The properties and structure of the crystalline lens change as time after death passes. Some experiments have suggested that these might be used to estimate the postmortem interval (PMI). In this study, the organization and texture of the rabbit lens were objectively evaluated as a function of the PMI using two-photon excitation fluorescence (TPEF) imaging microscopy. Between 24 h and 72 h, the lens presented a highly organized structure, although the fiber delineation was progressively vanishing. At 96 h, this turned into a homogeneous pattern where fibers were hardly observed. This behaviour was similar for parameters providing information on tissue texture. On the other hand, the fiber density of the lens is linearly reduced with the PMI. On average, density at 24 h was approximately two-fold when compared to 96 h after death. The present results show that TPEF microscopy combined with different quantitative tools can be used to objectively monitor temporal changes in the lens fiber organization after death. This might help to estimate the PMI, which is one of the most complex problems in forensic science.
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