Parameter calibration is the prerequisite and foundation to ensure the performance of robotic vision guidance systems, and the calibration accuracy directly determines the accuracy and reliability of robot vision guidance. In this chapter, we first demonstrate the camera intrinsic parameter calibration method based on the perspective projection imaging model. The extrinsic parameter calibration methods involved in binocular vision or multi-ocular vision systems are also introduced. By establishing the imaging model of the rotational-prism-based variable-boresight vision system, we propose two parameter calibration methods using reciprocating prism motion and flexible reference transfer, respectively. For hand-eye calibration between the robot coordinate system and the camera coordinate system, the basic equation for robot hand-eye relationship is derived. The classic two-step method and the auxiliary reference transfer strategy are presented to calculate the robot’s hand-eye matrix.