Background: Teeth can survive and remain virtually unaffected long after other soft tissue and skeletal tissues have been destroyed. The present study was conducted to assess pulp tissue in gender determination. Materials & Methods: 30 periodontally or endodontically extracted premolar teeth of both genders were further categorized into 5 groups of 6 each (3 from males and 3 from females) based on the extent of caries progression. Group I includes freshly extracted tooth specimens with no caries, group II consisted of tooth specimens with caries in enamel, group III comprised of tooth specimens with caries less than half way of dentin, group IV consisted of freshly extracted teeth with caries more than half way of dentin, and group V includes freshly extracted tooth specimen with caries involving pulp. The pulp cells are stained with quinacrine hydrochloride and observed with fluorescent microscope for fluorescent body. Gender was determined by identification of Y chromosome fluorescence in dental pulp. Results: Efficiency of diagnostic utility of fluorescent body in identifying gender in group I was 100%, in group II was 100%, in group III was 80%, in group IV was 50% and in group V was 45%. The percentage of fluorescent bodies in males and females in group I was 70.4 and in group II was 4.7, in group II was 64.2 and 5.3, in group III was 40.5 and 18.5, in group IV was 22.6 and 16.4 and in group V was 10.2 and 12.9 respectively. Conclusion: When determining the sex of a tooth with healthy pulp tissue, caries with enamel or dentin up to half of its length, fluorescence staining of the Y chromosome is a dependable method. Teeth with pulp-related caries cannot be used to determine a person's sex.