Despite decades of research since its first description, subtalar joint instability remains a diagnostic enigma within the concept of hindfoot instability. This could be attributed to current imaging techniques, which are impeded by two-dimensional measurements. Therefore, we used weightbearing computed tomography imaging to quantify three-dimensional displacement associated with subtalar joint instability. Three-dimensional models were generated in seven paired cadaver specimens to compute talocalcaneal displacement after different patterns of axial load (85 kg) combined with torque in internal and external rotation (10 Nm). Sequential imaging was repeated in the subtalar joint containing intact ligaments to determine reference displacement. Afterward, the interosseus talocalcaneal ligament (ITCL) or calcaneofibular ligament (CFL) was sectioned, then the ITCL with CFL and after the ITCL, CFL with the deltoid ligament (DL). The highest translation could be detected in the dorsal direction and the highest rotation occurred in the internal direction when external torque was applied to the foot without load. These displacements differed significantly from the condition containing intact ligaments, with a mean difference of 1.6 mm (95% CI, 1.3 to 1.9) for dorsal translation and a mean of 12.4° (95% CI, 10.1 to 14.8) for internal rotation. Clinical relevance: Our study provides a novel and noninvasive analysis to quantify subtalar joint instability based on three-dimensional WBCT imaging. This approach overcomes former studies using trans-osseous fixation to determine three-dimensional subtalar joint displacement and implements an imaging device and software modalities that are readily available. Based on our findings, we recommend applying torque in external rotation to the foot to optimize the detection of subtalar joint instability.