Tantalum pentoxide thin films are prepared by oblique angle electron beam evaporation. The influence of flux angle on the surface morphology and microstructure is investigated by scanning electron microscopy (SEM). The Ta2O5 thin films are anisotropic with highly orientated nanostructure of slanted columns. The porous microstructure of the as-deposited films results in the decrease of effective refractive index and packing density with increasing deposition angle. The anisotropic structure results in optical birefringence. The in-plane birefringence increases with the increase of deposition angle and reaches the maximum of 0.055 at the deposition angle of 70°. Anisotropic microstructure and critical packing density are the two key factors to influence the in-plane birefringence.