Re: [Histonet] substrates

From:"Katri Tuomala"

Margaret & Rene,

Just a caution: the bleaching the melanin before immuno stains may abolish 
or diminish some antigen/antibody reactions.

Katri

Katri Tuomala
Hamilton, Ontario, Canada

----- Original Message ----- 
From: "Rene J Buesa" 
To: "Perry, Margaret" ; 

Sent: Wednesday, June 14, 2006 10:23 AM
Subject: Re: [Histonet] substrates


> 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"  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@sdstate.edu
>
>
>
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