3-Mercaptopropionic acid-stabilized CdTe nanoparticles were prepared and assembled layer-by-layer with poly(diallyidimethylammonium chloride) (PDAC) to form a polymer-supported ultrathin film by virtue of the Coulombic interaction between negatively charged CdTe and positively charged PDAC. The composition of the CdTe nanoparticle multilayer films was analyzed by X-ray photoelectron spectroscopy (XPS) combined with optical absorbance and luminescence measurements. It was experimentally observed that Cd-thiol complexes on the surface of the CdTe nanoparticles provide the crucial chemical passivation responsible for the high photoluminescence (PL) efficiency of the CdTe particles. The high PL efficiency and high stability of CdTe particles corresponded to the particles with the high surface coverage with Cd-thiol complexes. Moreover, XPS data indicated the surface coverage with Cd-thiol complexes could be increased around the CdTe particle by either reflux or adjusting the pH of resulted CdTe colloidal suspension, which was consistent with the results from optical absorbance and luminescence spectra. It appeared that the popular method of constructing multilayer films could be used as a tool to characterize the surface composition of nanometer-sized particles.