Batista Napotnik Tina, Miklavčič Damijan
University of Ljubljana, Faculty of Electrical Engineering, Tržaška 25, 1000 Ljubljana, Slovenia.
University of Ljubljana, Faculty of Electrical Engineering, Tržaška 25, 1000 Ljubljana, Slovenia.
Bioelectrochemistry. 2018 Apr;120:166-182. doi: 10.1016/j.bioelechem.2017.12.005. Epub 2017 Dec 16.
Exposing cells to an electric field leads to electroporation of the cell membrane which has already been explored and used in a number of applications in medicine and food biotechnology (e.g. electrochemotherapy, gene electrotransfer, extraction of biomolecules). The extent of electroporation depends on several conditions, including pulse parameters, types of cells and tissues, surrounding media, temperature etc. Each application requires a specific level of electroporation, so it must be explored in advance by employing methods for detecting electroporation. Electroporation detection is most often done by measuring increased transport of molecules across the membrane, into or out of the cell. We review here various methods of electroporation detection, together with their advantages and disadvantages. Electroporation detection can be carried out by using dyes (fluorophores or colour stains) or functional molecules, by measuring the efflux of biomolecules, by impedance measurements and voltage clamp techniques as well as by monitoring cell swelling. This review describes methods of detecting cell membrane electroporation in order to help researchers choose the most suitable ones for their specific experiments, considering available equipment and experimental conditions.
将细胞暴露于电场会导致细胞膜发生电穿孔,这一现象已在医学和食品生物技术的许多应用中得到探索和应用(例如电化学疗法、基因电转移、生物分子提取)。电穿孔的程度取决于多种条件,包括脉冲参数、细胞和组织类型、周围介质、温度等。每种应用都需要特定水平的电穿孔,因此必须通过采用检测电穿孔的方法预先进行探索。电穿孔检测通常是通过测量分子跨膜进入或离开细胞的运输增加来进行的。我们在此回顾各种电穿孔检测方法及其优缺点。电穿孔检测可以通过使用染料(荧光团或彩色染色剂)或功能分子、测量生物分子的外流、通过阻抗测量和电压钳技术以及监测细胞肿胀来进行。本综述描述了检测细胞膜电穿孔的方法,以帮助研究人员根据可用设备和实验条件为其特定实验选择最合适的方法。