In this work, we have implemented a new microfluidic chip which can capture and isolate single-cells in a microchamber array for various single-cell assays. Proposed microfluidic chip consists of two PDMS (Polydimethylsiloxane) layers: a microfluidic channel layer for injecting cells and reagents and an air channel for pneumatic actuation. Each microfluidic chip has 27 microchambers for single-cell culture and analysis. In order to verify our single-cell capturing scheme, we tested the fabricated chip with 15 mum-diameter polystyrene microbeads. We successfully trapped the microbeads in each the capture site with a capture rate higher than 80% and loaded them into the corresponding chambers. After loading, the chambers were isolated from one another by pneumatic actuation. The isolation was confirmed by flowing blue ink into the main flow channel.