I used IHC method with biotinylated primary antibody for my protein of interest which is ubiquitously present in the tissue. I am developing a protocol. I used DAB for 2 mins throughout my experiments. I tried different fixative (different hours), different antigen retrieval method and even different concentrations of primary antibody and ABC vectastain as well.

I am now to analyse these, I took the images but how to analyse these? I need some opinion on that :)

Thank you in advance