Re: H & E Staining Method

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Date:Fri, 20 Aug 1999 06:18:18 -0400
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I have two question. What is non-acidified Harris hematoxylin?
Why is it being used for this project? (No sarcasm intended. 
I just don't happen to know about non-acidified hematoxylin.)

Aren't all hematoxylins used in the H&E stain acidified, i.e., 
pH'ed with acetic acid? For our Mayer and Gill, we like the
pH around 2.4-2.5. What is the best pH for Harris hematoxylin?
Would there be a difference in pH if the ripening agent
is mercuric oxide vs. sodium iodate?

Could this be part of the problem with overstaining? Too
high of a pH, making is more non-specific - staining
nucleoplasm and cytoplasm in addition to the chromatin
material? Out of curiosity, are your slides picking up
a lot of blue color too, which would indicate too high
of a pH?

I guess I have more than 2 questions. Oh, well.


Peggy A. Wenk, HTL(ASCP)
William Beaumont Hospital
Royal Oak, MI 48073

Edna_J_Gonzalez/ wrote:
> 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?
> Edna Gonzalez
> PowderJect Vaccines

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