Re: [Histonet] Melanin, fast red, and a counterstain
Red chromogens are excellent for giving a red/blue contrast with
hematoxylin, and having the melanin pigment there too, you will have three
Our researchers prefer red chromogens with hematoxylin, and we never use
methyl green with red, but have used it with enhanced DAB (black
color). Most of our researchers are used to seeing hematoxylin stained
sections and this counterstain is helpful for additional morphology
examination. Be sure to not be heavy handed with the hematoxylin, you
want to LIGHTLY counterstain so you don't mask your red immunostained cells.
You can always counterstain one slide with hematoxylin and not counterstain
an adjacent section just to see the different color contrasts, you may
actually prefer NO counterstaining.
We never use methyl green with red chromogens, but have used it with brown
DAB or an enhanced black DAB.
Be careful, methyl green may leach out of the section with a red AEC when
you coverslip with aqueous mounting media as AEC will wash out/fade in
alcohol or clearing agents. Vector NOVA red is permanent and will not fade
after exposure to solvents. So be aware of mounting media compatibility
with the chosen chromogen.
At 11:07 AM 10/30/2006, you wrote:
>Thanks to everyone who answered the questions about melanin pigment. It
>seems most of you use a red pigmented chromogen to demonstrate IHC
>staining that will be easy to differentiate from melanin. I should have
>asked this question at the same time: do the red chromogens work all right
>with a hematoxylin counterstain, with a methyl green counterstain, or can
>I do without the counterstain entirely? Thanks again.
>Melville B. Vaughan, Ph. D.
>Department of Biology
>University of Central Oklahoma
>100 N. University Drive
>Edmond, OK 73034
>Histonet mailing list
Research Histopathology Supervisor
Veterinary Molecular Biology
Montana State University - Bozeman
PO Box 173610
Bozeman MT 59717-3610
406 994-4303 (FAX)
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