We report on the use of time-resolved and polarised evanescent wave-induced fluorescence anisotropy measurements to probe molecular photophysics, motion and energy migration of fluorescent species in close proximity to a silica/film interface. In particular we show that the fluorescence decay and anisotropy of common fluorophores varies as a function of the plane of the fluorophore with respect to the interface, the distance from the interface, and as a function of position (using polarised EW imaging). We have applied time-resolved and polarised EW-induced fluorescence microspectroscopic measurements to dyes, thin polymer nanoparticle films and cells on silica surfaces, probing the variation in the photophysical dynamics within the films.
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