Monolayer Mo films with the same thickness are deposited on the Si substrates by direct current pulse magnetron sputtering method. The influence of different deposition rates on the microstructure, morphology and optical properties of Mo films is investigated with stylus surface profiling system, X-ray diffractometer, field emission scanning electron microscopy, atomic force microscopy and ultraviolet-visible spectrophotometer, respectively. The Mo film thickness fitted by grazing incidence X-ray reflective spectra agrees well with the designed thickness, which indicates that the current process technology of Mo film is mature and stable. The morphology evolution of Mo films is affected by the deposition rate via altering the growth model and nucleation rate of Mo films. As the increase of deposition rate, the density, diffractive intensity of Mo (110) and average grain size of films increase, whereas the surface roughness decreases first and then increases.