A simple method for detecting up to five immunofluorescent parameters together with DNA staining for cell cycle or viability on a benchtop flow cytometer.

In this manuscript, we describe modifications to a commercial three-laser benchtop flow cytometer, as well as relevant biological methods, that allow analysis of up to five immunofluorescent parameters together with an ultraviolet (UV)-excitable DNA stain. This method allows expanded capacity for multiparameter immunophenotyping of complex mixed cell populations, together with accurate measurements of DNA content (cell cycle) or cell viability, on a stable, end-user operated platform.