Huang Xiaomin, Xing Xiaoxing, Ng Chun Ning, Yobas Levent
Department of Electronic and Computer Engineering, The Hong Kong University of Science and Technology, Clear Water Bay, Kowloon, Hong Kong, China.
College of Information Science and Technology, Beijing University of Chemical Technology, Beijing 100029, China.
Micromachines (Basel). 2019 Jul 19;10(7):488. doi: 10.3390/mi10070488.
Highly localized (point) constrictions featuring a round geometry with ultra-sharp edges in silicon have been demonstrated for the reagent-free continuous-flow rapid mechanical lysis of mammalian cells on a single-cell basis. Silicon point constrictions, robust structures formed by a single-step dry etching process, are arranged in a cascade along microfluidic channels and can effectively rupture cells delivered in a pressure-driven flow. The influence of the constriction size and count on the lysis performance is presented for fibroblasts in reference to total protein, DNA, and intact nuclei levels in the lysates evaluated by biochemical and fluoremetric assays and flow-cytometric analyses. Protein and DNA levels obtained from an eight-constriction treatment match or surpass those from a chemical method. More importantly, many intact nuclei are found in the lysates with a relatively high nuclei-isolation efficiency from a four-constriction treatment. Point constrictions and their role in rapid reagent-free disruption of the plasma membrane could have implications for integrated sample preparation in future lab-on-a-chip systems.
在硅中已证明存在高度局部化（点状）的缩颈结构，其具有圆形几何形状且边缘超锋利，可用于在单细胞基础上对哺乳动物细胞进行无试剂连续流快速机械裂解。硅点状缩颈是通过单步干法蚀刻工艺形成的坚固结构，沿微流控通道呈级联排列，能够有效裂解在压力驱动流中输送的细胞。针对成纤维细胞，通过生化和荧光测定以及流式细胞分析评估裂解物中的总蛋白、DNA和完整细胞核水平，呈现了缩颈尺寸和数量对裂解性能的影响。八缩颈处理获得的蛋白质和DNA水平与化学方法相当或更高。更重要的是，在四缩颈处理的裂解物中发现了许多完整细胞核，且细胞核分离效率相对较高。点状缩颈及其在无试剂快速破坏质膜中的作用可能会对未来的芯片实验室系统中的集成样品制备产生影响。
