We present single-cell pairing of different types of cells with a rapid manipulation based on positive dielectrophoresis (p-DEP). The DEP device for the manipulation of cells consisted of a microfluidic channel with an upper indium tin oxide (ITO) electrode and a lower ITO electrode with 10,000 (100 × 100) microwells. The width (14 μm) and depth (25 μm) of the individual microwells restricted the size to two vertically aligned cells. Cells stained in blue and stained in green were continuously trapped in the microwells. Cells were paired within only 1 min and a pairing efficiency of 53% was achieved.