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A complete mathematical framework for preclinical optical imaging (OI) support comprising bioluminescence imaging (BLI), fluorescence surface imaging (FSI) and fluorescence optical tomography (FOT) is presented in which optical data is acquired by means of a microlens array (MLA) based light detector (MLA-D). The MLA-D has been developed to enable unique OI, especially in synchromodal operation with secondary imaging modalities (SIM) such as positron emission tomography (PET) or magnetic resonance imaging (MRI). An MLA-D consists of a (large-area) photon sensor array, a matched MLA for field-of-view definition, and a septum mask of specific geometry made of anodized aluminum that is positioned between the sensor and the MLA to suppresses light cross-talk and to shield the sensor's radiofrequency interference signal (essential when used inside an MRI system). The software framework, while freely parameterizable for any MLA-D, is tailored towards an OI prototype system for preclinical SIM application comprising a multitude of cylindrically assembled, gantry-mounted, simultaneously operating MLA-D's. Besides the MLA-D specificity, the framework incorporates excitation and illumination light-source declarations of large-field and point geometry to facilitate multispectral FSI and FOT as well as three-dimensional object recognition. When used in synchromodal operation, reconstructed tomographic SIM volume data can be used for co-modal image fusion and also as a prior for estimating the imaged object's 3D surface by means of gradient vector flow. Superimposed planar (without object prior) or surface-aligned inverse mapping can be performed to estimate and to fuse the emission light map with the boundary of the imaged object. Triangulation and subsequent optical reconstruction (FOT) or constrained flow estimation (BLI), both including the possibility of SIM priors, can be performed to estimate the internal three-dimensional emission light distribution. The framework is susceptible to a number of variables controlling convergence and computational speed. Utilization and performance is illustrated on experimentally acquired data employing the OI prototype system in stand-alone operation, and when integrated into an unmodified preclinical PET system performing synchromodal BLI-PET in vivo imaging.
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