Multi-target pathogen detection using heterogeneous medical samples require continuous filtering, sorting, and trapping of debris, bioparticles, and immunocolloids within a diagnostic chip. We present an integrated AC dielectrophoretic ͑DEP͒ microfluidic platform based on planar electrodes that form three-dimensional ͑3D͒ DEP gates. This platform can continuously perform these tasks with a throughput of 3 L / min. Mixtures of latex particles, Escherichia coli Nissle, Lactobacillus, and Candida albicans are sorted and concentrated by these 3D DEP gates. Surface enhanced Raman scattering is used as an on-chip detection method on the concentrated bacteria. A processing rate of 500 bacteria was estimated when 100 l of a heterogeneous colony of 10 7 colony forming units /ml was processed in a single pass within 30 min.