A series of Hafnium dioxide (HfO2) thin films with nominal thickness of 5–350 nm were reactively deposited on silicon(100)substrates at 200 °C using electron-beam evaporation. Based on the measurement and characterization techniques of spectroscopic ellipsometry, X ray diffraction, scanning electron microscopy and atomic force microscopy, the effect of growth thickness on the refractive index and crystallization of HfO2 film grown under the same conditions was studied. The results indicate that the change of refractive index of HfO2 film is closely related to the change of microstructure i.e., grain size and crystallization which change obviously with the increasing growth thickness. The average refractive index at 633 nm decreases about from 1.97 to 1.84 with the increase of the average size about from 0.5 nm to 7~8 nm and the crystallinity from zero to 74% when the thickness of HfO2 film increases from 5 nm to 350 nm. Meanwhile, a critical growth thickness of HfO2 film, somewhere on order of 130 nm, is confirmed at which the film transforms from an amorphous to a polycrystalline monoclinic structure. The sharp change of microstructure near the critical thickness value of HfO2 film leads to the abrupt change of optical properties of HfO2 film, such as the turning behaviors of the refractive index curve. The correlation between phase transformation, size and orientation of crystallite, packing density and hence the refractive index is established. The possible interpretations are proposed to well understand the underlying mechanism of the evolution of optical properties in HfO2 film-forming process.