We combined mobile biplane X-ray imaging and magnetic resonance imaging to measure the regions of articular cartilage contact and cartilage thickness at the tibiofemoral and patellofemoral joints during six functional activities:​ standing, level walking, downhill walking, stair ascent, stair descent, and open-chain (non-weight-bearing) knee flexion. The contact centers traced similar paths on the medial and lateral femoral condyles, femoral trochlea, and patellar facet in all activities while their locations on the tibial plateau were more varied. The translations of the contact centers on the femur and patella were tightly coupled to the tibiofemoral flexion angle in all activities (r² > 0.95) whereas those on the tibia were only moderately related to the flexion angle (r² > 0.62). The regions of contacting cartilage were significantly thicker than the regions of non-contacting cartilage on the patella, femoral trochlea, and the medial and lateral tibial plateaus in all activities (p < 0.001). There were no significant differences in thickness between contacting and non-contacting cartilage on the medial and lateral femoral condyles in all activities, except open-chain knee flexion. Our results provide partial support for the proposition that cartilage thickness is adapted to joint load and do not exclude the possibility that other factors, such as joint congruence, also play a role in regulating the structure and organization of healthy cartilage. The data obtained in this study may serve as a guide when evaluating articular contact motion in osteoarthritic and reconstructed knees.