Loss of cell differentiation and increased cellular proliferative activity during malignant transformation leads to alteration of biochemical content of cells. This is reflected in the fluorescence profile of tissues. (1) To evaluate the efficacy of autofluorescence in clinical detection of oral cancer. (2) To correlate it with the rate of cell proliferation by analyzing argyrophilic nucleolar organizer regions (AgNORs). Autofluorescence status was studied by devising a visual enhancement system using ultraviolet light, followed by an incisional biopsy. Tissue was sent for fluorescence spectroscopy and analyzed by routine histopathology and AgNORs staining. The obtained results were statistically analyzed using Chi-square test (P < 0.05) and one-way analysis of variance. A statistically significant correlation was seen between autofluorescence (AF) and clinical diagnosis as well as autofluorescence and histopathological diagnosis. The importance of autofluorescence as a screening tool was further supported by a statistically significant correlation between autofluorescence status and cell proliferative rate. A preliminary effort was attempted at delving into this relatively unexplored arena of optical biopsy systems through this study. They can be used to monitor treatment and potential complications, surgical margins, detection of nodal metastasis, etc. They can provide a diagnosis noninvasively, in situ, and in real time.