Objective In order to avoid potential injuries imposed to human body, it can be feasible to use the musculoskeletal models which can be reconstructed from the cadaver color cryosection (CCC) images, computerized tomography (CT) images, magnetic resonance (MR) images or other images to analyze the dynamic properties of muscles in vivo during human movement. Methods We reconstruct the lower limb musculoskeletal model and define the uniform joint coordinate system (JCS) on the model and the subject. The coordinate transformation of the muscle attachment points both on the model and the subject is described in detail. Results The length and the moment arm of the biceps femoris (short head) during knee flexion are calculated and analyzed. Conclusion This method plays an important role in improving the kinematics and dynamic simulation and the muscle force estimation.