Experimental Conditions and Mathematical Analysis of Kinetic Measurements Using Flow Cytometry – The FacsKin Method

Flow cytometry is in use for the assessment of different cell subsets' prevalence for decades. However, the development of specific dyes sensitive for the quickly changing intracellular analytes provided an opportunity for the real-time monitoring of intracellular processes with flow cytometry. In these kinetic measurements additionally to cell prevalence values the time as a novel variable is introduced. This enables researchers to gather data on cellular functionality from a new perspective, as the recording of a kinetic parameter with flow cytometry provides information on intracellular processes in several cell subtypes in a simultaneous manner, where physiological cell-cell interactions (such as cytokines) are still present. This approach may be a useful tool in many different areas of immune research. However, the mathematical formulae to characterize the several millions of data recorded during one measurement are still missing.