The ultimate success of near-infrared optical tomography rests on the precise measurement of light propagation within tissues or random media, the accurate prediction of these measurements from a light propagation model, and an efficient three-dimensional solution of the inverse imaging problem. To date, optical tomography algorithms have focused on frequency-domain photon migration (FDPM) measurements of phase-delay and amplitude attenuation, which are reported relative to the incident light, even though phase-delay and amplitude of incident light are nearly impossible to measure directly. In this contribution, we examine referenced, fluorescence-enhanced frequency-domain photon migration measured at excitation and/or emission wavelengths and report on a measurement strategy to minimize measurement and calibration error for efficient coupling of data to a distorted Born iterative imaging algorithm. We examine three referencing approaches and develop associated inversion algorithms for (1) normalizing detected emission FDPM data to the predicted emission wave arising from a homogeneous medium, (2) referencing detected emission FDPM data to that detected at a reference point, and (3) referencing detected emission FDPM data to detected excitation FDPM data detected at a reference point. Our results show the latter approach to be practical while reducing the nonlinearity of the inverse problem. Finally, in light of our results, we demonstrate the method for eliminating the influence of source strength and instrument functions for effective fluorescence-enhanced optical tomography using FDPM.
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