Patterning cells on a substrate is useful for cell assay and characterization. In this paper, a microchip employing negative dielectrophoresis was designed for efficient cell pattering. This chip consists of a 4-by-4 dot electrode array and the cell patterns are formed on a substrate lying on the microchip. To facilitate the microchip design, electric fields generated from the microchip were first simulated using software, and experiments were conducted to validate the performance of the microchip. Yeast cells suspending in the 6-aminohexanoic acid (AHA) solution were used in this study, and the effects of the cell medium, the voltage input and the voltage frequency were analyzed. The results confirm that cell patterns are successfully created with the microchip and this microchip offers an easy method to fabricate cell patterns on a substrate for characterization.