Literature DB >> 27528072

Parallelized cytoindentation using convex micropatterned surfaces.

Bojing Jia1,2, Tse-Luen Wee2,3,4, Colton G Boudreau3, Daniel J Berard1, Adiel Mallik5,6,7, David Juncker5,6,7, Claire M Brown3,4, Sabrina R Leslie1.   

Abstract

Here we present a high-throughput, parallelized cytoindentor for local compression of live cells. The cytoindentor uses convex lens-induced confinement (CLiC) to indent micrometer-sized areas in single cells and/or populations of cells with submicron precision. This is accomplished using micropatterned poly(dimethylsiloxane) (PDMS) films that are adhered to a convex lens to create arrays of extrusions referred to here as "posts." These posts caused local deformation of subcellular regions without any evidence of cell lysis upon CLiC indentation. Our micropost arrays were also functionalized with glycoproteins, such as fibronectin, to both pull and compress cells under customized confinement geometries. Measurements of Chinese hamster ovary (CHO-K1) cell migration trajectories and oxidative stress showed that the CLiC device did not damage or significantly stress the cells. Our novel tool opens a new area of investigation for visualizing mechanobiology and mechanochemistry within living cells, and the high-throughput nature of the technique will streamline investigations as current tools for mechanically probing material properties and molecular dynamics within cells, such as traditional cytoindentors and atomic force microscopy (AFM), are typically restricted to single-cell manipulation.

Entities:  

Keywords:  cell loading; confocal microscopy; convex lens-induced confinement; cytoindentation; microfabrication

Mesh:

Substances:

Year:  2016        PMID: 27528072     DOI: 10.2144/000114436

Source DB:  PubMed          Journal:  Biotechniques        ISSN: 0736-6205            Impact factor:   1.993


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