Global analysis of dynamic fluorescence anisotropy by a polarized phasor approach.

Recently, the graphical analysis of the fluorescence lifetime imaging using the phasor approach has been highlight, and a series of the reports have made it on the way for the applications by the nonprofessionals. In this paper, we put forward a similar theory validated by the experiments for the dynamic fluorescence anisotropy imaging. By subtracting the perpendicular component from the parallel one in the frequency-domain polarization measurement, we deduce a new analytical expression about the fluorescence joint time, and find that as much as the fluorophore is a single exponential decay and r∞ is equal to zero, △I(t) is a single exponential decay with the time constant X as well, and the center of its histograms is located on the semicircle in the polarized phasor plot. In the end, we conclude that the fluorescence joint time is the best parameter to weigh the fluorescence dynamics for the macromolecules.