Development of Electrical Manipulation of Single Cells Using Microelectrodes

使用微电极对单细胞进行电操作的进展

• 批准号: 08555198
• 负责人: MATSUE Tomokazu
• 金额: $ 4.86万
• 依托单位: Tohoku University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (A)
• 财政年份: 1996
• 资助国家: 日本
• 起止时间: 1996 至 1997
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08555198/
• 关键词: Microelectrode; Dielectrophoresis; Cell Manipulation; Single cell; Photosynthesis; Microelectroporation; Patterning; Membrane permeability; 電子移動; 酸化還元; 電子伝達鎖; エレクトロポレーション; 細胞配列; レドックス反応; 誘電泳動

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

1.在由微带电极组装而成的模板微阵列电极上，利用负介电泳力在玻璃基底上形成具有活细胞的微图案。将模板电极置于基底上方，并将细胞悬浮液吸入基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，10 kHz）时，诱导的介电泳力将细胞从电极上排斥并推压在玻璃基底上，形成反映电极形状的细胞图案。2.制备了用于基因或药物注射到单细胞中的微电穿孔毛细管，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。释放的Fe（CN）^3-_的量可以通过调节脉冲的幅度和周期来控制。我们将穿孔毛细管应用于 ...更多信息 将Fe（CN）_6 ~+<3->注入单个原生质体。电脉冲的施加引起了膜的可逆破裂，在膜上形成小孔，同时通过电迁移将Fe（CN）_6^<3->从毛细管中排斥出去。3.用电流法研究了几种氧化还原物种通过单个藻原生质体细胞膜的渗透。通过数值模拟定量分析氧化还原电流随微电极-膜距离的变化，确定了原生质体膜的渗透系数（Pm）。4.采用微电流法定量研究了对苯醌（BQ）对单个原生质体光合电子传递的影响。通过对BQ和对苯二酚（QH_2）定位浓度的分析，确定了单个原生质体在光照下对BQ的消耗和对苯二酚（QH_2）的产生。少

项目成果

H. Shiku: "Detection of Microspotted Carcioembryonic Antigen on a Glass Substrate by Scanning Electrochemical Microscopy." Anal. Chem.68. 1276-1278 (1996)

H. Shiku：“通过扫描电化学显微镜检测玻璃基底上的微斑点癌胚抗原。”

T.Matsue, N.Matsumoto, I.Uchida: "Rapid Micropatterning of Living Cells by Repulsive Dielectrophoretic Force" Electrochim Acta. 42. 3251-3256 (1997)

T.Matsue、N.Matsumoto、I.Uchida：“通过排斥介电泳力对活细胞进行快速微图案化”电化学学报。

末永智一: "走査型電気化学顕(SECM)による局所センシング" 化学センサ. 13. 8-13 (1997)

Tomokazu Suenaga：“使用扫描电化学显微镜 (SECM) 进行局部传感”化学传感器。13. 8-13 (1997)

T.Yasykawa: "Permeation of Redox Species Through a Cell Membrane of a Single,Living Algal Protoplast Studied by Microamperometry" Biochim.Biophys.Acta.(in press).

T.Yasykawa：“通过微电流分析法研究氧化还原物质通过单个活藻类原生质体细胞膜的渗透”Biochim.Biophys.Acta.（出版中）。

H.Shiku: "Dual lmmunoassay of Human Chorionic Gonadotropin and Human Placental Lactogen at a Microfabricated Substrate by Scanning Electrochemical Microscopy" J.Electroanal.Chem.438. 187-190 (1997)

H.Shiku：“通过扫描电化学显微镜在微型基底上对人绒毛膜促性腺激素和人胎盘催乳素进行双重免疫测定”J.Electroanal.Chem.438。