Parallelized Cytoindentation Using Convex Micropatterned Surfaces

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.

Finite element simulation for the effect of loading rate on visco‐hyperelastic characterisation of soft materials by spherical nanoindentation

“…Each type has its own advantages and disadvantages. Cell indentation with a flat end tip is also known as cytoindentation [27]. Normally, the size of indentation tip is smaller than the cell.…”

Single-cell Analysis for Drug Development Using Convex Lens-Induced Confinement Imaging

“…This technique had been shown to be able to successfully confine and isolate diffusing [5] and interacting molecules [10][11][12] for long periods and improve fluorescent signal detection [7]. To date, CLiC has primarily been used to study materials, biomolecules and particles, with only limited information available regarding its suitability for live-cell investigations [13].…”

“…This device also holds great potential for routine use in fields such as cancer research, where single-cell variabilities and instabilities are especially important for developing effective therapies. While we focused on yeast cells as a model system in this work, previous studies performed on adherent Chinese hamster ovary (CHO-K1) cells demonstrated the feasibility of customizing the CLiC instrument with a cytoindentor, which allows precise indentation and imaging of live mammalian cells without significantly changing their oxidative stress or causing detectable impacts on cell fitness [13]. To manipulate cell adherence, the flow cells could be customized and coated with fibronectin as previously demonstrated [13].…”

“…While we focused on yeast cells as a model system in this work, previous studies performed on adherent Chinese hamster ovary (CHO-K1) cells demonstrated the feasibility of customizing the CLiC instrument with a cytoindentor, which allows precise indentation and imaging of live mammalian cells without significantly changing their oxidative stress or causing detectable impacts on cell fitness [13]. To manipulate cell adherence, the flow cells could be customized and coated with fibronectin as previously demonstrated [13]. Further optimization and validation studies will, however, be Reports needed for application on drug response and proliferation studies of mammalian cell lines.…”

Scattered Point Cloud Data Reconstruction Algorithm Based on Local Convexity

“…The demand for the precision of the point cloud data processing is getting higher and higher, and the point cloud data processing technology is developing continuously [4]. In the process of measurement, because there are no obvious geometric distribution in most cases and no obvious topological structure between data points with no organizational order, the whole point cloud presents a state of scattered and irregularities [5]. The reconstruction of the scattered point cloud data has become the key subject for researchers [6].…”