Counterstain for Paraffin embedded sections?? -Reply

From:Tony Henwood <>

Check the pH of the Methyl Green (should be around 4). Also keep the
water and alcohol rinses short. Maybe dehydrating in n-butanol might
You could try a haematoxylin counterstain (eg Harris's, Mayers).

Regards, Tony

Tony Henwood JP, BappSc, GradDipSysAnalys, CT(ASC)
Laboratory Manager
The Children's Hospital Westmead,
Locked Bag 4001, Westmead, Australia, 2145.
Tel: (02) 9845 3306
Fax: (02) 9845 3318

>>> Kathie Berghorn <> 9/February/2001 08:49am
	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 counterstains.  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 counterstains that would provide nice 
contrast and work in paraffin sections?

	Any recommendations would be most appreciated!  Thanks.


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