The reliability of the estimate of joint kinematic variables and the relevant functional interpretation are affected by the uncertainty with which bony anatomical landmarks and underlying bony segment anatomical frames are determined. When a stereo-photogrammetric system is used for in vivo studies, minimising and compensating for this uncertainty is crucial. This paper deals with the propagation of the errors associated with the location of both internal and palpable femoral anatomical landmarks to the estimation of the orientation of the femoral anatomical frame and to the knee joint angles during movement. Given eight anatomical landmarks, and the precision with which they can be identified experimentally, 12 different rules were defined for the construction of the anatomical frame and submitted to comparative assessment. Results showed that using more than three landmarks allows for more repeatable anatomical frame orientation and knee joint kinematics estimation. Novel rules are proposed that use optimization algorithms. On the average, the femoral frame orientation dispersion had a standard deviation of 2, 2.5 and 1.5 degrees for the frontal, transverse, and sagittal plane, respectively. However, a proper choice of the relevant construction rule allowed for a reduction of these inaccuracies in selected planes to 1 degrees rms. The dispersion of the knee adduction-abduction and internal-external rotation angles could also be limited to 1 degrees rms irrespective of the flexion angle value.