[Histonet] Eosin-Hematoxylin solution according to Ehrlich(from Sigma)

John A. Kiernan jkiernan <@t> uwo.ca
Wed Oct 26 09:56:26 CDT 2005


Are you sure you have the right name for the solution?

Ehrlich's hematoxylin is an air-oxidized hemalum with a
large excess of alum. The original 1886 formulation is
Water 100 ml
Absolute alcohol 100 ml
Glycerol 100 ml
Glacial acetic acid 10 ml
Hematoxylin 2g
Aluminum potassium sulfate (alum) "to excess" (presumably
saturation).
Leave it for a month before using; it keeps for years.

This is the original recipe, published as a half-page note
in Z. wiss. Mikrosk. 3:150 (1886). It's more easily
available in
"The Collected Papers of Paul Ehrlich", ed. F. Himmelweit,
1956,
Vol. 1 p.113. London: Pergamon Press. I've made and used it
and can vouch for its efficacy as a selective progressive 
nuclear stain. The recipes for Ehrlich's hematoxylin given
in 
books commonly have less alum than the original, and are
consequently somewhat more rapidly acting. 
-- 
-------------------------------
John A. Kiernan
Department of Anatomy and Cell Biology
The University of Western Ontario
London,   Canada   N6A 5C1
   kiernan[AT]uwo.ca
   http://publish.uwo.ca/~jkiernan/
   http://instruct.uwo.ca/anatomy/530/index.htm
_______________________________
Eosin Y is applied as a counterstain following a hemalum
nuclear stain. I don't think you could mix the two together
because eosin Y and related dyes are almost insoluble at 
the low pH of a hemalum solution.
-- 
-------------------------------
John A. Kiernan
Department of Anatomy and Cell Biology
The University of Western Ontario
London,   Canada   N6A 5C1
   kiernan[AT]uwo.ca
   http://publish.uwo.ca/~jkiernan/
   http://instruct.uwo.ca/anatomy/530/index.htm
_______________________________
"Jackson, Johanna" wrote:
> 
> Hi,
> 
> I need to stain frozen rat brain sections (15um) with H&E.  As I'm new to this I bought the ready-made Eosin-Hematoxylin solution according to Ehrlich from Fluka/Sigma.  Does anyone have a protocol for using ready-made H&E solutions?  It seems a bit confusing as I can't do the steps, such as washing etc, that usually go between the haematoxylin and eosin.
> 
> Many thanks,
> 
> Jo
> 
> 
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