Imaging and Manipulation of Single Living Cells with SECM

使用 SECM 对单个活细胞进行成像和操作

• 批准号: 11450323
• 负责人: MATSUE Tomokazu
• 金额: $ 9.15万
• 依托单位: Tohoku University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B).
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-11450323/
• 关键词: Imaging; SECM; Respiration; Photosynthesis; Manipulation; Bovine embryo; E-coli chip; Microelectrode; 細胞活性; 抗生物質; プロトプラスト

1. Scanning electrochemical microscopy (SECM) was used for imaging of photosynthetic and respiratory activity of a single plant cell by mapping two-dimensionally the localized oxygen concentration around the cell.2. SECM was used for investigation of the effect of chemicals on a single protoplast. When DCMU, a photosynthetic electron-transfer inhibitor, was injected into the protoplast, the SECM image of the protoplast becomes gloomy and deformed due to a decrease in the generation of oxygen from the protoplast Similarly, respiratory activity of animal cells can also be imaged with SECM.3. A dual-Pt microdisk electrode was used as a probe for SECM to simultaneously detect two electroactive species near a single protoplast. The topographic image of the single protoplast was obtained from the variation of oxidation current for Fe (CN) 64-at one disk. The detection of localized oxygen concentration at another disk was used for imaging photosynthetic activity of the cell.4. Oxygen consumpt … More ion of individual bovine embryos was noninvasively quantified by SECM.When an embryo reached the stage of a morula with a 74-μm radius on day 6 after in vitro fertilization, the oxygen consumption rate of the single morula was estimated to be 1.40±0.27×10-14 mol s-1. After the SECM measurement, the embryo was continuously cultured for another two days. The oxygen consumption rate of the blastocyst was found to be in the range of 2.50±0.46×10-14 mol s-1<F<4.49±0.50×10-14 mol s-1.5. A microbial chip for bioassay was fabricated and its performance was characterized by SECM.The respiration activity of the E.coli spot at the chip was imaged with SECM by mapping the oxygen concentration around the spot. The bactericide effects of antibiotics were measured using the E.coli-based microbial chip.6. Dielectrophoretic manipulation of a single chlorella was conducted using a dual-microdisk electrode. An attractive or repulsive force was induced to chlorella depending on the frequency of ac voltage applied between the two disk electrodes. The effect of both the frequency and ion concentration of solutions on the dielectrophoretic force on a chlorella was investigated in detail based on the theories of dielectrophoresis. Less

1.利用扫描电化学显微镜(SECM)对单个植物细胞的光合作用和呼吸活性进行了二维成像，绘制了细胞周围的局部氧浓度图。用扫描电子显微镜研究了化学物质对单个原生质体的影响。当将光合作用电子传递抑制剂DCMU注入原生质体时，由于原生质体产氧量的减少，原生质体的SECM图像变得黯淡和变形。同样，动物细胞的呼吸活动也可以用SECM.3进行成像。采用双铂微盘电极作为SECM的探针，同时检测同一原生质体附近的两个电活性物种。用单圆盘氧化铁(CN)的氧化电流变化获得了单个原生质体的形貌图像。另一个圆盘上的局部氧浓度的检测用于成像细胞的光合作用活性。耗氧量…体外受精后第6天，当胚胎发育到半径为74-μm的桑囊胚阶段时，单个桑椹胚的耗氧率为1.4 0±0.2 7×10-14 m ol S-1。SECM检测后，继续培养胚胎2天。囊胚的耗氧率为2.50±0.46×10-14moL S-1；F&lt；4.49±0.50×10-14moL S-1.5。制作了一种用于生物检测的微生物芯片，并用扫描电子显微镜(SECM)对芯片上大肠杆菌斑点的呼吸活性进行了表征，通过绘制斑点周围的氧气浓度图，利用SECM对芯片上大肠杆菌斑点的呼吸活动进行了成像。利用以大肠杆菌为基础的微生物芯片检测抗生素的杀菌效果。采用双微盘电极对单个小球藻进行了介电泳法处理。对小球藻的吸引力或排斥力取决于两个圆盘电极之间施加的交流电压的频率。根据介电理论，详细研究了溶液频率和离子浓度对小球藻介电泳力的影响。较少

项目成果

安川智之: "マイクロ電極システムを利用した単一細胞の機能探索"電気化学. 67. 264-268 (1999)

Tomoyuki Yasukawa：“使用微电极系统探索单细胞功能”电化学 67. 264-268 (1999)。

H.Shiku: "Monitoring Activity of Living Cells Entrapped in a Membrane-Sealed Microcapillary with Scanning Electrochemical Microscopy"Electrochemistry. 68. 890-892 (2000)

H.Shiku：“用扫描电化学显微镜监测膜密封微毛细管中活细胞的活性”电化学。

T.Yasukawa: "Microamperometric Measurements of Photosynthetic Activity in a Single Algal Protoplast"Biophys.J.. 76. 1129-1135 (1999)

T.Yasukawa：“单个藻类原生质体中光合活性的微电流测量”Biophys.J.. 76. 1129-1135 (1999)

T.Yasukawa: "Imaging of Photosynthetic and Respiratory Activities of a Single Algal Protoplast by Scanning Electrochemical Microscopy"Chem.Lett.. 975-976 (1999)

T.Yasukawa：“通过扫描电化学显微镜对单个藻类原生质体的光合和呼吸活动进行成像”Chem.Lett.. 975-976 (1999)

T.Yasukawa: "Characterization and Imaging of Single Cells with Scanning Electrochemical Microscopy"Electroanalysis. 12. 653-659 (2000)

T.Yasukawa：“利用扫描电化学显微镜对单细胞进行表征和成像”电分析。