In this study, a useful method was developed to fabricate array patterns of microparticles not on electrode surfaces, but on arbitrary surfaces, using negative-dielectrophoresis (n-DEP). First, electrodes were designed and electric field simulations were performed to manipulate microparticles toward target areas. Based on the simulation results, multilayered array and grid (MLAG) electrodes, consisting of array electrodes surrounded by insulated regions and a grid electrode, were fabricated for the formation of localized, non-uniform electric fields. The MLAG electrode was mounted to a target substrate in a face-to-face configuration with a spacer. When an AC voltage (4.60 V-rms and 1 MHz) was applied to the MLAG electrode, array patterns of 6 and 20 mu m diameter microparticles were rapidly fabricated on the target substrate with ease. The results suggest that MLAG electrodes can be widely applied for the fabrication of biochips including cell arrays. Biotechnol. Bioeng. 2009;104: 709-718. (C) 2009 Wiley Periodicals, Inc.