The multilayered film on a glass substrate, assembled from cationic poly(ethyleneimine) (PEI), anionic nano-sized polystyrene (PS) latex particle and cationic PEI or poly(allylamine hydrochloride) (PAH) layer was prepared by using the layer-by-layer (LbL) technique. Hydrophobization was carried out by immersing the film in Nafion solution. The hydrophobicity with high contact angle value was achieved by adjusting deposition ionic strength and ratio of water/ethanol of Nafion solution. The existence of PS particles in the LbL film significantly enhanced hydrophobicity of the film surface.