Two-dimensional nanosecond electric field mapping based on cell electropermeabilization.

Meng-Tse Chen, Chunqi Jiang, P Thomas Vernier, Yu-Hsuan Wu, Martin A Gundersen
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引用次数: 28

Abstract

Nanosecond, megavolt-per-meter electric pulses cause permeabilization of cells to small molecules, programmed cell death (apoptosis) in tumor cells, and are under evaluation as a treatment for skin cancer. We use nanoelectroporation and fluorescence imaging to construct two-dimensional maps of the electric field associated with delivery of 15 ns, 10 kV pulses to monolayers of the human prostate cancer cell line PC3 from three different electrode configurations: single-needle, five-needle, and flat-cut coaxial cable. Influx of the normally impermeant fluorescent dye YO-PRO-1 serves as a sensitive indicator of membrane permeabilization. The level of fluorescence emission after pulse exposure is proportional to the applied electric field strength. Spatial electric field distributions were compared in a plane normal to the center axis and 15-20 mum from the tip of the center electrode. Measurement results agree well with models for the three electrode arrangements evaluated in this study. This live-cell method for measuring a nanosecond pulsed electric field distribution provides an operationally meaningful calibration of electrode designs for biological applications and permits visualization of the relative sensitivities of different cell types to nanoelectropulse stimulation. PACS Codes: 87.85.M-

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基于细胞电渗透的二维纳秒电场映射。
毫微秒,每米兆伏的电脉冲引起细胞对小分子的渗透,肿瘤细胞的程序性细胞死亡(凋亡),并且正在评估作为皮肤癌的治疗方法。我们利用纳米电穿孔和荧光成像技术,构建了通过三种不同的电极配置(单针、五针和平切同轴电缆)将15 ns、10 kV脉冲传递到人前列腺癌细胞PC3单层细胞时的二维电场图。通常不需要的荧光染料YO-PRO-1的内流可作为膜透性的敏感指标。脉冲照射后的荧光发射水平与外加电场强度成正比。在与中心轴垂直的平面和距离中心电极尖端15 ~ 20 μ m的平面上比较了空间电场分布。测量结果与本研究中评估的三种电极排列模型一致。这种用于测量纳秒脉冲电场分布的活细胞方法为生物应用的电极设计提供了一种有意义的校准方法,并允许可视化不同细胞类型对纳米电脉冲刺激的相对灵敏度。PACS代码:87.85.M-
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