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Although previously developed Dielectrophoresis activated cell sorters (DACSes) have achieved high separation efficiency, limitations still exist with respect to the low throughput. In this paper, therefore, a numerical analysis is presented for a novel DACS to achieve not only high efficiency but also high throughput. Additionally, experimental validation is performed based on the results of a numerical simulation. First, streamlines in the channel are investigated according to various flow rate ratios in each outlet. After cells are deflected by a sufficient amount of dielectrophoretic force, they arrive at the tip of the last electrode pair. Their movement is then determined by hydrodynamic force. Therefore, it is important to confirm whether or not the streamline at the tip of the last electrode pair connects to the target area. Based on the streamline analysis results, the particle trajectory under various AC electric fields is investigated using a three-dimensional analysis. Accordingly, an input voltage condition (7 Vp-p at 10 kHz) is selected to confirm the influence of dielectrophoretic force. Conclusively, optimal flow rates under the determined input voltage condition are selected as 10 μl/min and 20 μl/min in outlet A (target area) and outlet B (non-target area), respectively. In order to validate the feasibility of the selected flow rates, a separation test is performed employing a K562 cell-bone marrow cancer cell-as a target cell. In the experiment, the proposed DACS achieves a high throughput of 45000 cells/min as well as a separation efficiency of 96.5% and recovery rate of 47.25%. [ABSTRACT FROM AUTHOR]