Most common methods of cellular analysis employ the top-down approach (investigating proteomics or genomics directly), thereby destroying the cell, which does not allow the possibility of using the same cell to correlate genomics with functional assays. Herein we describe an approach for single-cell tools that serve as a bottom-up approach. Our technology allows functional phenotyping to be conducted by observing the cytotoxicity of cells and then probe the underlying biology. We have developed a droplet microfluidic device capable of trapping droplets in the array and releasing the droplet of interest selectively using microvalves. Each droplet in the array encapsulates natural killer cells (NK cells) and tumour cells for real-time monitoring of burst kinetics and spatial coordination during killing by single NK cells. Finally, we use the microvalve actuation to selectively release droplets with the desired functional phenotype such as for fast and serial killing of target tumour cells by NK cells. From this perspective, our device allows for investigating first interactions and real-time monitoring of kinetics and later cell recovery on demand for single-cell omic analysis such as single-cell RNA sequencing (scRNA), which to date, is primarily based on in-depth analyses of the entire transcriptome of a relatively low number of cells.

中文翻译：

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使用微阀进行功能时间分析和按需细胞恢复的液滴微流体：在癌症免疫疗法中的应用

最常见的细胞分析方法采用自上而下的方法（直接研究蛋白质组学或基因组学），从而破坏细胞，这不允许使用相同的细胞将基因组学与功能测定关联起来。在这里，我们描述了一种作为自下而上方法的单细胞工具方法。我们的技术允许通过观察细胞的细胞毒性来进行功能表型分析，然后探究潜在的生物学。我们开发了一种液滴微流体装置，能够捕获阵列中的液滴并使用微阀选择性地释放感兴趣的液滴。阵列中的每个液滴都封装了自然杀伤细胞（NK 细胞）和肿瘤细胞，用于实时监测单个 NK 细胞杀伤过程中的爆发动力学和空间协调。最后，我们使用微阀驱动选择性地释放具有所需功能表型的液滴，例如 NK 细胞快速、连续地杀死目标肿瘤细胞。从这个角度来看，我们的设备可以根据单细胞组学分析（例如单细胞 RNA 测序 (scRNA)）的需要来研究首次相互作用和动力学实时监测以及随后的细胞恢复，迄今为止，单细胞组学分析主要基于-对相对较少数量的细胞的整个转录组进行深入分析。