The influence of the substrate temperature on the topography and microstructure of the diamond-like carbon (DLC) film in the pulsed laser deposition (PLD) is studied. When the diamond-like carbon films were deposited,the substrate temperature kept at 30,200,400 and 600 ℃ respectively. The microstructure and composition of the diamond-like carbon films deposited at different substrate temperatures were detected by the visible Raman spectroscopy and the X-ray photoelectron spectroscopy (XPS) respectively. The topography of the diamond-like carbon films was detected by the atomic force microscopy (AFM). The experimental result demonstrates that the substrate temperature influences microstructure,composition,ratio of sp3/sp2 and topography of the diamond-like carbon films in the pulsed laser deposition remarkably. When the substrate temperature increases,the ratio of sp3/sp2 in the film would decrease,the size and number of graphitic crystallites in the film would increase,the roughness of the film would reduce and the number of granules on the film would decrease. The reason to cause the experimental phenomenon is that transfer ability of the carbon ions and granules on the substrate would increase when the substrate temperature increases.