[Histonet] DAB haematoxylin counterstain; too purple, overpowering IHC

[Megan French]

Hi all,

 

I have been working through an immuno using DAB and counterstaining with
harris haematoxylin. 

 

Protocol for counterstain:

 

5 min h20

1 dip haematoxylin

1 min h20

2 dips Blue in ammonia

1 min h20

Dehydrate, clear, mount

 

I am finding that my slides are coming out realllly purple and are
overpowering my DAB staining so much so that I can't even see it! 

The haematoxylin is new, it was recently changed. I don't think it has
anything to do with the DAB/quenching etc im pretty sure is is haem
related.

 

Any suggestions would be appreciated!!

 

 

Megan French 

Surgical Research; Murdoch Childrens Research Institute

E megan.french <@t> mcri.edu.au


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