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Vibrational spectroscopic imaging has become an essential tool for tissue analyses in life science and represents a modern analytical technique enabling the detection and characterization of molecular components of biological samples. It is based on the absorption of IR radiation by vibrational transitions in covalent bonds and enables global analysis of samples, with resolution close to the cellular level. Advantage of vibrational spectroscopic imaging is the acquisition of local molecular expression profiles, while maintaining the topographic integrity of the tissue by avoiding time-consuming extraction, purification and/or separation steps, respectively. With this nondestructive analytical method it is possible to get both qualitative and quantitative information of heterogeneous samples and unique chemimorphological information about the tissue status, which represents an important benefit for future analytical interpretation of pathological changes of a tissue.