We developed nanostructured gradient wrinkle surfaces to establish a one-step screening process towards optimal assembly of soft and hard colloidal particles (microgel systems and silica particles). Thereby, we simplify studies on the influence of wrinkle dimensions (wavelength, amplitude) on particle properties and their alignment. In a combinatorial experiment, we optimize particle assembly regarding the ratio of particle diameter vs. wrinkle wavelength and packing density and point out differences between soft and hard particles. The preparation of wrinkle gradients in oxidized top layers on elastic poly(dimethylsiloxane) (PDMS) substrates is based on a controlled wrinkling approach. Partial shielding of the substrate during plasma oxidation is crucial to obtain two-dimensional gradients with amplitudes ranging from 7 to 230 nm and wavelengths between 250 and 900 nm.