Blood smears

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cooper
cooper's picture
Blood smears

When I stain cancer tissue by ICC with my own monoclonal primary antibody, biotinylated secondary antibody, avidin/biotin complex, and DAB as substrate, I counterstain with hematoxylin.

I would next like to stain a blood smear using the same ICC method...is it possible to counterstain with both Wright-Giemsa and Myeloperoxidase??? If this is possible, at what point in my protocol should I perform these stains, and in what order? Currently, I stain with hematoxylin after the DAB step, followed by 1:10 glacial acetic acid, ammonia water, tap water, graded ethanol, and finally xylene. Would this work for the W-G and Myeloperoxidase stain? Thanks for your input!!!

Arvind Singh Pundir
Arvind Singh Pundir's picture
I have not tried this Giemsa

I have not tried this Giemsa counterstain but with my experience i would suggest that it is obvious that if you are doing IHC the counterstain step would be the last one and the counterstain should be in contrast to the chromogen used in IHC also the chemicals used in counterstaining should not react to your chromogen(DAB, NOVARED etc.) else you may loose the IHC signal, rest is that you try and standardise  the results
good luck