In this paper, we demonstrate a perfectly-ordered microbowl array with balanced dielectrophoresis (DEP) for a high-throughput single-cell analysis. In order to fabricate well-ordered microbowl array in a large area, we utilized three-dimensional diffuser lithography for photoresist mold and nickel electroplating technique for final microbowl structures on a silicon substrate. Single microbowl has six sharp apexes surrounding the microbowl perimeter. Each microbowl has a diameter of 10 μm, and a height of 9 μm, which can be controllable by patterns on mask and lithography conditions. To investigate feasibility for application to the microbowl array as a single-cell microarray, we used latex beads of 6.4 μm in an average diameter to be captured by dielectrophoretic force. The nickel microbowl array densely packed with a hexagonal geometry played as a bottom electrode, and an ITO-coated glass covered the nickel microbowl array as a top electrode while keeping a uniform gap between two electrodes. After injecting solution containing latex beads through the gap, we applied an AC signal (2 VPP, 1 MHz) between two electrodes to induce high electric field near the sharp apexes of the single microbowl. A negative DEP trap is formed at the center of the single microbowl with balanced DEP force from the six apexes. The experimental result shows that injected latex beads had been successfully and uniformly aligned and trapped at the microbowl array sustained by negative DEP.