Digital image correlation (DIC) has been successfully applied for evaluating the mechanical behavior of biological tissues. A three-dimensional (3-D) DIC system has been developed and applied to examining the motion of bones in the human foot. To achieve accurate, real-time displacement measurements, an algorithm including matching between sequential images and image pairs has been developed. The system was used to monitor the movement of markers which were attached to a precisely motorized stage. The accuracy of the proposed technique for in-plane and out-of-plane measurements was found to be and 1.17%, respectively. Two biomedical applications were presented. In the experiment involving the foot arch, a human cadaver lower leg and foot specimen were subjected to vertical compressive loads up to 700 N at a rate of and the 3-D motions of bones in the foot were monitored in real time. In the experiment involving distal tibio fibular syndesmosis, a human cadaver lower leg and foot specimen were subjected to a monotonic rotational torque up to 5 Nm at a speed of 5 deg per min and the relative displacements of the tibia and fibula were monitored in real time. Results showed that the system could reach a frequency of up to 16 Hz with 6 points measured simultaneously. This technique sheds new lights on measuring 3-D motion of bones in biomechanical studies.