Abstract

The electroporation of cells is nowadays used for a large variety of purposes, from basic research to cancer therapy and food processing. Understanding molecular mechanisms of the main processes involved in electroporation is thus of significant interest. In the present work, we propose an experimental system to record in real time the evolution of any cell parameter which can be evaluated by fluorescence (before, during and after application of the electroporation pulses to cells in suspension). The system is based on the design of adequate electroporation electrodes, compatible with a standard spectrofluorometer cuvette housing. The electric field intensity generated when pulses are applied was carefully characterized for different geometries of the electrodes, to choose a construction ensuring the greatest homogeneity of the field in combination with the best possible illumination of the sample. As an example of the method’s application, we present here generalized polarization kinetics for a varying number of electroporation pulses applied to a cell suspension; the general polarization parameter is strongly correlated to water presence in the hydrophobic membrane core. The system may be used for many other fluorescence measurements useful for the characterization of the electroporation process.

中文翻译：

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一种实时荧光记录电穿孔引起的细胞膜变化的实验系统

摘要

细胞电穿孔现在用于多种用途，从基础研究到癌症治疗和食品加工。因此，了解电穿孔主要过程的分子机制具有重要意义。在目前的工作中，我们提出了一个实验系统来实时记录可以通过荧光评估的任何细胞参数的演变（在将电穿孔脉冲应用于悬浮细胞之前、期间和之后）。该系统基于适当的电穿孔电极的设计，与标准的荧光分光光度计比色皿外壳兼容。对不同几何形状的电极仔细表征施加脉冲时产生的电场强度，选择一种结构，确保最大的场均匀性与样品的最佳照明相结合。作为该方法应用的一个例子，我们在这里展示了应用于细胞悬浮液的不同数量的电穿孔脉冲的广义极化动力学；一般极化参数与疏水膜核心中的水存在密切相关。该系统可用于许多其他对电穿孔过程表征有用的荧光测量。一般极化参数与疏水膜核心中的水存在密切相关。该系统可用于许多其他对电穿孔过程表征有用的荧光测量。一般极化参数与疏水膜核心中的水存在密切相关。该系统可用于许多其他对电穿孔过程表征有用的荧光测量。