This article discusses a new, general approach for fabricating surfaces with precise positional control of chemical functionalities utilizing direct patterning of self-assembled (SA) organosilane monolayer films with lithographic exposure tools, including deep ultraviolet, x-ray, and e-beam sources. Lithographically patterned one- and two-component SA films have been used to selectively deposit or attach a wide variety of materials to surfaces, including catalysts, electroless metal films, proteins, cells, and organic moieties. Selectively metallized, patterned SA films have been employed to fabricate functioning Si metal-oxide-semiconductor field effect transistor test structures. The utility of patterned SA films for microelectronics, sensors, and other applications is discussed.