Microfludic Chips for High-throughput Single-cell Analysis with Hydrodynamic focusing

• 作 者: Xue-Feng Yin Chun-Xiu XU
• 机 构: Institute of Microanalytical Systems, Department of Chemistry, Zhejiang University,310027, Hangzhou,
• 出 处: 第一届国际微纳尺度生物医学分离和分析技术学术会议暨第六届全国微全分析学术会议
• 发表日期: 2010年6期

@@A microfluidic system for high-throughput single-cell analysis is described, with functional integration of continuous introduction of individual cells， rapid dynamic lysis, and capillary electrophoretic (CE) separation with laser induced fluorescence (LIF) detection. A cross microfluidic chip with two sheath-flow channels located on the both sides of the sampling channel was developed. The labeled cells were hydrodynamically focused by sheath-flow streams and sequentially introduced into the cross section by the hydrostatic pressure created by adjusting liquid levels in the reservoirs. Combined with the electric filed applied on the separation channel, the aligned cells were driven into the separation channel and rapid lysed within 33 ms at the entry of separation channel the by Triton X-100 added in the sheath-flow solution (Fig.1). The maximum rate for introduction of individual cells into the separation channel was about 150 cells /min. The introduction of sheath-flow streams also significantly reduced the concentration of phosphate-buffered saline (PBS) injected into the separation channel along with single cells, thus reducing Joule heating during electrophoresis. The microfluidic system was evaluated by analysis of GSH and ROS in single erythrocytes (Fig.2). A throughput of 38 samples/min was obtained. The proposed method is simple, easy to use and high-throughout, allows the analysis of cell populations of considerable sizes to generate the results with statistical significance.