This paper presents the fabrication and testing of a microfluidic device consisting of a chamber for cell culture, a microhole array for chemical release, and a microchannel for sheathflow generation. The microholes of 2 μm in diameter were opened in the 100 nm thick SiN membrane of the culture chamber bottom using photolithography and CF_4 plasma etching. The microchannel having three inlets was located directly under the culture chamber. Experiments using a fluorescent solution showed that chemical release through the microholes was successfully controlled by regulating flow rates of three fluids which form the sheathflow. This result suggests that the fabricated device can be used for temporal-spatial control of localized chemical stimulation to a cultured cell.