Bioluminescence in vitro studies are usually performed with dedicated microscopes. In this work, we proposed a novel image recovery algorithm and a multimodal system prototype to perform bioluminescence microscopy. A super resolution regularization algorithm (SRRA) was developed and applied in a feasibility study using Geant4 Monte Carlo (MC) simulation. The method was also tested using in vitro cells acquisition performed with the prototype. The results obtained with MC simulations showed an improvement in the spatial localization resolution from 90 μm to 10 μm. The processing of the in vitro mesothelioma cells image shows shrink of the localization full width half maximum (FWHM) from 110 μm to 13 μm. © 2015 IEEE.
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