Fast electrical lysis of cells for capillary electrophoresis

被引:88
作者
Han, FT
Wang, Y
Sims, CE
Bachman, M
Chang, RS
Li, GP [1 ]
Allbritton, NL
机构
[1] Univ Calif Irvine, Dept Physiol & Biophys, Irvine, CA 92697 USA
[2] Univ Calif Irvine, Integrated Nanosyst Res Facil, Irvine, CA 92697 USA
[3] Univ Calif Irvine, Dept Elect & Comp Engn, Irvine, CA 92697 USA
[4] Univ Calif Irvine, Ctr Biomed Engn, Irvine, CA 92697 USA
关键词
D O I
10.1021/ac0341970
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
In the past decade, capillary electrophoresis has demonstrated increasing utility for the quantitative analysis of single cells. New applications for the analysis of dynamic cellular properties demand sampling methods with sufficient temporal resolution to accurately measure these processes. In particular, intracellular signaling pathways involving many enzymes can be modulated on subsecond time scales. We have developed a technique to rapidly lyse an adherent mammalian cell using a single electrical pulse followed by efficient loading of the cellular contents into a capillary. Microfabricated electrodes were designed to create a maximum voltage drop across the flattened cell's plasma membrane at a minimum interelectrode voltage. The influence of the interelectrode distance, pulse duration, and pulse strength on the rate of cell lysis was determined. The ability to rapidly lyse a cell and collect and separate the cellular contents was demonstrated by loading cells with Oregon Green and two isomers of carboxyfluorescein. All three fluorophores were detected with a separation efficiency comparable to that of standards. Parallel comparison of electrical lysis to that produced by a laser-based lysis system revealed that the sampling efficiencies of the two techniques were comparable. Rapid cell lysis by an electrical pulse may increase the application of capillary electrophoresis to the study of cellular dynamics requiring fast sampling times.
引用
收藏
页码:3688 / 3696
页数:9
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