Antibodies optimal fluorophore

A series of antibody dilution tests prior to the combined ISH/ IHC can give a good hint of optimal antibody dilution. Best fluorescence images are obtained when the intensity of the individual fluorophores are balanced. 

Before using a fluorophore-labeled antibody from flow cytometry for immunocytochemistry, check the spectra of fluorescent labels to rule out the possibility of bleed-through. Remember that although flow cytometry and immunocytochemistry use fluorescent dyes, the characteristics of these dyes are optimized for the technique and might not translate well when moved from flow cytometry to immunocytochemistry. 

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