Li Xuan, Lee Abraham P
Department of Biomedical Engineering, University of California, Irvine, CA, United States.
Department of Biomedical Engineering, University of California, Irvine, CA, United States; Department of Mechanical & Aerospace Engineering, University of California, Irvine, CA, United States.
Methods Cell Biol. 2018;148:35-50. doi: 10.1016/bs.mcb.2018.09.010. Epub 2018 Nov 5.
Single-cell analysis is of critical importance in revealing population heterogeneity, identifying minority sub-populations of interest, as well as discovering unique characteristics of individual cells. Microfluidic platforms work at the scale comparable to cell diameter and is suitable for single-cell manipulation. Here we present a microfluidic trapping array which is able to rapidly and deterministically trap single-cells in highly-packed microwells. This chapter first describes the design and fabrication protocols of the trapping array, and then presents its two representative applications: single-cell mRNA probing when integrated with a dielectrophoretic nanotweezer (DENT), and live-cell real-time imaging when combined with fluorescence lifetime imaging microscopy (FLIM). As the single-cell trapping efficiency is determined by the channel design instead of the flow rate, this trapping array can be coupled with different microfluidic sample processing units with different flow rates for various single-cell analyses.
单细胞分析对于揭示群体异质性、识别感兴趣的少数亚群以及发现单个细胞的独特特征至关重要。微流控平台在与细胞直径相当的尺度上工作，适用于单细胞操作。在此，我们展示了一种微流控捕获阵列，它能够在高度密集的微孔中快速且确定性地捕获单细胞。本章首先描述捕获阵列的设计和制造方案，然后介绍其两个代表性应用：与介电泳纳米镊子（DENT）集成时的单细胞mRNA探测，以及与荧光寿命成像显微镜（FLIM）结合时的活细胞实时成像。由于单细胞捕获效率由通道设计而非流速决定，该捕获阵列可与具有不同流速的不同微流控样品处理单元耦合，用于各种单细胞分析。
