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Single-cell individualized electroporation with real-time impedance monitoring using a microelectrode array chip
Microsystems & Nanoengineering ( IF 7.9 ) Pub Date : 2020-10-19 , DOI: 10.1038/s41378-020-00196-0
Zhizhong Zhang 1 , Tianyang Zheng 1 , Rong Zhu 1
Affiliation  

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.



中文翻译:

使用微电极阵列芯片进行实时阻抗监测的单细胞个体化电穿孔

在保持良好细胞活力的同时将分子精确递送到单个细胞中的能力对于治疗、诊断和药物递送的应用非常重要,因为它是朝着个性化医疗前景迈进的一步。本文报道了一种具有实时阻抗监测的单细胞个体化电穿孔方法,以使用微电极阵列芯片提高细胞穿孔效率和细胞活力。该微芯片包含多个排列成阵列的六极电极单元,用于对单个细胞进行原位电穿孔和实时阻抗监测。通过阻抗测量实时跟踪电穿孔下单个细胞的动态恢复过程,它提供了详细的瞬态细胞状态,并有助于在单个细胞水平上理解整个恢复过程。我们定义了单细胞阻抗指标来表征细胞穿孔效率和细胞活力,用于优化电穿孔。通过将所提出的电穿孔方法应用于不同的细胞系,包括单独的人类癌细胞系和正常人类细胞系,确定这些细胞的最佳刺激,从而实现增强型绿色荧光蛋白 (EGFP) 质粒进入细胞的高水平转染。结果验证了所提出的单细胞个体化电穿孔/转染方法的有效性,并证明在细胞重编程、诱导多能干细胞、过继细胞治疗、

更新日期:2020-10-19
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