We describe a new, simple process for fabricating transparent quadrupolar electrode arrays enabling large-scale particle assembly by means of dielectrophoresis. In the first step, interdigitated electrode arrays are made by chemical wet etching of indium tin oxide (ITO). Then, the transition from a bipolar to a quadrupolar electrode arrangement is obtained by covering the electrode surface with a thin poly(dimethylsiloxane) (PDMS) film acting as an electrical insulation layer in which selective openings are formed using bond-detach lithography. The PDMS insulating layer thickness was optimized and controlled by adjusting experimental parameters such as the PDMS viscosity (modulated by the addition of heptane) and the PDMS spin-coating velocity. The insulating character of the PDMS membrane was successfully demonstrated by performing a dielectrophoretic assembly of polystyrene particles using interdigitated electrodes with and without a PDMS layer. The results show that the patterned PDMS film functions properly as an electrical insulation layer and allows the reconfiguration of the electric field cartography. Electric field simulations were performed in both configurations to predict the dielectrophoretic behavior of the particles. The simulation results are in perfect agreement with experiments, in which we demonstrated the formation of concentrated clusters of polystyrene particles and living cells of regular size and shape.