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Trapping cells on a stretchable microwell array for single-cell analysis

机译:在可伸缩微孔阵列上捕获细胞以进行单细胞分析

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摘要

There is a need for a technology that can be incorporated into routine laboratory procedures to obtain a continuous, quantitative, fluorescence-based measurement of the dynamic behaviors of numerous individual living cells in parallel, while allowing other manipulations, such as staining, rinsing, and even retrieval of targeted cells. Here, we report a simple, low-cost microarray platform that can trap cells for dynamic single-cell analysis of mammalian cells. The elasticity of polydimethylsiloxane (PDMS) was utilized to trap tens of thousands of cells on an array. The PDMS microwell array was stretched by a tube through which cells were loaded on the array. Cells were trapped on the array by removal of the tube and relaxation of the PDMS. Once that was accomplished, the cells remained trapped on the array without continuous application of an external force and permitted subsequent manipulations, such as staining, rinsing, imaging, and even isolation of targeted cells. We demonstrate the utility of this platform by multicolor analysis of trapped cells and monitoring in individual cells real-time calcium flux after exposure to the calcium ionophore ionomycin. Additionally, a proof of concept for target cell isolation was demonstrated by using a microneedle to locally deform the PDMS membrane in order to retrieve a particular cell from the array.
机译:需要一种可以结合到常规实验室程序中的技术,以获取连续的,定量的,基于荧光的,平行的众多单个活细胞动态行为的测量,同时允许进行其他操作,例如染色,漂洗和甚至取回目标细胞。在这里,我们报告了一个简单,低成本的微阵列平台,该平台可以捕获细胞以进行哺乳动物细胞的动态单细胞分析。聚二甲基硅氧烷(PDMS)的弹性可用来捕获阵列上成千上万个细胞。用管将PDMS微孔阵列拉伸,通过该管将细胞加载到阵列上。通过移开试管和放松PDMS,将细胞捕获在阵列上。完成此操作后,无需继续施加外力即可将细胞捕获在阵列上,并允许后续操作,例如染色,漂洗,成像,甚至分离目标细胞。我们通过对捕获的细胞进行多色分析并在暴露于钙离子载体离子霉素后监测单个细胞中的实时钙通量来证明该平台的实用性。另外,通过使用微针局部变形PDMS膜以从阵列中回收特定细胞,证明了靶细胞分离的概念证明。

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