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Deposit Ti film on polished Mo, quartz and single crystal silicon subserate by using electron beam evaporation. Analized the surface morphology and microstructures of substrates and the thin film by AFM, XRD and SEM.The results shown that : The Ti film’s surface morphology and microstructure were greatly influenced by the substrate materials. The surface of the Ti film on the polished MO substrates had some fluctuations. The Ti film nucleated on the substrate firstly, and then vertically grown upwards in the form of columnar grain. The surface of Ti film on the polished quartz substrate was smooth, and the particles and the interfaces were clearly distinguished by the layer of equiaxed grains on the interface. The surface of Ti film on the single crystal silicon substrate was the roughest one. However, the surface of the crystal silicon was the smoothest one. There were peaks of TiSi2 controlled by Ti/Si interface silicide growth dynamics.