Low-frequency ac electroporation shows strong frequency dependence and yields comparable transfection results to dc electroporation

Yihong Zhan, Birck Nanotechnology Center, Purdue University
Zhenning Cao, Virginia Polytechnic Institute
Ning Bao, Virginia Polytechnic Institute; Nantong University
Jianbo Li, Rutgers State University
Jun Wang, Birck Nanotechnology Center, Purdue University
Tao Geng, Birck Nanotechnology Center, Purdue University
Hao Lin, Rutgers State University
Chang Lu, Virginia Polytechnic Institute

Date of this Version



Journal of Controlled Release Volume 160, Issue 3, 28 June 2012, Pages 570–576


Conventional electroporation has been conducted by employing short direct current (dc) pulses for delivery of macromolecules such as DNA into cells. The use of alternating current (ac) field for electroporation has mostly been explored in the frequency range of 10 kHz-1 MHz. Based on Schwan equation, it was thought that with low ac frequencies (10 Hz-10 kHz), the transmembrane potential does not vary with the frequency. In this report, we utilized a flow-through electroporation technique that employed continuous 10 Hz-10 kHz ac field (based on either sine waves or square waves) for electroporation of cells with defined duration and intensity. Our results reveal that electropermeabilization becomes weaker with increased frequency in this range. In contrast, transfection efficiency with DNA reaches its maximum at medium frequencies (100-1000 Hz) in the range. We postulate that the relationship between the transfection efficiency and the ac frequency is determined by combined effects from electrophoretic movement of DNA in the ac field, dependence of the DNA/membrane interaction on the ac frequency, and variation of transfection under different electropermeabilization intensities. The fact that ac electroporation in this frequency range yields high efficiency for transfection (up to similar to 71% for Chinese hamster ovary cells) and permeabilization suggests its potential for gene delivery. (C) 2012 Elsevier B.V. All rights reserved.


Nanoscience and Nanotechnology