Abstract: To investigate the feasibility of Microsoft Kinect camera for guiding percutaneous liver needle intervention, a puncture navigation system based on the second generation Kinect was developed. The two surfaces from preoperative computed tomograph (CT) and intraoperative Kinect RGB-depth images were extracted and matched for physical-to-image registration. Experiments were conducted on a general abdominal phantom and six beagles in vivo. The target registration error, user error, and target positioning error were used to evaluate navigation accuracy. The results show that target registration error, user error, and target positioning error of the phantom are (4.26±1.94) mm, (2.92±1.67) mm and (5.23±2.29) mm, respectively. In addition, two generations of Kinect were both tested, and the results imply that the second generation Kinect-based navigation is superior to the first-generation. For the animal experiment, the target positioning error was evaluated as (6.40±2.72) mm, and its lateral and longitudinal component were (4.30±2.51) mm and (3.80±3.11) mm, respectively. This study demonstrates that the Kinect camera has a potential of clinical application in percutaneous liver puncture navigation.