Abstract Electroporation technique has recently emerged as a tool for delivery of foreign molecules into cells. However, the electroporation has many critical hurdles to overcome in cell viability, delivery efficiency, and productivity. To overcome the hurdles with a single platform, we devised a polyimide (PI) film-based on- chip electroporation system that shields the cells from the electrodes with four sheath flows, enabling a 3D flow focusing. This on-chip electroporation with a double forced-flow (OE-DFF) configuration enhances the cell viability to such an extent that even with a long spiral channel for high molecular delivery efficiency, which is detrimental to the cell viability due to longer exposure to the electric field, the cell viability is still increased substantially. The advantages provided by the OE-DFF system is demonstrated with a fluorescent probe molecule (FITC-BSA) and pPtCrCFP plasmid delivered into Chlamydomonas reinhardtii, one of the challenging cell lines to transform. The continuous nature of the flow system assures high throughput. This novel approach in microfluidic science is expected to greatly contribute to algal research as an efficient electroporation tool as well as to broad applications.

中文翻译：

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具有鞘流设计的聚酰亚胺薄膜片上电穿孔系统，可将分子有效输送到微藻中

摘要 电穿孔技术最近已成为一种将外来分子输送到细胞中的工具。然而，电穿孔在细胞活力、递送效率和生产力方面有许多关键障碍需要克服。为了克服单一平台的障碍，我们设计了一种基于聚酰亚胺 (PI) 膜的片上电穿孔系统，该系统通过四个鞘流保护细胞免受电极影响，从而实现 3D 流聚焦。这种具有双强制流 (OE-DFF) 配置的片上电穿孔可将细胞活力提高到这样的程度，即使使用长螺旋通道来实现高分子递送效率，由于长时间暴露于环境中，这对细胞活力不利。在电场作用下，细胞活力仍然大幅增加。OE-DFF 系统提供的优势通过将荧光探针分子 (FITC-BSA) 和 pPtCrCFP 质粒传递到莱茵衣藻中得到证明，莱茵衣藻是一种具有挑战性的转化细胞系。流动系统的连续特性确保了高吞吐量。预计这种微流体科学中的新方法将极大地促进藻类研究，作为一种有效的电穿孔工具以及广泛的应用。