To identify the optimal body surface puncture locations and the depths of nerve entry points (NEPs) in the deep posterior compartment muscles of the leg, 60 lower limbs of thirty adult cadavers were dissected in prone position. A curved line on the skin surface joining the lateral to the medial epicondyles of the femur was taken as a horizontal reference line (H). Another curved line joining the lateral epicondyle of the femur to the lateral malleolus was designated the longitudinal reference line (L). Following dissection, the NEPs were labeled with barium sulfate and then subjected to spiral computed tomography scanning. The projection point of the NEP on the posterior skin surface of the leg was designated P, and the projection in the opposite direction across the transverse plane was designated P'. The intersections of P on H and L were identified as P and P , and their positions and the depth of the NEP on PP' were measured using the Syngo system and expressed as percentages of H, L, and PP'. The P points of the tibial posterior, flexor hallucis longus and flexor digitorum longus muscles were located at 38.10, 46.20, and 55.21% of H, respectively. The P points were located at 25.35, 41.30, and 45.39% of L, respectively. The depths of the NEPs were 49.11, 54.64, and 55.95% of PP', respectively. The accurate location of these NEPs should improve the efficacy and efficiency of chemical neurolysis for treating spasticity of the deep posterior compartment muscles of the leg. Clin. Anat. 30:855-860, 2017. © 2017 Wiley Periodicals, Inc.