The subtalar joint is important in frontal plane movement and posture of the hindfoot. Abnormal subtalar joint moments caused by muscle forces and the ground reaction force acting on the foot are thought to play a role in various foot deformities. Calculating joint moments typically requires knowledge of the location of the joint axis; however, location of the subtalar axis from measured movement is difficult because the talus cannot be tracked using skin-mounted markers. The accuracy of a novel technique for locating the subtalar axis was assessed in vivo using magnetic resonance imaging. The method was also tested with skin-mounted markers and video motion analysis. The technique involves applying forces to the foot that cause pure subtalar joint motion (with negligible talocrural joint motion), and then using helical axis decomposition of the resulting tibiocalcaneal motion. The resulting subtalar axis estimates differed by 6 degrees on average from the true best-fit subtalar axes in the MRI tests. Motion was found to have been applied primarily about the subtalar joint with an average of only 3 degrees of talocrural joint motion. The proposed method provides a potential means for obtaining subject-specific subtalar axis estimates which can then be used in inverse dynamic analyses and subject-specific musculoskeletal models.