We developed a protocol which determines age in spotted deer, Axis axis (Erxleben, 1777) through cementum layers in the tooth which is economical and requires lesser time (±19 hours). The permanent first incisor teeth from mandibles of known estimated ages (2, 4, 5, 8 and 9 years) were ground longitudinally on median plane on a set of sandpapers with hand to a thickness of around 57 μm, decalcified, stained with hematoxylin, differentiated in acid alcohol and mounted on a glass slide and count cementum layers under microscope at 100 or 200× magnification. Presence of 1, 3, 4, 7 and 8 cementum layers was observed for the tooth of ages of 2, 4, 5, 8 and 9 years, respectively. The final age in the year was calculated by adding one to the counted number of cementum layer. Our results were in compliance with the pre-estimated age of the mandibles. This suggested protocol saves time and money and is easy to perform with minimum requirements. The technique was validated using “blind” duplicate test (n = 2) and the first molar tooth of known estimated age (n = 1). Our suggested protocol has significant relevance for estimating the age of other animals and wildlife forensics.