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Diamagnetic droplet microfluidics applied to single-cell sorting

机译:抗磁性液滴微流控技术应用于单细胞分选

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The heterogeneity of diseased tissue causes major challenges in the detection and treatment of disease. Such challenges have motivated the development of tools for single-cell isolation and analysis. However, many cell isolation methods in microfluidics rely on the use of cell-labeling steps or expose cells to potentially harmful forces. Here, we present a microfluidic method for label-free control of cell-encapsulating biocompatible droplets using negative magnetophoresis. Our system is distinguished from previous microfluidic diamagnetic sorting approaches by the encapsulation of the cells inside droplets, which isolates the cells from the magnetic continuous phase. The droplet phase is comprised of cells suspended in their growth culture medium, and all of the magnetic content is contained in the oil-based continuous phase. At a flow-focusing junction, empty droplets and cell-encapsulating droplets are both generated and surrounded by the magnetic continuous phase. Cell encapsulation produces a size distinction between empty droplets and cell-encapsulated droplets. Through the application of a permanent magnet to the system, diamagnetic size-based sorting of empty droplets from cell-encapsulated droplets is achieved with a purity of ~84% in a single pass. Additionally, since the encapsulated cells are completely isolated from the magnetic content in the continuous phase, 88% cell-viability is observed after a two-hour incubation period. If combined with a single-cell assay, this system can provide label-free isolation of viable cells at a high purity for subsequent downstream analysis.
机译:患病组织的异质性给疾病的检测和治疗带来了重大挑战。这些挑战激发了单细胞分离和分析工具的开发。然而,微流控中的许多细胞分离方法都依赖于使用细胞标记步骤或使细胞暴露于潜在的有害作用力。在这里,我们提出了一种使用负磁泳的无标记控制细胞包裹生物相容性液滴的微流控方法。我们的系统与以前的微流体抗磁分选方法不同,其封装方式是将液滴封装在液滴内部,从而将细胞与磁性连续相隔离。液滴相由悬浮在其生长培养基中的细胞组成,所有磁性成分都包含在油基连续相中。在聚流结点处,空的液滴和细胞包裹的液滴都被生成并被磁性连续相包围。细胞包囊在空液滴和细胞包囊的液滴之间产生大小差异。通过将永磁体应用于该系统,单次通过基于抗磁尺寸的细胞包封液滴对空液滴进行分选,纯度约为84%。另外,由于被包封的细胞在连续相中与磁性成分完全隔离,因此在两个小时的孵育期后观察到88%的细胞活力。如果与单细胞分析结合使用,该系统可以提供无标记的高纯度活细胞分离,用于后续的下游分析。

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