This study introduces a new generation of dielectrophoretic-based microfluidic device for the precise separation of multiple particle/cell types. The device features two sets of 3D electrodes, namely cylindrical and sidewall electrodes. The main channel of the device terminates with three outlets: one in the middle for particles that sense negative dielectrophoresis force and two others at the right and left sides for particles that sense positive dielectrophoresis force. To evaluate the device performance, we used red blood cells (RBCs), T-cells, U937-MC cells, and Clostridium difficile bacteria as our test subjects. Our results demonstrate that the proposed microfluidic device could accurately separate bioparticles in two steps, with sidewall electrodes of 200 µm proving optimal for efficient separation. Applying different voltages for each separation step, we found that the device performed most effectively at 6 Vp-p applied to the 3D electrodes, and at 20 Vp-p and 11 Vp-p applied to the sidewall electrodes for separating RBCs from bacteria and T-cells from U937-MC cells, respectively. Notably, the device's maximum electric fields remained below the cell electroporation threshold, and we achieved a separation efficiency of 95.5% for multi-type particle separation. Our findings proved the device's capacity for separating multiple particle types with high accuracy, without limitation for particle variety.