This paper presents a novel device for the dielectrophoretic manipulation of particles and cells. A two-level isotropic etch of a glass substrate was used to create three-dimensional ridge-like structures in micrometer-sized channels. Due to the insulating properties of glass, locally patterned regions of nonuniform electric field form near the ridges when a dc field is applied along the channel. The ridges are designed using the method of faceted prisms, such that substantially uniform fields are produced on each side of the faceted interfaces that form each ridge. The dielectrophoretic force that results from the electric field gradient near the ridges is used to affect particle motion parallel to the ridges in the absence of a bulk pressure-driven flow. Trapping and deflection of particles and continuous concentration and separation of Bacillus subtilis from a two-component sample mixture are demonstrated. The flow of B. subtilis is restricted to a selected channel of a planar, multichannel device as a result of negative dielectrophoresis arising from the presence of the insulating ridges when the applied electric field exceeds a threshold of 30 V/mm. Dielectrophoresis has a negligible impact on 200-nm-diameter polystyrene particles under the same conditions.