Biomolecule attachment and lateral micropatterning of biomolecular layers are essential techniques to provide in advanced biochemical and cell culture assays. For that purpose, we introduced a versatile, simple and robust method to functionalise standard polystyrene well plates. Free amino groups were generated on the polystyrene surface by low pressure ammonia plasma treatment. Subsequently, thin films of different maleic anhydride copolymers were covalently attached to the surfaces. The distinct physicochemical properties of the coupled maleic anhydride copolymers provided a broad range of possible attachment schemes of proteins and polysaccharides. Micrometer-sized lateral patterns of these functional coatings were created by plasma etching through silicon masks and subsequent chemical conversion of the etched areas using poly(ethylene glycol). The approach facilitates a wide variety of cell culture experiments allowing a combination of biomolecule coupling and micropatterning within the multi-well plate technology.