Controllable in-situ cell electroporation with cell positioning and impedance monitoring using micro electrode array

Guo, Xiaoliang; Zhu, Rong

doi:10.1038/srep31392

Cited by 48 publications

(42 citation statements)

References 24 publications

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“…The microchip consisted of a plurality of sextupole-electrode units patterned in an array. In each unit, there were a pair of center microelectrodes and quadrupole positioning electrodes [33][34][35] . The center microelectrodes were used to perform in situ cellular EP and real-time impedance monitoring.…”

Section: System and Microchipmentioning

confidence: 99%

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Single-cell individualized electroporation with real-time impedance monitoring using a microelectrode array chip

2020

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The ability to precisely deliver molecules into single cells while maintaining good cell viability is of great importance to applications in therapeutics, diagnostics, and drug delivery as it is an advancement toward the promise of personalized medicine. This paper reports a single-cell individualized electroporation method with real-time impedance monitoring to improve cell perforation efficiency and cell viability using a microelectrode array chip. The microchip contains a plurality of sextupole-electrode units patterned in an array, which are used to perform in situ electroporation and real-time impedance monitoring on single cells. The dynamic recovery processes of single cells under electroporation are tracked in real time via impedance measurement, which provide detailed transient cell states and facilitate understanding the whole recovery process at the level of single cells. We define single-cell impedance indicators to characterize cell perforation efficiency and cell viability, which are used to optimize electroporation. By applying the proposed electroporation method to different cell lines, including human cancer cell lines and normal human cell lines individually, optimum stimuli are determined for these cells, by which high transfection levels of enhanced green fluorescent protein (EGFP) plasmid into cells are achieved. The results validate the effectiveness of the proposed single-cell individualized electroporation/transfection method and demonstrate promising potential in applications of cell reprogramming, induced pluripotent stem cells, adoptive cell therapy, and intracellular drug delivery technology.

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Section: System and Microchipmentioning

confidence: 99%

“…We have developed a microelectrode array chip that integrates multiple functions of cell positioning, MEP, and impedance monitoring [33][34][35][36] . In this paper, we propose a single-cell individualized EP method via real-time impedance monitoring to balance perforation efficiency and cell viability using the microchip.…”

Section: Introductionmentioning

confidence: 99%

Single-cell individualized electroporation with real-time impedance monitoring using a microelectrode array chip

2020

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“…Another important application of electroporation is small particle delivery for drug loading [8], living cell staining or transfection [9], which is crucial for elucidating the biological roles/duties of different cellular components and organelles. Conventional methods of living cell staining rely on the proper characteristics (like reactivity and hydrophilicity) of staining molecules that allow them to pass through the membranes of living cells spontaneously [10].…”

Section: Electroporationmentioning

confidence: 99%

A Novel Electroporation System for Living Cell Staining and Membrane Dynamics Interrogation

et al. 2020

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A novel electroporation system was developed to introduce transient membrane pores to cells in a spatially and temporally controlled manner, allowing us to achieve fast electrotransfection and live cell staining as well as to systematically interrogate the dynamics of the cell membrane. Specifically, using this platform, we showed that both reversible and irreversible electroporation could be induced in the cell population, with nano-sized membrane pores in the former case being able to self-reseal in ~10 min. In addition, green fluorescent protein(GFP)-vinculin plasmid and 543 phalloidin have been delivered successively into fibroblast cells, which enables us to monitor the distinct roles of vinculin and F-actin in cell adhesion and migration as well as their possible interplay during these processes. Compared to conventional bulk electroporation and staining methods, the new system offers advantages such as low-voltage operation, cellular level manipulation and testing, fast and adjustable transfection/staining and real-time monitoring; the new system therefore could be useful in different biophysical studies in the future.

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“…Boukany et al utilized a micro-nano-micro-channel for the precise delivery of transfection agents to the trapped single cells [ 27 ]. In single-cell electroporation systems, complicated micro/nano-fluidic channels [ 26 , 27 , 28 , 29 ] or single-cell micromanipulation units [ 30 , 31 , 32 , 33 ] were usually required. In addition, most of these systems were combined with an optical detection tool for visualizing electroporation results [ 34 ].…”

Section: Introductionmentioning

confidence: 99%

“…Besides, the use of fluorescent dyes is unsuitable for subsequent assays of the treated cells. Recently, electrical detections of the current [ 26 , 36 ] and impedance [ 30 , 37 , 38 , 39 , 40 , 41 ] have been reported to be incorporated with the electroporation system. Compared to the traditional optical detections, electrical detections provide a label-free and instant strategy for electroporation assessment.…”

Section: Introductionmentioning

confidence: 99%

Single-Cell Electroporation with Real-Time Impedance Assessment Using a Constriction Microchannel

Luan

Zhang

et al. 2020

Micromachines

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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.

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Controllable in-situ cell electroporation with cell positioning and impedance monitoring using micro electrode array

Cited by 48 publications

References 24 publications

Single-cell individualized electroporation with real-time impedance monitoring using a microelectrode array chip

Single-cell individualized electroporation with real-time impedance monitoring using a microelectrode array chip

A Novel Electroporation System for Living Cell Staining and Membrane Dynamics Interrogation

Single-Cell Electroporation with Real-Time Impedance Assessment Using a Constriction Microchannel

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