[Histonet] RE: Counterstain to use with blue and red substrates

[C.M. van der Loos]

Dear Douglas,
I presume that the Methyl green that has "covered some of the blue" as you said, was due to the fact that it diffuses away after aqueous mounting needed after using AEC as chromogen. You may try to replace AEC by Vector NovaRed or Romulin Red (BioCare), two peroxidase chromogens that stand organic mounting using VectaMount (a non-xylene based organic mountant) very well. Although the red color is more brownish than with AEC it still contrasts nicely with the dark-blue NBT/BCIP reaction product. Perhaps now your Methyl green counterstain will work for you.
Lots of success and let us know! 

Chris van der Loos, PhD
Dept. of Pathology
Academic Medical Center
Amsterdam - The Netherlands


----- Original Message ----- 
>From  d.gregg <@t> juno.com 
Date  Wed, 26 May 2004 13:03:19 -0400 
To  histonet <@t> lists.utsouthwestern.edu 
Subject  [Histonet] Counterstain to use with blue and red substrates 
Hi all,
I am using blue (NBT) and  red (AEC)  substrates in a double stain and
wondered if anyone has a recommendation of a nuclear counterstain that
may work with this. I could use metanil yellow a a cytoplasmic stain but
would prefer a primarily nuclear stain. I have used Methyl green but it
has covered some of the blue in the past. Thanks.

Douglas Gregg
Plum Island Animal Disease Center 
Greenport, NY