Abstract: This invention provides a system and method for calibration of a 3D vision system using a multi-layer 3D calibration target that removes the requirement of accurate pre-calibration of the target. The system and method acquires images of the multi-layer 3D calibration target at different spatial locations and at different times, and computes the orientation difference of the 3D calibration target between the two acquisitions. The technique can be used to perform vision-based single-plane orientation repeatability inspection and monitoring. By applying this technique to an assembly working plane, vision-based assembly working plane orientation repeatability, inspection and monitoring can occur. Combined with a moving robot end effector, this technique provides vision-based robot end-effector orientation repeatability inspection and monitoring. Vision-guided adjustment of two planes to achieve parallelism can be achieved.