To determine the mechanisms and extents of popliteus impingements before and after TKA and to investigate the influence of implant sizing. The hypotheses were that (i) popliteus impingements after TKA may occur at both the tibia and the femur and (ii) even with an apparently well-sized prosthesis, popliteal tracking during knee flexion is modified compared to the preoperative situation. The location of the popliteus in three cadaver knees was measured using computed tomography (CT), before and after implantation of plastic TKA replicas, by injecting the tendon with radiopaque liquid. The pre- and post-operative positions of the popliteus were compared from full extension to deep flexion using normosized, oversized and undersized implants (one size increments). At the tibia, TKA caused the popliteus to translate posteriorly, mostly in full extension: 4.1mm for normosized implants, and 15.8mm with oversized implants, but no translations were observed when using undersized implants. At the femur, TKA caused the popliteus to translate laterally at deeper flexion angles, peaking between 80º-120º: 2.0 mm for normosized implants and 2.6 mm with oversized implants. Three-dimensional analysis revealed prosthetic overhang at the postero-superior corner of normosized and oversized femoral components (respectively, up to 2.9 mm and 6.6 mm). A well-sized tibial component modifies popliteal tracking, while an undersized tibial component maintains more physiologic patterns.