Development of Electrical Manipulation of Single Cells Using Microelectrodes

1.利用由微带电极组成的模板微阵列电极诱导的负介电泳力，在玻璃衬底上形成带有活细胞的微图案。模板电极被置于衬底之上，细胞悬浮液被吸入衬底和电极之间的空间。当在模板电极上施加交流电压（通常为7Vrms， 10kHz）时，诱导的介电泳力将细胞从电极上排斥出去，并推动玻璃基板形成反映电极形状的细胞图案。制备了用于基因或药物注射的微电孔毛细管，并对其性能进行了研究。当施加电脉冲时，溶解在毛细管内溶液中的Fe (CN) ^3-_通过电迁移释放出来。通过调节脉冲的大小和周期，可以控制Fe (CN) ^3-_的释放量。我们利用毛细管孔法将Fe (CN) _6^<3->注入单个原生质体中。电脉冲作用使膜发生可逆击穿，膜上形成小孔隙，同时电迁移使Fe (CN) _6^<3->从毛细管中排出。用安培法研究了几种氧化还原物质通过单一藻类原生质体细胞膜的渗透情况，并在细胞膜附近放置了一个铂微盘电极。通过数字模拟定量分析氧化还原电流随微电极-膜距离的变化，确定原生质体膜的渗透系数（Pm）。采用微安培法定量研究了对苯醌（BQ）对光合作用电子传递的影响。通过对BQ和对对苯二酚（QH_2）局部浓度的分析，确定了光照射下单个原生质体对BQ的消耗和QH_2的生成量。少

1.Micropatterns with living cells were formed on glass substrates using negative dielectrophoretic force induced at a template microarray electrode consisting of assembly of microband electrodes. The template electrode was placed above the substrate and cell suspension was sucked into the space between the substrate and electrode. When an ac voltage (typically, 7Vrms, 10kHz) was applied to the template electrode, the induced dielectrophoretic force repelled the cells from the electrode and pushed at the glass substrate to form the cell pattern which reflected the electrode shape.2.Microelectroporation capillary for injection of genes or drugs into single cells were fabricated and the performance was investigated. When an electric pulse was applied, Fe (CN) ^3-_ dissolved in the solution inside the capillary was released by electromigration. The amount of the released Fe (CN) ^3-_ can be controlled by adjusting the magnitude and period of the pulse. We applied the poration capillary to … More inject Fe (CN) _6^<3-> into a single protoplast. The application of a electric pulse brought about a reversible membrane breakdown to form small pores in the membrane and simultaneously repelled Fe (CN) _6^<3-> from the capillary by electromigration.3.Permeation of several redox species through a cell membrane of a single algal protoplast was investigated by amperometry with a Pt microdisk electrode located near the membrane. Permeability coefficient (Pm) of the protoplast membrane was determined by the quantitative analysis of the variation of the redox current with microelectrode-membrane distance using digital simulation.4.Miceoamperometric measurements of a single protoplast were applied for quantitative investigation of the influence of p-benzoquinone (BQ) on the photosynthetic electron-transport. From the analyzes of the localized concentrations of BQ and p-hydroquinone (QH_2) , the consumption of BQ and the generation of QH_2 by a single protoplast upon light irradiation were determined. Less