We performed a fluorescent microscopic examination of human and animal aortic grafts at different stages of decellularization. Treatment of aortic grafts with trypsin/EDTA solution for 48 h leads to their complete decellularization and preserved the connective tissue fiber backbone, which gains a netlike structure. The use of this protocol of decellularization leads to disappearance of subintimal calcium deposits in human aortic grafts. Differences in laser-induced fluorescence spectra of aortas before decellularization and at different stages of this process were revealed. Our findings suggest that the use of fluorescence induced by excimer lasers is promising for identification of the composition of biological tissues, analysis of their state, and the presence of changes.