The principle of optical tweezers array based on double-plate shearing interference and the particle separation chip by mircofluidic fabrication method is introduced. The function of particle trapping and separation of the optical tweezers array is verified by the experiments. According to the principle of multi-beam light interference, double-plate shearing interference can produce stripes with high brightness and sharp edge to insure the gradient force applied to the trapped particles. The high flexibility of the system is benefited from the easy adjustment of stripe period. The microfluidic chip made by glass is fabricated by chemical etching, with good optical property, mechanical strength and electricity insulation. And the glass microfluidic chip is suitable for the samples of cell and macromolecules, such as protein, because of weak adsorption of protein. Taking advantage of fewer sample consumption and higher efficiency, the combination of optical tweezers array with microfluidic chip could be one of important tools for micro-nano scale analytical technology in the near future.