CO2 laser rapidly patterning microfluidic chips on polymer substrate with direct writing ablation had attracted many experts from domestic and foreign in resent years. This method had features of flexible, rapid, easily controllable and high efficient, and was regarded as the most potentialized fabrication method to improve the development of microfluidic systems. This paper systematically reviewed polymer substrate materials, the mechanism of laser and material, fabrication models and factors influenced the fabrication process, as well as the influence on microfluidic chips fabrication quality by laser process parameters. Moreover, the existent problems and possible resolutions based on the mechanisms of interactions between laser and materials was analyzed.