Division of Nanobiotechnology, School of Biotechnology, AlbaNova University Center, Royal Institute of Technology, Stockholm, Sweden.
Electrophoresis. 2009 Dec;30(24):4166-71. doi: 10.1002/elps.200900572.
Increasing awareness of the importance of cell heterogeneity in many biological and medical contexts is prompting increasing interest in systems that allow single-cell analysis rather than conventional bulk analysis (which provides average values for variables of interest from large numbers of cells). Recently, we presented a microwell chip for long-term, high-throughput single-cell analysis. The chip has proved to be useful for purposes such as screening individual cancer and stem cells for protein/gene markers. However, liquids in the wells can only be added or changed by manually rinsing the chip, or parts of it. This procedure has several well-known drawbacks--including risks of cross-contamination, large dead volumes and laboriousness--but there have been few previous attempts to integrate liquid rinsing/switching channels in "ready-to-use" systems for single-cell analysis. Here we present a microwell system designed (using flow simulations) for single-cell analysis with integrated microfluidic components (microchannels, magnetically driven micropumps and reservoirs) for supplying the cell culture wells with reagents, or rinsing, thus facilitating controlled, directed liquid handling. It can be used totally independently, since tubing is not essential. The practical utility of the integrated system has been demonstrated by culturing endothelial cells in the microwells, and successfully applying live-cell Calcein AM staining. Systems such as this combining high-density microwell chips with microfluidic components have great potential in numerous screening applications, such as exploring the important, but frequently undetected, heterogeneity in drug responses among individual cells.
在许多生物学和医学背景下，人们越来越意识到细胞异质性的重要性，这促使人们对能够进行单细胞分析的系统产生了越来越大的兴趣，而不是传统的批量分析（批量分析为大量细胞的感兴趣变量提供平均值）。最近，我们提出了一种用于长期、高通量单细胞分析的微井芯片。该芯片已被证明可用于筛选单个癌细胞和干细胞的蛋白质/基因标志物等目的。然而，微井中的液体只能通过手动冲洗芯片或其部分来添加或更换。该过程存在几个众所周知的缺点，包括交叉污染、死体积大和繁琐的风险，但以前很少有尝试将液体冲洗/切换通道集成到“即用型”单细胞分析系统中。在这里，我们提出了一种微井系统，该系统使用流动模拟设计用于单细胞分析，集成了微流控组件（微通道、磁驱动微泵和储液器），用于向细胞培养井供应试剂或冲洗液，从而便于进行受控的定向液体处理。由于不需要使用管道，因此该系统可以完全独立使用。通过在微井中培养内皮细胞并成功应用活细胞 Calcein AM 染色，证明了集成系统的实用性。这种将高密度微井芯片与微流控组件相结合的系统在许多筛选应用中具有巨大的潜力，例如探索单个细胞中药物反应的重要但经常未被发现的异质性。
