In this paper, we developed a microfluidic chip based method for fabrication of bio-actuators driven by artificial muscles made from molecular motors. Different from conventional complex methods, we integrates the fabrication and actuation of bio-actuators into a single microfluidic chip using UV-induced projection printing with a maskless optical system. A micro-gripper was fabricated and actuated in a microfluidic chip as an example of application. This low-cost and easy-operated method is able to make different kinds of bio-actuators for microfluidic devices.