Department of Mechanical Engineering, Stanford University, Stanford, CA 94305.
Department of Biochemistry and Biophysics, University of California, San Francisco, CA 94143.
Proc Natl Acad Sci U S A. 2017 Jul 11;114(28):7283-7288. doi: 10.1073/pnas.1705059114. Epub 2017 Jun 26.
Wound repair is a key feature distinguishing living from nonliving matter. Single cells are increasingly recognized to be capable of healing wounds. The lack of reproducible, high-throughput wounding methods has hindered single-cell wound repair studies. This work describes a microfluidic guillotine for bisecting single cells in a continuous-flow manner. is used as a model due to its robust repair capacity and the ability to perform gene knockdown in a high-throughput manner. Local cutting dynamics reveals two regimes under which cells are bisected, one at low viscous stress where cells are cut with small membrane ruptures and high viability and one at high viscous stress where cells are cut with extended membrane ruptures and decreased viability. A cutting throughput up to 64 cells per minute-more than 200 times faster than current methods-is achieved. The method allows the generation of more than 100 cells in a synchronized stage of their repair process. This capacity, combined with high-throughput gene knockdown in , enables time-course mechanistic studies impossible with current wounding methods.
伤口修复是区分有生命和无生命物质的关键特征。越来越多的研究表明，单细胞具有修复伤口的能力。然而，由于缺乏可重复、高通量的创伤方法，单细胞伤口修复研究受到了阻碍。本工作描述了一种用于连续流动方式将单细胞二等分的微流控切刀。由于其强大的修复能力和高通量进行基因敲低的能力，我们选择 作为模型。局部切割动力学揭示了细胞被二等分的两种状态，一种是在低粘性应力下，细胞发生小的膜破裂，细胞活力较高，另一种是在高粘性应力下，细胞发生延伸的膜破裂，细胞活力降低。切割通量高达每分钟 64 个细胞——比当前方法快 200 多倍。该方法允许在修复过程的同步阶段产生 100 多个细胞。这种能力，结合在 中的高通量基因敲低，使我们能够进行当前创伤方法无法进行的时间进程机制研究。
