[Histonet] substrates

[Susan Q Wells]

You may lose your epitopes with this procedure, if so 10% H202 in PBS 
for 30 minutes may lighten the melanin pigment enough
to view  your chromagen. I  recently used Vulcan Fast Red and Bajoran 
Purple from Biocare Medical with great success where melanin pigment was 
present.

Sue Wells HT(ASCP), QIHC

Rene J Buesa wrote:

>Margaret:
>  Confronted with that situation I would eliminate the melanine better. Oxidize it with 0.1% potassium permanganate for 2 hours; wash and remove the oxidized melanine with 0.3% oxalic acid followed by washing. The section should be free of melanine pigment/colour afterwards and you can use DAB (for brown) and DAB + Ni salts for dark blue as double substrates.
>  Hope this will help you!
>  René J.
>
>"Perry, Margaret" <Margaret.Perry <@t> sdstate.edu> wrote:
>  We are trying to do double staining and are looking for two different
>colors of substrate and we can't use DAB due to melanin. Does anyone
>know of a green stain that is xylene compatible? Vector has Nova Red
>and a blue color but we use hematoxylin as a counterstain. It would
>mean an extra step for us to counterstain in Methyl Green
>
>We use a DAKO autostainer.
>
>
>
>Margaret Perry HT(ASCP)
>
>South Dakota State University
>
>Animal Research and Diagnostic Lab
>
>Brookings SD 57007
>
>Margaret.Perry <@t> sdstate.edu
>
>
>
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