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Proc Natl Acad Sci U S A. 2017 Jul 11;114(28):7283-7288. doi: 10.1073/pnas.1705059114. Epub 2017 Jun 26.

Microfluidic guillotine for single-cell wound repair studies.

Author information

1
Department of Mechanical Engineering, Stanford University, Stanford, CA 94305.
2
Department of Biochemistry and Biophysics, University of California, San Francisco, CA 94143.
3
Department of Mechanical Engineering, Stanford University, Stanford, CA 94305; sindy@stanford.edu.

Abstract

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 Stentor coeruleus cells in a continuous-flow manner. Stentor 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 Stentor, enables time-course mechanistic studies impossible with current wounding methods.

KEYWORDS:

Stentor coeruleus; microfluidics; microguillotine; single cell; wound healing

PMID:
28652371
PMCID:
PMC5514750
[Available on 2018-01-11]
DOI:
10.1073/pnas.1705059114
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