In order to improve the HV hardness and microstructure on the surface of TC4 titanium alloy, the high frequency scan of electron beam (EB) was introduced to treat TC4 titanium alloy. For excellent parameters, the EB energy, focus current and scan times, which will influence the HV hardness and microstructures, were studied. After high frequency scan of electron beam treatment, the HV hardness, modified layer depth and microstructures of TC4 titanium alloy were analyzed. Results show that the HV hardness on the TC4 surface treated by the high frequency scan of electron beam can be increased by almost 40 MPa compared with that of the original material. With improved energy of EB, the depth layer influenced is increased. The focus current adjusted in a narrow range can acquire comparatively better HV hardness on the surface. When the scan times are increased, the HV hardness decreases, and the grains grow up.