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Fourier-Transform Infrared Spectroscopy (FTIR) employs a unique approach to optical diagnosis of tissue pathology based on the characteristic molecular vibrational spectra of the tissue. The biomolecular changes in the cellular and sub-cellular levels developing in abnormal tissue, including a majority of cancer forms, manifest themselves in different optical signatures, which can be detected in infrared microspectroscopy. This report has two parts. In the first part, we report studies on normal, premalignant (polyp) and malignant human colonic tissues from three patients with different stages of malignancy. Our method is based on microscopic infrared study (FTIR-microscopy) of thin tissue specimens and a direct comparison with traditional histopathological analysis, which serves as a “gold” reference. The limited data available showed normal colonic tissue has a stronger absorption than polypoid tumor and cancerous types over a wide region in a total of 100 measurements. Detailed analysis showed that there is a significant decrease in total carbohydrate, phosphate and possibly creatine contents for polyp and cancerous tissue types in comparison to the controls. The same trend is maintained in seven other patients studied. The second part consists of an analysis showing the influence of various independent factors such as age, sex and grade of malignancy. Our preliminary results suggest that among the above three factors, age and grade of malignancy have significant effect on the metabolites level, but sex has only minor effect on the measured spectra. Initial results on Linear Discriminant Analysis (LDA) showed good classification between normal and malignant cells of human colonic tissues.