Simultaneous maximization of cell permeabilization and viability in single-cell electroporation using an electrolyte-filled capillary

被引:11
作者
Agarwal, Aparna
Zudans, Imants
Orwar, Owe
Weber, Stephen G. [1 ]
机构
[1] Univ Pittsburgh, Dept Chem, Pittsburgh, PA 15260 USA
[2] Chalmers Univ Technol, Dept Phys Chem, SE-41296 Gothenburg, Sweden
关键词
D O I
10.1021/ac061270o
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
A549 cells were briefly exposed to Thioglo-1, which converts thiols to fluorescent adducts. The fluorescent cells were exposed to short (50-300 ms) electric field pulses (500 V across a 15 cm capillary) created at the tip of an electrolyte-filled capillary. Fluorescence microscopy revealed varying degrees of cell permeabilization depending on the conditions. Longer pulses and a shorter cell-capillary tip distance led to a greater decrease in the cell's fluorescence. Live/dead (calcein AM and propidium iodide) testing revealed that a certain fraction of cells died. Longer pulses and shorter cell-capillary tip distances were more deadly. An optimum condition exists at a cell-capillary tip distance of 3.5-4.5 mu m and a pulse duration of 120-150 ms. At these conditions, > 90% of the cells are permeabilized and 80-90% survive.
引用
收藏
页码:161 / 167
页数:7
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