The purpose of this study was to show the application of quantitative 4D CT for subtalar joint evaluation in healthy volunteers and cadavers. Fifteen healthy volunteers with no history of subtalar joint trauma and three cadavers were prospectively evaluated with dynamic CT. The subtalar joint was evaluated during a pronosupination cycle. All acquisitions (cadavers and healthy volunteers) were performed using intermittent sequential mode with a 320-MDCT scanner. Angles and distances between the talus and the calcaneus were measured semiautomatically. Measurement variation was described in healthy volunteers and in cadavers, the latter before and after resection of the cervical and interosseous ligaments. The mean effective dose was below 0.1 mSv. In cadavers, mean increases in joint amplitude over 19% and of 22% were seen after partial ligament sectioning and after full ligament sectioning, respectively. The interobserver variability of the measurement ratios was considered to be excellent for three of the measurements made (ICC > 0.87) and moderate for the fourth (ICC = 0.57). The normal range of joint motion in healthy volunteers is described, with joint amplitudes varying from 6.4% to 22.8%. Quantitative dynamic CT of the subtalar joint can provide a detailed analysis of joint motion, supporting its potential role in the evaluation of subtalar instability.