Other histological stains (e.g., methyl green) aside, which would be a better counterstain for nickel-enhanced DAB immunohsitochemistry: cresyl violet or neutral red? Should I alter my staining protocol any or proceed as normal? For example, should I make a conscious effort to stain lighter than normal? Thank you.
For routine IHC we usually combine DAB-IHC with hematoxylin as counter-stain. Brown and light-blue is a nice combination. The nuclear stain is done with diluted hematoxylin to prevent overstaining and masking nuclear antigen-staining.
NI/DAB should form a grey-brown. So red as counterstain, should also provide sufficient contrast.
Cresyl violett and neutral red are "cytoplasma"-dyes. So they may interfere with light antigen-staining in the cytoplasma and mask it.
So, it depends on the localisation of your antigen. DAB usually withstands any counter-staining-procedure you think about. For special questions the counterstain may provide further information about the stained cell, beyond tissue-architecture and nucleus-cytoplasma-relation. In general I think, it would be helpful to stain the sample with and without IHC to see the difference.
Gudrun
Hematoxylin as Gudrun says. For Nickel-DAB, since it is black I sometimes use nuclear fast red (NFR)
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