Frequency-domain lifetime measurements and sensing in highly scattering media

Abstract

For clinical chemistry, or for non-invasive sensing through skin, it is often necessary to obtain quantitative information in highly scattering media. We describe two simple methods for fluorescence lifetime measurements in highly scattering media, and in particular in an intralipid suspension. Lifetimes can be measured using an intensity decay law that accounts for the time delays and pulse-broadening effects of multiple light-scattering events in the intralipid. Alternatively, the phase and modulation measurements can be performed relative to a reference fluorophore with a known lifetime. These approaches provide reliable lifetime data for a pH-sensitive fluorophore contained within a microcuvette 4 mm under the surface of the intralipid suspension. Fluorescence lifetime-based sensing is now recognized as a valuable methodology in clinical and analytical chemistry, and the possibility of lifetime-based sensing in turbid media has been demonstrated.