Transfection of cells using flow-through electroporation based on constant voltage

Tao Geng, Birck Nanotechnology Center, Purdue University
Yihong Zhan, Birck Nanotechnology Center, Purdue University
Jun Wang, Birck Nanotechnology Center, Purdue University
Chang Lu, Virginia Polytechnic Institute

Date of this Version



Nature Protocols 6, 1192–1208 (2011)


Electroporation is a high-efficiency and low-toxicity physical gene transfer method. Classical electroporation protocols are limited by the small volume of cell samples processed (less than 107 cells per reaction) and low DNA uptake due to partial permeabilization of the cell membrane. Here we describe a flow-through electroporation protocol for continuous transfection of cells, using disposable devices, a syringe pump and a low-cost power supply that provides a constant voltage. We show transfection of cell samples with rates ranging from 40 mu l min(-1) to 20 ml min(-1) with high efficiency. By inducing complex migrations of cells during the flow, we also show permeabilization of the entire cell membrane and markedly increased DNA uptake. The fabrication of the devices takes 1 d and the flow-through electroporation typically takes 1-2 h.


Nanoscience and Nanotechnology