Combined Dielectric-Optical Characterization of Single Cells Using Dielectrophoresis-Imaging Flow Cytometry.

In this paper, we present a microfluidic flow cytometer for simultaneous imaging and dielectric characterization of individual biological cells within a flow. Utilizing a combination of dielectrophoresis (DEP) and high-speed imaging, this system offers a dual-modality approach to analyze both cell morphology and dielectric properties, enhancing the ability to analyze, characterize, and discriminate cells in a heterogeneous population. A high-speed camera is used to capture images of and track multiple cells in real-time as they flow through a microfluidic channel. A wide channel is used, enabling analysis of many cells in parallel. A coplanar electrode array perpendicular to cell flow is incorporated at the bottom of the channel to perform dielectrophoresis-based dielectric characterization. A frequency-dependent voltage applied to the array produces a non-uniform electric field, translating cells to higher or lower velocity depending on their dielectric polarizability. In this paper, we demonstrate how cell size, obtained by optical imaging, and DEP response, obtained by particle tracking, can be used to discriminate viable and non-viable Chinese hamster ovary cells in a heterogeneous cell culture. Multiphysics electrostatic-fluid dynamics simulation is used to develop a relationship between cell incoming velocity, differential velocity, size, and the cell's polarizability, which can subsequently be used to evaluate its physiological state. Measurement of a mixture of polystyrene microspheres is used to evaluate the accuracy of the cytometer.