Abstract
The electroporation system can serve as a tool for the intracellular delivery of foreign cargos. However, this technique is presently limited by the inaccurate electric field applied to the single cells and lack of a real-time electroporation metrics subsystem. Here, we reported a microfluidic system for precise and rapid single-cell electroporation and simultaneous impedance monitoring in a constriction microchannel. When single cells (A549) were continuously passing through the constriction microchannel, a localized high electric field was applied on the cell membrane, which resulted in highly efficient (up to 96.6%) electroporation. During a single cell entering the constriction channel, an abrupt impedance drop was noticed and demonstrated to be correlated with the occurrence of electroporation. Besides, while the cell was moving in the constriction channel, the stabilized impedance showed the capability to quantify the electroporation extent. The correspondence of the impedance variation and electroporation was validated by the intracellular delivery of the fluorescence indicator (propidium iodide). Based on the obtained results, this system is capable of precise control of electroporation and real-time, label-free impedance assessment, providing a potential tool for intracellular delivery and other biomedical applications.
Highlights
Intracellular delivery of foreign cargos is becoming a critical step in biomedical applications, ranging from gene editing to cell-based therapies [1]
We propose a microfluidic system for single-cell electroporation and real-time impedance monitoring without sacrificing the throughput
As the efficient charging of transmembrane voltage (TMV) could result in the occurrence of electroporation, the Finite Element Method (FEM) simulations of TMVs during cell-passing through the constriction channel were analyzed in COMSOL Multiphysics (COMSOL Multiphysics, COMSOL Inc., Burlington, MA, USA)
Summary
Intracellular delivery of foreign cargos is becoming a critical step in biomedical applications, ranging from gene editing to cell-based therapies [1]. The physical methods based on membrane disruption have the advantage of the cell/material independency for various applications [6,7,8,9,10,11]. In a conventional bulk electroporation (BEP) system, a mixture of cells and cargos is usually suspended in a cuvette full of conducting buffer, and two macro-electrodes with high voltage (~hundreds of volts) are applied [15]. These systems suffer from low delivery efficiency and intense cell damage due to the inhomogeneous, uncontrollable electric field across the cell suspension
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