Fluorescence-lifetime determination in tissues or other scattering media from measurement of excitation and emission kinetics.

ABSTRACT

Measurements of nanosecond and subnanosecond fluorescence lifetimes are restricted to dilute, nonscattering systems since excitation and emission photon times of flight significantly affect measured fluorescent decay kinetics. We provide the theoretical rationale for frequency-domain measurements of phase-shift and amplitude demodulation made at excitation and emission wavelengths for direct determination of lifetimes in tissues and other scattering media. We confirm our analytical expressions using standard laser dyes such as 3,3'-diethylthiatricarbocyanine iodide, IR- 125, and IR- 140 in polystyrene suspensions with similar scattering properties as tissues. Our results have significant implication for lifetime-based spectroscopy in tissues and other scattering media.