Abstract: In consideration of the shortcomings of traditional laser micro-manipulators, a laser micro-manipulator with the reflection structure was designed to improve the quality and reliability of spots. By taking a He-Ne laser as its indication light beam and a CO2 laser as its working light beam, the repeatability and sizes of the two spots were optimized. Furthermore, using the multi-configuration of a ZEMAX, the adjustable performance of working distance was realized from 200 mm to 400 mm. By carring out the corresponding experiment, then the optical system chart, diffraction encircled energy diagram and the spot diagram were given. The experimental results show that the repeatability of spots and the uniformity, stability and reliability of beams are improved a lot. With the characteristics of its clear operational visual field and treated safety, the laser micromanipulator can be combined with the operation microscopy to meet the long-term use need of the treatment.