I am using ICC to stain some nuclear proteins in muscle-derived stem cells. I'm currently using a HRP-conjugated secondary and DAB substrate (From Biogenex) I would like to avoid haematoxylin as a nuclear counterstain. I've tried to use Eosin Yellow to stain the cytoplasm, but it doesn't seem to work. Any suggestions on staining protocol or alternative counterstains? I appreciate your inputs very much!
Just to perhaps help quickly: you could search in all RG-deposited/stored (Q&A) files for e.g. search words (this only proposals) via the RG-SEARCH-function (see menu bar find rectangle with "Search", mouse left click to the "downpointing arrow", from the pull down-menu choose "Questions" and insert your search word or search phrase):
' cytoplasmic counterstain ', ' Immunohistochemistry counterstain ', 'counterstain ICC', ' counterstain immunocytochemistry ' etc. etc. For your convenience I add here the URL which is created when searching for (e.g.): ' counterstain ICC IHC '
https://www.researchgate.net/search.Search.html?type=question&query=counterstain++++ICC++IHC++
You'll find plenty of Questions and Answers regarding your matter....and can go through until further replies/answers will be given to your specific request.... Best wishes and regards, WM
NB: it would be helpful if you tell the reader which ICC stain(ings, Staining 'system') you use/apply since the proposal of a suited cytoplasmic counterstain could depend on what you got prior to counterstaining (:-))
All depends on the color of the reaction product: Brown DAB I recommend use of a complementary counterstain that can be made invisible by filters (methyl green is wonderful in that it provides a good cytoplasmic stain but is hidden when photography with a green filter is used. For NiDAB stain (blue-black) the best is neutral Red in that it provides an excellent contrast and it can be made invisible by photography with a red filter. Fluorescent stains can be obtained that are cytoplasmic Nissl stains although I have not found them particularly useful in some of our sections.
Thank you, Wolfgang and Gloria for your answers! I'll look into the counterstains Methyl Green, Neutral Red and Cresyl Violet.
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