[Histonet] Re: Substrates

[Johnson, Teri]

Margaret,

This has been discussion on the Histonet before, and while I have not
personally done this, I've heard it works:

>>Try counterstaining the immuno slides with giemsa rather than
performing the
bleaching technique.  I use the May Grunwald giemsa following DAB.  The
melanin
pigment will stain green and contrasts nicely beside the brown DAB.

Dr. H Kamino originally published counterstaining with azure B to color
melanin.
I learned this trick from her when she was the chief of dermatopathology
here at
Duke.

We routinely use this combination.  One note of interest, sometimes the
DAB
color reaction is enhanced or altered because of prior treatment of the
tissue,
such as antigen retrieval or enzyme digestion.

Jim Burchette
Duke Immunopathology<<

You might also consider using Vector's Nova Red which works well,
provides nice contrast with hematoxylin, and is stable to alcohol and
xylene. That way you would only have to change the chromogen and nothing
else in your protocol.

Teri Johnson, HT(ASCP)QIHC
Managing Director Histology Facility
Stowers Institute for Medical Research
1000 E. 50th St.
Kansas City, MO 64110