Micro-assembling Teflon/Si3N4 multilayer film was developed by ion beam alternating sputtering Teflon and Si3N4 ceramic targets. The structural, mechanical and microtribological properties were studied by PHI-5300, FTIR, XRD and atomic force and friction force microscope (AFM/FFM). The results show that the multilayer consists of Si3N4 component and crystalline Teflon. The hardness of the multilayer is less than that of Si3N4; but the toughness of Teflon/Si3N4 is greatly improved. The friction coefficient of Teflon/Si3N4 multilayer is lower than that of Si3N4 film, and the wear resistance of Teflon/Si3N4 multilayer is much greater than that of Teflon film. The friction force of Teflon/ Si3N4 film is linear with the load in nanoscale. The worn track will be formed in Teflon and Teflon/Si3N4 film when the load is greater than 70 nN.