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使用带有双微阀的微流控芯片对单细胞进行动态筛选和打印。

Dynamic screening and printing of single cells using a microfluidic chip with dual microvalves.

作者信息

Chen Chang, Xu Dong, Bai Siwei, Yu Zhihang, Zhu Yonggang, Xing Xiao, Chen Huaying

机构信息

School of Mechanical Engineering and Automation, Harbin Institute of Technology, Shenzhen, Shenzhen 518055, China.

出版信息

Lab Chip. 2020 Apr 7;20(7):1227-1237. doi: 10.1039/d0lc00040j. Epub 2020 Feb 26.

DOI:10.1039/d0lc00040j
PMID:32100799
Abstract

Inoculation of single cells into separate culture chambers is one of the key requirements in single-cell analysis. This paper reports an innovative microfluidic chip integrating two pneumatic microvalves to screen and print single cells onto a well plate. The upper and lower size limits of cells can be dynamically controlled by regulating the deformation of two adjacent microvalves. Numerical simulations were employed to systematically study the influence of membrane dimensions and pressure on the deflection of a valve. A mathematical model was then modified to predict the size of cells captured by a microvalve at various pressures. The membrane deflection was further studied using confocal imaging. The critical pressure trapping beads of various sizes was experimentally determined. These experiments validated the accuracy of both numerical simulations and the mathematical model. Furthermore, single beads and endothelial cells with the desired size range were screened using dual valves and printed onto well plates with 100% efficiency. Viability studies suggested that the screening process had no significant impact on cells. This device enables dynamic regulation of both the lower and the upper size limits of cells for printing. It has significant application potential in inoculating cells with desired sizes for various fields such as clonal expansion, monoclonality development and single-cell genomic studies.

摘要

将单细胞接种到单独的培养室是单细胞分析的关键要求之一。本文报道了一种集成了两个气动微阀的创新型微流控芯片,用于筛选单细胞并将其打印到微孔板上。通过调节两个相邻微阀的变形,可以动态控制细胞的大小上限和下限。采用数值模拟系统地研究了膜尺寸和压力对阀偏转的影响。然后修改了一个数学模型,以预测在不同压力下微阀捕获的细胞大小。使用共聚焦成像进一步研究了膜的偏转。通过实验确定了捕获各种尺寸珠子的临界压力。这些实验验证了数值模拟和数学模型的准确性。此外,使用双阀筛选出了具有所需尺寸范围的单个珠子和内皮细胞,并以100%的效率打印到微孔板上。活力研究表明,筛选过程对细胞没有显著影响。该装置能够动态调节用于打印的细胞大小的下限和上限。它在为克隆扩增、单克隆性发展和单细胞基因组研究等各个领域接种所需大小的细胞方面具有重要的应用潜力。

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