Analytical Sciences

Development of a Microfluidic Platform for Single-cell Secretion Analysis Using a Direct Photoactive Cell-attaching Method

Kihoon JANG,*1 Hong Trang Thi NGO,*1 Yo TANAKA,*1,*2,*3 Yan XU,*4 Kazuma MAWATARI,*1,*2 and Takehiko KITAMORI*1,*2

*1 Department of Applied Chemistry, Graduate School of Engineering, The University of Tokyo, 7-3-1 Hongo, Bunkyo, Tokyo 113-8656, Japan
*2 Japan Science and Technology Agency (JST), 7-3-1 Sanbacho, Chiyoda, Tokyo 102-0075, Japan
*3 Quantitative Biology Center (QBiC), RIKEN, 2-2-3 Minatojima-Minamimachi, Chuo, Kobe, Hyogo 650-0047, Japan
*4 Nanoscience and Nanotechnology Research Center, Research Organization for 21st Century, Osaka Prefecture University, 1-1 Gakuen, Naka, Osaka 599-8570, Japan

A precise understanding of individual cellular processes is essential to meet the expectations of most advanced cell biology. Therefore single-cell analysis is considered to be one of possible approach to overcome any misleading of cell characteristics by averaging large groups of cells in bulk conditions. In the present work, we modified a newly designed microchip for single-cell analysis and regulated the cell-adhesive area inside a cell-chamber of the microfluidic system. By using surface-modification techniques involving a silanization compound, a photo-labile linker and the 2-methacryloyloxyethyl phosphorylcholine (MPC) polymer were covalently bonded on the surface of a microchannel. The MPC polymer was utilized as a non-biofouling compound for inhibiting non-specific binding of the biological samples inside the microchannel, and was selectively removed by a photochemical reaction that controlled the cell attachment. To achieve the desired single-macrophage patterning and culture in the cell-chamber of the microchannel, the cell density and flow rate of the culture medium were optimized. We found that a cell density of 2.0 × 10⁶ cells/ml was the appropriate condition to introduce a single cell in each cell chamber. Furthermore, the macrophage was cultured in a small size of the cell chamber in a safe way for 5 h at a flow rate of 0.2 μl/min under the medium condition. This strategy can be a powerful tool for broadening new possibilities in studies of individual cellular processes in a dynamic microfluidic device.

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