一种简单的微流控方法,用于在机械和生化测定中选择、分离和操作单细胞。
A simple microfluidic method to select, isolate, and manipulate single-cells in mechanical and biochemical assays.
机构信息
Université de Mons, Laboratoire Interfaces & Fluides Complexes, Centre d'Innovation et de Recherche en Matériaux (CIRMAP), 20, Place du Parc, B-7000 Mons, Belgique.
出版信息
Lab Chip. 2010 Jun 7;10(11):1459-67. doi: 10.1039/c002257h. Epub 2010 Mar 24.
This article describes a simple and low-tech microfluidic method for single-cell experimentation, which permits cell selection without stress, cell manipulation with fine control, and passive self-exclusion of all undesired super-micronic particles. The method requires only conventional soft lithography microfabrication techniques and is applicable to any microfluidic single-cell circuitry. The principle relies on a bypass plugged in parallel with a single-cell assay device and collecting 97% of the flow rate. Cell selection into the single cell device is performed by moving the cell of interest back and forth in the vicinity of the junction between the bypass and the analysis circuitry. Cell navigation is finely controlled by hydrostatic pressure via centimetre-scale actuation of external macroscopic reservoirs connected to the device. We provide successful examples of biomechanical and biochemical assays on living human leukocytes passing through 4 mum wide capillaries. The blebbing process dynamics are monitored by conventional 24 fps videomicroscopy and subcellular cytoskeleton organization is imaged by on-chip immunostaining.
本文描述了一种简单的、低技术含量的微流控单细胞实验方法,该方法可以在不施加压力的情况下进行细胞选择,对细胞进行精细控制操作,并能被动地将所有不需要的超微颗粒排除在外。该方法仅需要常规的软光刻微加工技术,并且适用于任何微流控单细胞电路。该原理依赖于并联的旁路和单细胞分析装置,可收集 97%的流速。通过在旁路和分析电路之间的交界处附近来回移动感兴趣的细胞,将细胞选入单细胞装置中。通过外部宏观储液器以厘米级的幅度进行液压驱动,实现对细胞的精细导航控制,该储液器与装置相连。我们成功地对通过 4 微米宽毛细血管的活体人白细胞进行了生物力学和生物化学检测。通过常规的 24 fps 视频显微镜监测出芽过程的动力学,并通过芯片上免疫染色对细胞骨架的亚细胞组织进行成像。