A microwell array device with integrated microfluidic components for enhanced single-cell analysis
A microwell array device with integrated microfluidic components for enhanced single-cell analysis
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DOI：
10.1002/elps.200900572
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发表时间：
2009-12-01
期刊：
影响因子：
2.9
通讯作者：
Andersson-Svahn, Helene
中科院分区：
文献类型：
--
作者：
Lindstrom, Sara;Mori, Kiichiroh;Andersson-Svahn, Helene
越来越多的人意识到细胞异质性在许多生物学和医学背景下的重要性，这促使人们对允许单细胞分析而不是传统的批量分析（从大量细胞中提供感兴趣的变量的平均值）的系统越来越感兴趣。最近，我们提出了一种用于长期、高通量单细胞分析的微孔芯片。该芯片已被证明在筛选个体癌症和干细胞的蛋白质/基因标记等方面很有用。然而，井中的液体只能通过手动冲洗芯片或其部分来添加或改变。这种方法有几个众所周知的缺点——包括交叉污染的风险、死体积大和费力——但是以前很少有人尝试将液体冲洗/切换通道集成到“即用型”单细胞分析系统中。在这里，我们设计了一个微孔系统（使用流动模拟），用于单细胞分析，集成了微流体组件（微通道，磁驱动微泵和储液器），为细胞培养孔提供试剂或冲洗，从而促进控制，定向液体处理。它可以完全独立使用，因为油管不是必需的。通过在微孔中培养内皮细胞，并成功应用活细胞钙黄蛋白AM染色，证明了集成系统的实用性。这种将高密度微孔芯片与微流控元件相结合的系统在许多筛选应用中具有巨大的潜力，例如探索单个细胞之间药物反应的重要但通常未被检测到的异质性。
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.