Chang Lingqian, Black Stephen, Chitrakar Chandani, Nouri Mehdi
School of Biological Science and Medical Engineering, Beihang University, Beijing, China.
Institute of Nanotechnology for Single Cell Analysis (INSCA), Beijing Advanced Innovation Center for Biomedical Engineering, Beihang University, Beijing, China.
Methods Mol Biol. 2020;2050:29-41. doi: 10.1007/978-1-4939-9740-4_4.
Electroporation has been one of the most commonly used physical methods for gene/drug delivery. Compared to other nonviral counterparts, electroporation enables optimization of delivery efficiency by tuning the electric field applied on cells. Commercial electroporation, however, results in stochastic transfection and significant cellular damage mostly due to its "bulk" environment. In this chapter, we introduce nanoelectroporation (NEP) which has demonstrated living cell transfection in a highly controllable manner. In NEP, the electric field can be precisely focused on a single cell positioned on nanochannels. Safe single-cell electroporation as well as "electrophoretic" molecular delivery can be achieved on the same device. This system achieves significantly higher transfection efficiency and cellular viability than commercial systems. This device is unique in that it can efficiently deliver genetic molecules (e.g., DNAs, RNAs) that exceed 10 kbp in size. The NEP device based on a 3D nanochannel array prototype was fabricated using cleanroom techniques. For achieving precise cell to nanochannel pairing, three on-chip high-throughput manipulation technologies were developed, that is, magnetic tweezers (MT), dielectrophoresis (DEP), and thin-film microfluidics.
电穿孔一直是基因/药物递送中最常用的物理方法之一。与其他非病毒方法相比，电穿孔可通过调节施加在细胞上的电场来优化递送效率。然而，商业电穿孔大多因其“整体”环境而导致随机转染和显著的细胞损伤。在本章中，我们介绍了纳米电穿孔（NEP），它已证明能够以高度可控的方式对活细胞进行转染。在纳米电穿孔中，电场可以精确地聚焦在位于纳米通道上的单个细胞上。在同一设备上可以实现安全的单细胞电穿孔以及“电泳”分子递送。该系统实现的转染效率和细胞活力显著高于商业系统。该设备的独特之处在于它能够有效地递送大小超过10kbp的遗传分子（例如DNA、RNA）。基于3D纳米通道阵列原型的纳米电穿孔设备是使用洁净室技术制造的。为了实现细胞与纳米通道的精确配对，开发了三种芯片上的高通量操作技术，即磁镊（MT）、介电电泳（DEP）和薄膜微流体技术。
