Laser cladding on JK1002 NdYAG laser and the synchronous powder feeder are performed to fabricate micro-fluidic channels mold. Substrate material is HPM75 steel and clad powder is 316L stainless steel. Influences of the powder feed rate on clad layer geometrical dimensions and the combination of laser power and powder feed rate with stable state of powder feeding on clad quality are investigated. Results show that the range of 1.5～2.5 g/min of powder feed rate obtains micro clad layers which meet geometrical dimensions′ requirement of micro-fluidic channel mold and combination parameters of laser power of 400 W and powder feed rate of 2.0 g/min make micro clad layer with good clad quality. Scanning paths of micro-fluidic channel mold with specific shape are planned by CAD-Mastercam software. Using optimum parameters combination, multilayer laser cladding experiment is carried out and a sample of micro-fluidic channels mold is fabricated with dense structure and hardness of 580 HV. A little milling and polishing makes the sample meet the requirement of mold. Sizes of the final sample are 0.1 mm in height and 0.3 mm in width. The mold fabricating time is 60～75 min totally.