Deterministic Cell Pairing with Simultaneous Microfluidic Merging and Sorting of Droplets

IF 5.4 2区 工程技术 Q1 BIOCHEMICAL RESEARCH METHODS
Lab on a Chip Pub Date : 2025-09-23 DOI:10.1039/d5lc00627a
Kevin Michael Joslin, Sophia Dateshidze, Seung Won Shin, Adam R Abate, Iain Clark
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引用次数: 0

Abstract

Cell–cell interactions drive immune activation, tissue repair, and stem cell fate, yet there are few methods that can create large numbers of pre-defined cell pairs to study cell crosstalk. Droplet microfluidics allows high-throughput compartmentalization of multiple cells, but random loading results in <1% of droplets containing the desired combinations. Here, we present Pair Isolation by Coalescence and Sorting (PICS), a microfluidic platform that can generate specific cell pairs through droplet merging and sorting (‘merge-sorting’). PICS detects target combinations using fluorescence and triggers simultaneous electrocoalescence and dielectrophoretic sorting. Using fluorescent dye–loaded droplets, we achieved 98.6% purity of merged and sorted droplets. In experiments using cells stained with three distinct dyes, >90% of desired cell pairs were recovered – compared to fewer than 1% when using random Poisson loading. To demonstrate the utility of PICS for extended co-culture studies, we merged cells in an alginate solution with calcium chloride droplets, producing monodisperse alginate hydrogels in which 93.3% of the beads contained target cell pairs that maintained viability over 18 hours. Compared to selective merger, this approach physically isolates desired droplets, eliminating unmerged contaminants and enabling cleaner downstream workflows. PICS allows off-chip pre-incubation of droplets before pairing, the merger of reagents for multi-step assays, and the rapid isolation of desired droplet pairs – capabilities not jointly accessible with existing approaches. In summary, PICS is a flexible platform to enrich specific combinations of droplets, cells, or particles for high-throughput studies of cell crosstalk.
微流体同时合并和分选的确定性细胞配对
细胞-细胞相互作用驱动免疫激活、组织修复和干细胞命运,然而很少有方法可以创建大量预定义的细胞对来研究细胞串扰。液滴微流体允许对多个细胞进行高通量区隔,但随机加载导致1%的液滴含有所需的组合。在这里,我们提出了通过聚结和分选对分离(PICS),这是一种微流体平台,可以通过液滴合并和分选(“合并分选”)产生特定的细胞对。PICS检测目标组合使用荧光和触发同步电聚结和介电泳分类。使用荧光染料负载的液滴,合并和分选的液滴纯度达到98.6%。在用三种不同染料染色细胞的实验中,90%的所需细胞对被回收,而使用随机泊松加载时,这一比例不到1%。为了证明PICS在延长共培养研究中的效用,我们将细胞与氯化钙液滴混合在海藻酸盐溶液中,产生单分散的海藻酸盐水凝胶,其中93.3%的珠含有目标细胞对,可保持18小时以上的活力。与选择性合并相比,这种方法在物理上隔离了所需的液滴,消除了未合并的污染物,实现了更清洁的下游工作流程。PICS允许在配对前对液滴进行片外预孵育,合并试剂进行多步骤分析,以及快速分离所需的液滴对-现有方法无法共同实现的功能。总之,PICS是一个灵活的平台,可以丰富液滴、细胞或颗粒的特定组合,用于细胞串扰的高通量研究。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Lab on a Chip
Lab on a Chip 工程技术-化学综合
CiteScore
11.10
自引率
8.20%
发文量
434
审稿时长
2.6 months
期刊介绍: Lab on a Chip is the premiere journal that publishes cutting-edge research in the field of miniaturization. By their very nature, microfluidic/nanofluidic/miniaturized systems are at the intersection of disciplines, spanning fundamental research to high-end application, which is reflected by the broad readership of the journal. Lab on a Chip publishes two types of papers on original research: full-length research papers and communications. Papers should demonstrate innovations, which can come from technical advancements or applications addressing pressing needs in globally important areas. The journal also publishes Comments, Reviews, and Perspectives.
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