Background: The syndesmosis ligament complex stabilizes the distal tibiofibular joint while allowing for small amounts of physiologic motion. When injured, malreduction of the syndesmosis is the most important factor that contributes to inferior functional outcomes. Syndesmotic reduction is a dynamic measure, which is not adequately captured by conventional computed tomography (CT). Four-dimensional CT (4DCT) can image joints as they move through range of motion (ROM). The aim of this study was to employ 4DCT to determine in vivo syndesmotic motion with ankle ROM in uninjured ankles. Methods: Uninjured ankles were analyzed in patients who had contralateral syndesmotic injuries, as well as a cohort of healthy volunteers with bilateral uninjured ankles. Bilateral ankle 4DCT scans were performed as participants moved their ankles between maximal dorsiflexion and plantarflexion. Multiple measures of syndesmotic width, as well as sagittal translation and fibular rotation, were automatically extracted from 4DCT using a custom program to determine the change in syndesmotic position with ankle ROM. Results: Fifty-eight ankles were analyzed. Measures of syndesmotic width decreased by 0.7 to 1.1 mm as the ankle moved from dorsiflexion to plantarflexion ( P < .001 for each measure). The fibula externally rotated by 1.2 degrees with ankle ROM ( P < .001), but there was no significant motion in the sagittal plane ( P = .43). No participants with bilateral uninjured ankles had a side-to-side difference in syndesmotic width of 2 mm or greater. Conclusion: 4DCT allows accurate, in vivo syndesmotic measurements, which change with ankle ROM, confirming prior work that was limited to biomechanical studies. Side-to-side syndesmotic measurements are consistent within subjects, validating the method of templating syndesmotic reduction off the contralateral ankle, in a consistent ankle position, to achieve anatomic reduction of syndesmotic injury. Level of Evidence: Level II.