20:15:46 +0000
Content-Type:text/plain; charset=US-ASCII

Dear Edna,
> I am working with pig skin and doing H&E regressive staining using Harris
> Hematoxylin (non-acidified). The problem is that the hematoxylin is too
> dark and I can't differentiate what I need, especially it is too dark in
> the basal layer and the epidermis of the skin. I need a H&E Regressive
> Method that will be lighter. Currently I am staining with hematoxylin for
> 10 minutes, which I think is too much, but this is what one of the methods
> say. I know there are different H&E regressive methods (with different
> times for Hematoxylin). Any suggestions?

Keratohyaline granules, the basel layer is rich in them, will stain 
strongly with haematoxylin. Try shorter staining time but possibly 
the problem may lie in the thickness of the sections. Thinner 
sections will decrease the concentration of these granules available 
for staining. A useful QC is to look for capillaries. Each cross 
section should have 1-2 endothelial nuclei present. Any more and the 
section could be too thick.

I hope this helps.

Regards, Tony.
Tony Henwood
Senior Scientist
Anatomical Pathology
Royal Prince Alfred Hospital

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