Submicron-sized colloidal particles have been self-organized into patterned arrangements on a substrate using a novel technique. At the substrate, a polyelectrolyte multilayer film has been deposited onto a chemically patterned surface; subsequently, the polyelectrolyte surface is immersed in an aqueous colloidal suspension of bare SiO2 microspheres or functionalized polystyrene latex particles. The colloids self-organize at the surface, driven by the spatially varied electrostatic and secondary interactions between the colloid and the substrate. The polyelectrolyte platform provides a strong bond to the colloids, imparting mechanical robustness which enables postprocessing of the patterned assemblies. An important advantage to this approach is that the use of a polyelectrolyte multilayer platform opens up the possibility of introducing functionality into the underlying layers. We have demonstrated control over the density and selectivity of particle adsorption. Three mechanisms have been used to control adsorption: (i) pH of the colloid suspension, which determines the ionization of the uppermost surface of the polyelectrolyte multilayer; (ii) ionic strength of the suspension, which determines the extent of charge screening about the colloid and polyelectrolyte; and (iii) concentration of added surfactant, which causes charge screening and introduces hydrophobic interactions between the surfactant and polyelectrolyte. Finally, an energy adsorption model is presented.