Spatially localized ballistic two-photon excitation in scattering media†

We describe spatially localized two-photon excitation in scattering media. Using femtosecond pulses at 770 nm from a Ti : Sapphire laser, we were able to excite fluorophores in capillary tubes under up to 1.5 mm of 0.5% intralipid. Displacement of the laser beam relative to the embedded samples indicates that highly localized excitation was possible with two-photon excitation, whereas one-photon excitation resulted in loss of spatial resolution due to excitation by the diffusely scattered photons. These results indicate that two-photon excitation in the scattering solution is due only to the ballistic photons, a result confirmed by frequency-domain time-resolved measurements. Selective excitation of adjacent embedded samples was found possible for two but not one-photon excitation. © 1998 John Wiley & Sons, Inc. Biospectroscopy 4: 303–310, 1998