Purpose: Exact knowledge of the location and dimension of the pulp chamber help to maintain the pulp healthy during operative procedure and also reduces the risk of perforation of pulp chamber during root canal treatment. This in-vivo study was carried out to measure critical morphology of pulp chamber of mandibular molar using intra-oral periapical radiograph. Materials and Methods: Mandibular molar teeth of 56 patients were evaluated. The mandibular molar teeth whose pulp chamber was not violated by caries, restoration, fracture crown and those having closed apex were included in the study. The intraoral periapical radiographs were taken with paralleling angle technique using radio-opaque grid with 1 mm space. This grid was placed directly on the film. Results: In 94% of the mandibular first molars specimens the pulp chamber ceiling was at the level of the cemento-enamel junction. The measurements showing the lowest percentage variance were buccal cusp to furcation (approximately 11%) and buccal cusp to pulp chamber ceiling (approximately 15%). The distance from the cusp tip to pulp chamber ceiling height was approximately 6.0 mm, the distance from the pulpal floor to the furcation was approximately 3.0 mm, and the average height of a pulp chamber was 1.5 to 2.0 mm. Conclusion: The exact knowledge of distances of pulp chamber from various anatomical landmarks helps in proper assessment of root canals and ultimately avoids the failure of root canal treatment.