Spatial distribution of two-photon-excited fluorescence in scattering media.

Spatial distribution of two-photon-excited fluorescence (TPF) for dye beneath the surface of a highly scattering medium was investigated with picosecond laser pulses at 1064 nm. The active scattering media consisted of a suspension of polystyrene particles in a solution of Rhodamine 590 Tetrafluoroborate dye. With the increase of scattering strength of the medium, the location of the maximum TPF intensity was found to move closer to the surface away from the focal region; the intensity of TPF was not confined to the focal region as in the case of nonscattering medium but was more evenly distributed. The spatial resolution of nonlinear optical microscopy for probing a scattering medium is degraded. Taking into account the scattering of the medium qualitatively explains the observed TPF spatial distribution.