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。扫描电化学显微镜（SECM）用于成像单个植物细胞的光合作用和呼吸活性，通过二维对细胞周围的局部氧浓度进行绘制。2。 SECM用于投资化学物质对单个原生质体的影响。当DCMU（将光同步的电子转移抑制剂）注入原生质体中时，原生质体的SECM图像变得阴沉并变形，因为从原生质体中降低了原生质体的氧气产生，类似地，动物细胞的呼吸活性也可以与Secm.3产生。使用双PPT微电极作为SECM的探针，以轻松检测单个原生质体附近的两个电活性物种。单个原生质体的地形图像是从Fe（CN）64-AT一个磁盘的氧化电流变化获得的。在另一个磁盘上检测局部氧浓度用于成像细胞的光合作用。4。氧气消耗量…通过SECM进行了更多的单个牛胚胎的离子。当胚胎在体外施肥后第6天到达莫拉的阶段，单个胚胎的阶段是1.40±0.27×10-14 mol s-1。 SECM测量后，将胚胎连续培养两天。发现胚泡的氧消耗率在2.50±0.46×10-14 mol S-1 <f <4.49±0.50×10-14 mol S-1.5范围内。制造了用于生物测定的微生物芯片，其性能的特征是SEC。通过映射碎片的氧气浓度，用SECM对芯片上大肠杆菌的呼吸活性进行成像。使用基于大肠杆菌的微生物芯片测量了抗生素的细菌作用。6。使用双微绝感电极进行了单个小球藻的介电性操纵。根据两个磁盘电极之间施加的交流电压频率，将有吸引力的或排斥力引起小球藻。基于介电的理论详细研究了溶液的频率和离子浓度对介电性力量对小球藻的影响。较少的

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。