Sadik Mohamed M, Li Jianbo, Shan Jerry W, Shreiber David I, Lin Hao
Department of Mechanical and Aerospace Engineering, Rutgers, The State University of New Jersey, 98 Brett Road, Piscataway, NJ 08854, USA.
Biochim Biophys Acta. 2013 Apr;1828(4):1322-8. doi: 10.1016/j.bbamem.2013.01.002. Epub 2013 Jan 10.
The transport mechanisms in electroporation-mediated molecular delivery are experimentally investigated and quantified. In particular, the uptake of propidium iodide (PI) into single 3T3 fibroblasts is investigated with time- and space-resolved fluorescence microscopy, and as a function of extracellular buffer conductivity. During the pulse, both the peak and the total integrated fluorescence intensity exhibit an inverse correlation with extracellular conductivity. This behavior can be explained by an electrokinetic phenomenon known as Field-Amplified Sample Stacking (FASS). Furthermore, the respective contributions from electrophoresis and diffusion have been quantified; the former is shown to be consistently higher than the latter for the experimental conditions considered. The results are compared with a compact model to predict electrophoresis-mediated transport, and good agreement is found between the two. The combination of the experimental and modeling efforts provides an effective means for the quantitative diagnosis of electroporation.
对电穿孔介导的分子递送中的传输机制进行了实验研究和量化。具体而言，利用时间和空间分辨荧光显微镜研究了碘化丙啶（PI）进入单个3T3成纤维细胞的摄取情况，并将其作为细胞外缓冲液电导率的函数。在脉冲期间，峰值荧光强度和总积分荧光强度均与细胞外电导率呈负相关。这种行为可以用电动力学现象“场增强样品堆积”（FASS）来解释。此外，还对电泳和扩散的各自贡献进行了量化；在所考虑的实验条件下，前者始终高于后者。将结果与一个紧凑模型进行比较以预测电泳介导的传输，发现两者之间具有良好的一致性。实验和建模工作的结合为电穿孔的定量诊断提供了一种有效手段。
