Self-assembled multilayer thin films have been prepared on Au substrate by alternate surface derivatization with L-cysteine hydrochloride and cupric perchlorate. The layer-by-layer structure at each step of multilayer formation was investigated by X-ray photoelectron spectroscopy. The measurements indicate that there are two structure modes in the multilayers. One is that Cu2+ sandwiches between two amino acid groups. The other one is that Cu+ is bonded through disulfide and thiolate. This process is also confirmed by cyclic voltammetry of Cu ion at different self-assembled multilayers. Steps further on will lead to repeated multilayer films.