Cell–cell interactions are essential for the proper functioning of multicellular organisms. For example, T cells interact with antigen-presenting cells (APCs) through specific T-cell receptor (TCR)–antigen interactions during an immune response. Fluorescence-activated droplet sorting (FADS) is a high-throughput technique for efficiently screening cellular interaction events. Unfortunately, current droplet sorting instruments have significant limitations, most notably related to analytical throughput and complex operation. In contrast, commercial fluorescence-activated cell sorters offer superior speed, sensitivity, and multiplexing capabilities, although their use as droplet sorters is poorly defined and underutilized. Herein, we present a universally applicable and simple-to-implement workflow for generating double emulsions and performing multicolor cell sorting using a commercial FACS instrument. This workflow achieves a double emulsion detection rate exceeding 90%, enabling multicellular encapsulation and high-throughput immune cell activation sorting for the first time. We anticipate that the presented droplet sorting strategy will benefit cell biology laboratories by providing access to an advanced microfluidic toolbox with minimal effort and cost investment.

中文翻译：

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用于多色荧光激活细胞分选的稳健双乳液


细胞与细胞的相互作用对于多细胞生物的正常运作至关重要。例如，在免疫反应过程中，T 细胞通过特定的 T 细胞受体 (TCR)-抗原相互作用与抗原呈递细胞 (APC) 相互作用。荧光激活液滴分选 (FADS) 是一种有效筛选细胞相互作用事件的高通量技术。不幸的是，当前的液滴分选仪器具有显着的局限性，最明显的是与分析通量和复杂的操作有关。相比之下，商业荧光激活细胞分选仪提供卓越的速度、灵敏度和多重功能，尽管它们作为液滴分选仪的用途尚不明确且未得到充分利用。在此，我们提出了一种普遍适用且易于实施的工作流程，用于生成双乳液并使用商用 FACS 仪器进行多色细胞分选。该工作流程实现了超过90%的双乳检测率，首次实现了多细胞封装和高通量免疫细胞激活分选。我们预计，所提出的液滴分选策略将通过以最少的努力和成本投资提供先进的微流体工具箱，从而使细胞生物学实验室受益。