RE: Counterstain for Paraffin embedded sections??

From:"Hagerty, Marjorie A." <>

Methyl green is difficult to work with and if you are not quantifying any
results on an image analyzer requiring a green counterstain, I personally
would not bother with it. 

An option is a pale Hematoxylin. I would recommend;
Dako's Mayer's Hematoxylin,
DAKO Order # S3309.

Preparation of Dako's Mayer's Hematoxylin 

1.	Dilute 1 part of Hematoxylin with 1 part of deionized water.
2.	Filter the solution.
3.	Incubate slides for 2minutes in Hematoxylin
4.	Wash with running deionized water.
5.	Dip 10 times in very dilute Ammonia Water 
6.	Rinse in deionized water 2-5 minutes.
7.	Proceed with dehydration and coverslipping

However, any light hematoxylin will provide sufficient contrast.

Good luck.
-----Original Message-----
From: Kathie Berghorn []
Sent: Thursday, February 08, 2001 2:50 PM
To: HistoNet Server
Subject: Counterstain for Paraffin embedded sections??

	I am hoping someone can help us.   We just recently began 
working with paraffin embedded sections.  We are getting good 
staining with the primary antibodies we are using.  The problem is 
the counterstain.  We would like to use methyl green, but we cannot 
get the tissue to take it up despite our best efforts. We left it on 
for an hour, we made up new methyl green, all to no avail.  We are 
using DAB as the chromogen for our primary antibody staining.  Could 
someone recommend a good counterstain that would provide nice 
contrast and work in paraffin sections?

	Any recommendations would be most appreciated!  Thanks.


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