Bicruciate-retaining total knee arthroplasty (BCR-TKA) recreates normal knee movement by preserving the anterior cruciate and posterior cruciate ligaments. However, in vivo kinematics of BCR-TKA with the anatomical articular surface remains unknown. The objective of this study was to evaluate in vivo kinematics of BCR-TKA with the anatomical articular surface during high-flexion activities. For this purpose, 17 knees after BCR-TKA with an anatomical articular surface were examined. Under fluoroscopy, each patient performed squatting and cross-legged sitting motions. To estimate the spatial position and orientation of the knee, a two-dimensional or three-dimensional registration technique was used. Rotation, varus-valgus angle, and anteroposterior translation of medial and lateral contact points of the femoral component relative to the tibial component were evaluated in each flexion angle. The results showed that from 80 to 110° of flexion, the femoral external rotation during squatting was significantly larger than that during cross-legged sitting. At maximum flexion, the knees during sitting indicated significantly more varus alignment than during squatting. During squatting, a medial pivot pattern was observed from minimum flexion to 10° flexion, with no significant movement beyond 10° of flexion. Conversely, during cross-legged sitting, no significant movement was detected from minimum flexion to 60° of flexion, with a medial pivot beyond 60° of flexion. Therefore, the knees showed relatively normal kinematics after BCR-TKA with an anatomical articular surface; however, it varied during high-flexion activities depending on the activity.