Individual shells of the diatom Coscinodiscus were self-assembled into a rectangular array on a glass surface that possessed a polyetectrolyte multilayer patterned through inkjet printing. This patterned thin film possessed hierarchical order with nanostructure provided by the diatom biosilica. The process used two polyetectrolytes with opposite electric potentials to control the surface charge of the substrate. The fine features of the diatom frustules were perfectly preserved as a result of the mild conditions of the deposition process. This technique has the potential to enable large-scale device applications that harness the unique properties of functionalized diatom biosilica.