Microfluidic on-demand droplet generation, storage, retrieval, and merging for single-cell pairing
Microfluidic on-demand droplet generation, storage, retrieval, and merging for single-cell pairing
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DOI：
10.1039/c8lc01178h
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发表时间：
2019-02-07
期刊：
影响因子：
6.1
通讯作者：
DeVoe, Don L.
中科院分区：
文献类型：
--
作者：
Babahosseini, Hesam;Misteli, Tom;DeVoe, Don L.
描述了一种多功能微流体平台，其在单个集成微流体装置中组合了按需水相液滴产生、多液滴存储和选自存储库的液滴的受控合并。该技术的一个独特方面是一种微流体捕集器设计，其包括液滴捕集室和与微阀集成的侧向旁路通道，该微阀支持在宽范围的单个液滴尺寸上捕获和合并多个液滴。包括微流体捕集器阵列的存储单元以先进先出方式操作，允许存储在库内的液滴在顺序地将所选择的液滴递送到下游合并区之前被分析，同时将其他液滴分流为废物。的微流体陷阱的性能进行了研究的旁路/室流体动力学阻力比，微室的几何形状，捕获的液滴体积，和整体流速的变化。然后，利用集成的微流体平台来证明需要以单细胞分辨率分离限定的细胞群体的基于细胞的测定所必需的操作步骤，包括将单个细胞包封在水相液滴载体内，筛选或孵育固定化的细胞包封的液滴，以及通过顺序液滴合并过程产生单个细胞的受控组合。除了其用于细胞分析的实用性之外，所提出的平台还代表了一种用于在广泛的基于液滴的微流体应用中使用的稳健液滴生成、存储和合并的通用方法。
A multifunctional microfluidic platform combining on-demand aqueous-phase droplet generation, multi-droplet storage, and controlled merging of droplets selected from a storage library in a single integrated microfluidic device is described. A unique aspect of the technology is a microfluidic trap design comprising a droplet trap chamber and lateral bypass channels integrated with a microvalve that supports the capture and merger of multiple droplets over a wide range of individual droplet sizes. A storage unit comprising an array of microfluidic traps operates in a first-in first-out manner, allowing droplets stored within the library to be analyzed before sequentially delivering selected droplets to a downstream merging zone, while shunting other droplets to waste. Performance of the microfluidic trap is investigated for variations in bypass/chamber hydrodynamic resistance ratio, micro-chamber geometry, trapped droplet volume, and overall flow rate. The integrated microfluidic platform is then utilized to demonstrate the operational steps necessary for cell-based assays requiring the isolation of defined cell populations with single cell resolution, including encapsulation of individual cells within an aqueous-phase droplet carrier, screening or incubation of the immobilized cell-encapsulated droplets, and generation of controlled combinations of individual cells through the sequential droplet merging process. Beyond its utility for cell analysis, the presented platform represents a versatile approach to robust droplet generation, storage, and merging for use in a wide range of droplet-based microfluidics applications.