BackgroundTo optimize the biomechanical outcomes in lateral ankle ligament reconstruction, avoid stiffness or residual laxity, aiming for an isometric reconstruction of the anterior lateral talofibular ligament (ATFL) and the calcaneofibular ligament (CFL) is mandatory. However, the localization of the optimal ligament insertion remains challenging to assess intraoperatively. MethodThree-dimensional (3D) surface models from 10 healthy ankles were generated. 30 insertion points of the CFL were defined on the lateral side of the calcaneus each 10% of its total length in the dorsal-to-ventral and proximal-to-distal plane. 6 insertion points were defined at the ventral ridge of fibula from the malleolar tip and 5 insertions were defined along the lateral talar process. The ligament length variation of ATFL and CFL was assessed after a simulation of the flexion/extension around a simulated tibiotalar axis and inversion/eversion around a simulated subtalar axis in 36 different positions. ResultsThe isometric point of CFL on the calcaneus is located at about 60% along the dorsal-to-ventral and between 60% and 70% along the proximal-to-distal plane. From maximal extension to flexion, these points present respectively a length variation of − 0.8 to − 1.1 mm (p = 0.46) and − 1.1 to − 0.8 mm (p = 0.56). A fibular insertion at 5 mm proximal to the malleolar tip present a length variation ranging from − 0.1–1 mm (p < 0.001) for ATFL and from − 0.7–0.5 mm (p < 0.001) for CFL. A talar insertion point of the ATFL located 5 mm proximal to the subtalar joint present the lowest variation, ranging from − 1.1–0.7 mm (p < 0.001), however an insertion at 20- or 25-mm present isometry (+0.1 to +0.9 mm p = 0.1, and +0.4 to +0.4 mm p = 1 respectively) if the fibular insertion is located at 5 mm proximal to the malleolar tip. ConclusionThis study provides anatomical references which are reproducible in daily practice. These insertion points allow to achieve a stable reconstruction while maintaining a tension-free mobilization of the ankle.