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一种使用微流控静态液滴阵列进行单细胞分离和回收的简易操作方法。

An easy-to-operate method for single-cell isolation and retrieval using a microfluidic static droplet array.

机构信息

School of Biomedical Engineering, University of Technology Sydney, Sydney, NSW, 2007, Australia.

Institute for Biomedical Materials and Devices (IBMD), Faculty of Science, University of Technology Sydney, Sydney, NSW, 2007, Australia.

出版信息

Mikrochim Acta. 2021 Jul 6;188(8):242. doi: 10.1007/s00604-021-04897-9.

Abstract

In-depth study of cellular heterogeneity of rare cells (e.g. circulating tumour cells (CTCs) and circulating foetal cells (CFCs)) is greatly needed in disease management but has never been completely explored due to the current technological limitations. We have developed a retrieval method for single-cell detection using a static droplet array (SDA) device through liquid segmentation with almost no sample loss. We explored the potential of using SDA for low sample input and retrieving the cells of interest using everyday laboratory equipment for downstream molecular analysis. This single-cell isolation and retrieval method is low-cost, rapid and provides a solution to the remaining challenge for single rare cell detection. The entire process takes less than 15 min, is easy to fabricate and allows for on-chip analysis of cells in nanolitre droplets and retrieval of desired droplets. To validate the applicability of our device and method, we mimicked detection of single CTCs by isolating and retrieving single cells and perform real-time PCR on their mRNA contents.

摘要

深入研究稀有细胞(例如循环肿瘤细胞(CTCs)和循环胎儿细胞(CFCs))的细胞异质性在疾病管理中非常重要,但由于当前技术限制,从未得到完全探索。我们开发了一种使用静态液滴阵列(SDA)设备通过液段化进行单细胞检测的检索方法,几乎没有样品损失。我们探索了使用 SDA 进行低样品输入的潜力,并使用日常实验室设备检索感兴趣的细胞,用于下游分子分析。这种单细胞分离和检索方法成本低、速度快,为单细胞稀有细胞检测的剩余挑战提供了一种解决方案。整个过程耗时不到 15 分钟,易于制造,并允许在纳升液滴中分析细胞和检索所需的液滴。为了验证我们的设备和方法的适用性,我们通过分离和检索单个细胞模拟了对单个 CTC 的检测,并对其 mRNA 含量进行了实时 PCR。

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