Abstract: Compared with the traditional minimally invasive surgery, the preoperative preparation of the robot-assisted minimally invasive surgery is more critical and complex. For the purposes of improving the operation efficiency and making full use of the characteristics of the robot, the kinematic performance of the manipulator as well as the collaborative capability between instruments are taken as the optimization objectives, and a new dexterity index named IICV for robot-assisted minimally invasive surgery and the internal collaboration space index named IICS are proposed. A set of multi-objective preoperative optimization methods based on NSGA-II are designed to optimize the incision placement and initialize the pose of the manipulator as a unified whole. Finally, a comparison between the preoperative planning schemes generated by the optimization algorithm and recommended by an experienced surgeon is performed. The experimental result shows that the preoperative planning scheme obtained by the optimization algorithm is effective, and it can provide a relatively superior operating environment for robot-assisted minimally invasive surgery.