This paper presents the fabrication and testing of a microfluidic device having a hydrophobic passive microvalve and a nano-hole array for releasing NGF (Nerve Growth Factors). This device was realized by processing a SOI (Silicon on Insulator) substrate. The microvalve was constructed in a cannel and driven by inlet pressure. It is possible to control NGF release precisely by opening and closing the microvalve. The nano-hole array was fabricated in a SiO_2 membrane of 500 nm in thickness using FIB (Focused Ion Beam) etching. Each nano-hole measures 500 nm in diameter, and is smaller than the diameter of an axon terminal. Experiments using a fluorescent solution showed that chemical release through the nano-holes was successfully controlled by opening and closing the microvalve. This result suggests that the fabricated device can axonal guidance by controlling release of NGF.