question on pig skin H&E

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From:Gayle Callis <>
Date:Tue, 28 Sep 1999 11:49:35 -0600

What hematoxylin are you using, dipping progressive hematoxylins in
1% HCl/70% alcohol is NOT necessary, in fact can be overkill.

That differentiation step is reserved for regressive hematoxylin (Harris) 
staining, and progressive (Gill I,II,III) staining usually has a very gentle
clarifier, with acetic acid.  You may be removing way too much
hematoxylin with acid alcohol.

Harris hematoxylin with acid alchol step (1 dip is usually enough, and you 
cut the acid to 0.5%) with a good rinse in tap water (what is that pH?)
blue in Scotts tap water for 1 min, rinse for 1 - 2 min before going
to counterstaining portion of H&E.

Gill 1, II, III or Richard Allan Hematoxylin 1, Gill formulations, etc
are progressive, dip in their clarifiers or try 1% acetic.  This removes
background staining with hematoxylin but does not remove much of it,
just don't leave it in for a long time (10 dips was adequate for Gill)
RA hematoxylin 1 took 1 minute in their clarifier, they have directions on
use).  Gary Gill, hopefully, will make further comments - always soooo

Gayle Callis

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