[Histonet] Methyl green going away...

[Goins, Tresa]

I don't know if this will apply to your application, but after the chromogen and counterstaining steps, we rinse with distilled water and air dry our IHC slides (oven at 60C for 30 min) rather than dehydration through alcohol and xylene.  Works great with permanent red and hematoxylin.

Good luck,

Tresa

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Alonso Martinez-Canabal
Sent: Monday, April 16, 2012 9:27 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Methyl green going away...

  Hi 

my boss requires me to counterstain some DAB immunohistochemistry with methyl green. I use for that purpose the methyl  green in aquos solution 0.5% in 4.2 acetate buffer. The problem is that after the washes I dehydrate and the methyl green becomes extremely faint with the alcohols. My protocol requires long periods of alcoholic dehydration very graded. Is there any idea on how to save the methyl green. Can I use an alcoholic solution?
Thank you very much. 

Alonso 

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