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Effects of scattering in layered biological tissue on imaging spectroscopy data

Spectral imaging permits two-dimensional mapping of the reflectance properties of biological systems. However, imaging in turbid media involves pixel sizes that are comparable to or smaller than the mean photon path length. This implies that the spectrum measured at a given pixel in the image plane will be determined by manifold photon trajectories through an extended volume in the object, so there is not a uniquely defined path length. In addition, this implies nonlinear spectral mixing for systems with multiple layers and chromophores. Using Monte Carlo model, we have studied photon path distributions in the case of layered turbid systems and their effects on spectral imaging. In particular, we emphasize the effect of hemoglobin on imaging reflectance-mode hyperspectral data.