The knowledge of normal patellar tracking is essential for understanding the knee joint function and for diagnosis of patellar instabilities. This paper investigated the patellar tracking and patellofemoral joint contact locations during a stair ascending activity using a validated dual-fluoroscopic imaging system. The results showed that the patellar flexion angle decreased from 41.9° to 7.5° with knee extension during stair ascending. During first 80% of the activity, the patella shifted medially about 3.9 mm and then slightly shifted laterally during the last 20% of the ascending activity. Anterior translation of 13 mm of the patella was measured at the early 80% of the activity and the patella slightly moved posteriorly by about 2mm at the last 20% of the activity. The path of cartilage contact points was slightly lateral on the cartilage surfaces of patella and femur. On the patellar cartilage surface, the cartilage contact locations were about 2mm laterally from heel strike to 60% of the stair ascending activity and moved laterally and reached 5.3mm at full extension. However, the cartilage contact locations were relatively constant on the femoral cartilage surface (∼5mm lateral). The patellar tracking pattern was consistent with the patellofemoral cartilage contact location pattern. These data could provide baseline knowledge for understanding of normal physiology of the patellofemoral joint and can be used as a reference for clinical evaluation of patellofemoral disorders.