State Research Institute, Center for Physical Sciences and Technology, Saulėtekio al. 3, Vilnius, Lithuania.
Sci Rep. 2020 Nov 5;10(1):19094. doi: 10.1038/s41598-020-76086-2.
Pulsed electric field effects induced in a membrane, as well as intracellular structures, depend on cell type, field and media parameters. To achieve desired outcomes, membranes should be permeabilized in a controlled manner, and thus efficiency of electroporation should be investigated in advance. Here, we present a framework for using mediated amperometry as a prospective method for the investigation of electroporation and its effects on cellular machinery. Whole-cell sensors with single mediator systems comprised of hydrophilic or lipophilic mediators were successfully employed to investigate membrane permeability as well as cellular responses. Exposure of yeast cells to single electric field pulse (τ = 300 µs, E = 16 kV/cm) resulted in up to tenfold increase of current strength mediated with hydrophilic mediators. Exposure to PEF resulted in decrease of menadione mediated current strength (from 138 ± 15 to 32 ± 15 nA), which could be completely compensated by supplementing electrolyte with NADH.
电场对细胞膜及细胞内结构的影响取决于细胞类型、场强和介质参数。为了达到预期的效果，细胞膜应该以可控的方式被穿孔，因此需要预先研究电穿孔的效率。在这里，我们提出了一个使用介体安培法作为研究电穿孔及其对细胞机制影响的前瞻性方法的框架。使用由亲水性或疏水性介体组成的单介体系统的全细胞传感器，成功地用于研究细胞膜通透性以及细胞反应。将酵母细胞暴露于单个电场脉冲（τ=300µs，E=16kV/cm）中，会导致亲水性介体介导的电流强度增加十倍。PEF 的暴露会导致 menadione 介导的电流强度降低（从 138±15 到 32±15nA），这可以通过用 NADH 补充电解质来完全补偿。
