纳米电穿孔具有电压低、孔径小、精度高的优点，能够在高效导入外源分子的同时最大程度地保留细胞的活性和功能，是一种前景广阔的细胞转染新方法。然而目前对于纳米电穿孔的机理研究尚不完善，缺乏有效的解决方案和实验方法以提高转染过程的导入率和细胞存活率，限制了这项技术的实用化。本项目在前一个自然基金（51575090）的基础上，面向高效可控细胞转染的需求，系统性研究电场介导细胞膜孔的形成机理和外源分子导入培养细胞的动力学特性，建立纳米电穿孔过程动力学模型；改进现有纳米电穿孔的电极设计和微流控芯片的制造材料与工艺，设计并制造一种可控性强、导入率高、细胞毒性小的纳米电穿孔微流控芯片；结合理论模型和实验，探索精确控制膜孔形成与闭合、介导位置与剂量，提高导入率和细胞存活率的方法。本项目有望推动纳米电穿孔在细胞转染中的应用，为发展更为可靠的单细胞转染技术提供有益的借鉴。

Due to the advantages of low applied voltage, localized electrical field, and high precision, nanoelectroporation is capable of efficiently delivering molecules into cells while maintaining their viability and functions which is a promising technology for cell transfection. However, a lack of a mechanistic understanding of the electroporation process hinders its implementation in sampling and transfection. In this work, we investigate the mechanisms of localized electrical field induced pore evolution and closing, and the transportation dynamics of molecules, and develop a multiphysics model of localized electroporation and molecular transport to explain the delivery of molecules in live cells. Then optimize the design, fabrication processes, and materials to develop a lab-on-a-chip platform with nanostructured electrodes to improve the transfer efficiency and viability. Finally, numerical simulations combined with experiments are performed to explore new methodologies to control the pores evolution as well as the position and dosage of electroporation. This work will advance the application of nanoelectroporation in application to cell transfection and give insight into developing more reliable single cell transfection techniques.