Chen Meng-Tse, Jiang Chunqi, Vernier P Thomas, Wu Yu-Hsuan, Gundersen Martin A
Mork Family Department of Chemical Engineering and Materials Science, University of Southern California, Los Angeles, CA 90089, USA.
PMC Biophys. 2009 Nov 11;2(1):9. doi: 10.1186/1757-5036-2-9.
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-
纳秒级、兆伏每米的电脉冲可使细胞对小分子通透，导致肿瘤细胞发生程序性细胞死亡（凋亡），目前正作为一种皮肤癌治疗方法进行评估。我们使用纳米电穿孔和荧光成像技术，从三种不同的电极配置：单针、五针和平切同轴电缆，构建与向人前列腺癌细胞系PC3单层递送15纳秒、10千伏脉冲相关的二维电场图。正常情况下不透膜的荧光染料YO-PRO-1的流入作为膜通透化的敏感指标。脉冲暴露后的荧光发射水平与施加的电场强度成正比。在垂直于中心轴且距中心电极尖端15 - 20微米的平面上比较空间电场分布。测量结果与本研究评估的三种电极排列模型非常吻合。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中的电极设计提供了具有实际操作意义的校准，并允许可视化不同细胞类型对纳米电脉冲刺激的相对敏感性。物理和工程分类代码：87.85.M -
