阵列单液滴的捕获及单细胞分析

Encapsulation of Monodisperse Microdroplet Arrays and Single-Cell Analysis

通过流动聚焦型微液滴生成装置进行油包水液滴的制备,比较了3种不同的单液滴阵列捕获芯片对单液滴的捕获效率和操作性能,选取上下开口圆形结构获得满意的单液滴捕获。进一步利用该液滴生成和捕获一体式芯片进行了液滴中单细胞的包覆、单细胞的实时染色和细胞存活状态的检测。在优化的实验条件下制备得到了粒径为75μm的单分散液滴,液滴捕获芯片中的单液滴截留率为100%,单细胞包覆率约为33%。对捕获液滴中的单细胞进行实时染色结果表明,在5 min内获得了满意的细胞核实时染色,并通过细胞死活实时染色对细胞存活状态直接进行了表征。

The monodisperse water-in-oil microdroplets were prepared by a flow-focusing microfluidic chip. The capture effciency and operating performame of three different encapsulation structures were compared. A circular structure with opening up and down was selected for better encapsulation of microdroplet. Further, single cell encapsulation in microdroplet, cell staining and cell viability were carried out using this chip. Under the optimum conditions,monodisperse microdroplets with radius of 75 μm were prepared,the encapsulation of microdroplet in chip and single cell in each droplet were 100% and 33% , respectively. Also, staining results of single cell in capture droplets showed that the cell nucleus staining in microdroplets was accomplished within 5 min cell. The viability of cells in droplets was evaluated directly via live-dead staining in real time.