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Article

Single-Cell Point Constrictions for Reagent-Free High-Throughput Mechanical Lysis and Intact Nuclei Isolation

1
Department of Electronic and Computer Engineering, The Hong Kong University of Science and Technology, Clear Water Bay, Kowloon, Hong Kong, China
2
College of Information Science and Technology, Beijing University of Chemical Technology, Beijing 100029, China
3
Division of Biomedical Engineering, The Hong Kong University of Science and Technology, Clear Water Bay, Kowloon, Hong Kong, China
*
Author to whom correspondence should be addressed.
Micromachines 2019, 10(7), 488; https://doi.org/10.3390/mi10070488
Received: 29 May 2019 / Revised: 15 July 2019 / Accepted: 16 July 2019 / Published: 19 July 2019
(This article belongs to the Special Issue 10th Anniversary of Micromachines)
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. View Full-Text
Keywords: cell lysis; constriction; DNA; protein; microfluidic; nuclei cell lysis; constriction; DNA; protein; microfluidic; nuclei
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MDPI and ACS Style

Huang, X.; Xing, X.; Ng, C.N.; Yobas, L. Single-Cell Point Constrictions for Reagent-Free High-Throughput Mechanical Lysis and Intact Nuclei Isolation. Micromachines 2019, 10, 488. https://doi.org/10.3390/mi10070488

AMA Style

Huang X, Xing X, Ng CN, Yobas L. Single-Cell Point Constrictions for Reagent-Free High-Throughput Mechanical Lysis and Intact Nuclei Isolation. Micromachines. 2019; 10(7):488. https://doi.org/10.3390/mi10070488

Chicago/Turabian Style

Huang, Xiaomin, Xiaoxing Xing, Chun Ning Ng, and Levent Yobas. 2019. "Single-Cell Point Constrictions for Reagent-Free High-Throughput Mechanical Lysis and Intact Nuclei Isolation" Micromachines 10, no. 7: 488. https://doi.org/10.3390/mi10070488

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