Fluorescence Polarization as a Functional Parameter in Monitoring Living Cells: Theory and Practice

The use of fluorescence polarization as a functional parameter in monitoring cellular activation calls for the reliable and accurate measurement of the fluorescence intensity and polarization (FI and FP) of microscopic objects. The relevant experimental parameters that enter such measurements are thoroughly discussed. The possibility of executing FP measurements properly by flow-through systems is compared with that of static cytometry. Remarks on the effects of high-power excitation on markers and cells conclude the paper.