The liver, due to its central role in the regulation of metabolism and multiplicity of its other functions, is exposed to a high number of pathogenic factors. Non-Alcoholic Fatty Liver Disease (NAFLD), manifested mainly by changes in liver's lipid profile, represent one of the most frequently occurring liver pathology, that is often underestimated at an early stage of its development. The pathogenesis as well as the treatment of NAFLD has not been fully established yet. The difficulty associated with this arises from the need to detect very subtle changes. With regard to fulfilling these expectations, from all available research methods vibrational spectroscopic imaging techniques attract special attention, as they possess a great potential to detect even minor variation in chemical composition of the sample. The specificity of infrared imaging and Raman mapping along with their complementarity provides the ability to obtain comprehensive information on the molecular level. Here we present an approach for the study of liver tissue based on the simultaneous application of vibrational spectroscopic imaging and multirivate data analysis combined with histochemical staining.