An electroactive microwell array for trapping and lysing single-bacterial cells

Abstract

Interest in single-cell analysis has increased because it allows to understand cell metabolism and characterize disease states, cellular adaptation to environmental changes, cell cycles, etc. Here, the authors propose a device to electrically trap and lyse single-bacterial cells in an array format for high-throughput single-cell analysis. The applied electric field is highly deformed and concentrated toward the inside of the microwell structures patterned on the planar electrode. This configuration effectively generates dielectrophoretic force to attract a single cell per well. The microwell has a comparable size to the target bacterial cell making it possible to trap single cells by physically excluding additional cells. Inducing highly concentrated electric potential on the cell membrane can also effectively lyse the trapped single-bacterial cells. The feasibility of the authors' approach was demonstrated by trapping and lysing Escherichia coli cells at the single-cell level. The present microwell array can be used as a basic tool for individual bacterial cell analysis.

Design and Control of Synthetic Biological Systems

“…To achieve the controllability of complex genetic circuits, genetic parts such as riboswitches should be developed. Technologies such as the electroactive microwell array [31] are also required to assess more states of cells, for measurability. Furthermore, for systematic control, a linear approximation of a mathematical model of cells with a synthetic circuit, such as piecewise affine approximation [32], would be required, because a non-linear system is difficult to control systematically.…”

A Brief Review of the Biophysical Hallmarks of Metastatic Cancer Cells

“…Recent advances in microfluidics technologies have opened the door to cancer biophysics and the capture and downstream analyses of circulating tumor cells (Becker, 2009; Beebe et al, 2002; Bhagat et al, 2010; Gossett et al, 2012; Haeberle and Zengerle, 2007; Hansen and Quake, 2003; Kim et al, 2011; Qin, 2012; Qin et al, 2008). A number of groups have published microfluidic approaches for screening the biophysical properties of suspended cells.…”

A generic and label free method based on dielectrophoresis for the continuous separation of microorganism from whole blood samples

“…More recently, other methods have emerged, that can separate cells according to differences in their physical parameters: hydrodynamic sorting [12], lateral displacement filtration [13], magnetophoresis [14], electrophoresis [15], acoustophoresis [16] or optical forces [17], AC electrokinetic techniques [18]. In particular, the use of dielectrophoresis forces, electrorotation, and travelling wave dielectrophoresis showed to be quite efficient for cell trapping [19][20][21], cell separation [22][23][24]and cell characterization [25,26]. These label-free and non-contact techniques offer the advantage of being minimally invasive and less sensitive to the inherent pitfalls of surface-mediated capture.…”