In certain embodiments, the invention relates to systems and methods for altering an image to compensate for variation in one or more physical and/or supervenient properties (e.g., optical absorption and/or scattering) in heterogeneous, diffuse tissue, thereby attenuating the effects of tissue waveguiding. The methods enable the proper identification of emission image regions that evidence waveguiding of electromagnetic radiation, and enables compensation of emission images for such waveguiding. The methods preserve the depth localization accuracy of the FMT approach and improve optical reconstruction in the targeted areas while eliminating spurious components of fluorescence from the acquired data set. Calibration methods for probe concentration mapping are also presented.
