Investigation of Intracellular Reactions Using Ultramicroelectrodes

使用超微电极研究细胞内反应

• 批准号: 06805075
• 负责人: MATSUE Tomokazu
• 金额: $ 1.22万
• 依托单位: TOHOKU UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1994
• 资助国家: 日本
• 起止时间: 1994 至 1995
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-06805075/
• 关键词: Ultramicroelectrode; Intracellular measurements; Protoplast; Photosynthesis; Oxygen measurements; Animal cells; Superoxide; Nitric oxide; 脂質二分子膜; ボルタンメトリー; イオンチャンネル

1. Ultramicro Pt disk electrodes with tip radii less than 5mum were fabricated via the processes ; electrochemical etching of Pt wire, sealing the Pt fine wire with a soft glass, and polishing the tip. The electrochemical characteristics of the resultant microelectrodes was investigated and found that constant potential amperometry and differential amperometry for oxygen reduction using the microelectrodes afford the oxygen concentration in the local region.2. Intracellular oxygen concentration in a protoplast from Bryopsis plumosa was determined by constant current amperometry. The intracellular oxygen reduction current increased upon light irradiation, indicating that the real-time tracing of the intracellular oxygen is possible by the present method. From the detailcd analyzes of diferential amperometric measurements using the microelectrode located near the cell, the oxygen influx or efflux was determined at a single cell level.3. The permeability of Ru (NH3) 63+ and Fe (CN) 63- through an ion channel in a planar lipid membrane was evaluated by amperometric measurements using microelectrode located near the membrane. The membrane permselectivity of the viable protoplast was also carried out by amperometry.4. Micro NO sensor and micro O_2 sensor were fabricated by immobilizing respectively Ni porphyrin and superoxide dismutase/catalase onto microelectrodes. We implanted these microsensors into rat brain and measured in vivo concentrations of NO and O_2

1。通过该过程制造的具有小于5MUM的尖端半径的超大型PT磁盘电极； PT线的电化学蚀刻，用柔软的玻璃密封PT细丝，然后抛光尖端。研究了所得微电极的电化学特性，并发现使用微电极使用微电极可提供恒定的电位安培计量法和差分放大学分析，可为局部区域提供氧气浓度。2。通过恒定电流安培法确定bry鼠原生质体中的细胞内氧浓度。在光照射后，细胞内氧还原电流增加，表明通过当前方法可以实时追踪细胞内氧的实时追踪。根据使用位于细胞附近的微电极的差异分析分析，在单个细胞水平上确定氧气涌入或外排。3。 RU（NH3）63+和Fe（Cn）63-通过平面脂质膜中的离子通道的渗透性通过使用位于膜附近的微电极测量来评估平面脂质膜中的离子通道。也通过安培法进行了可行原生质体的膜允许选择性。4。通过将Ni卟啉和超氧化物歧化酶/过氧化酶/过氧化酶固定在微电极上，制造了微无传感器和微O_2传感器。我们将这些微传感器植入大鼠脑中，并在NO和O_2的体内浓度中测量

项目成果

H.Yamada, H.Shiku, T.Matsue, I.Uchida: "Microvoltammetric Characterization of Diaphorase Monolayr at Glass Surfaces" Bioelectrochem. Bioenerg. 33. 91-93 (1994)

H.Yamada、H.Shiku、T.Matsue、I.Uchida：“玻璃表面心肌黄酶单层的微伏安特性”生物电化学。

横山英克: "NOセンサーの中枢神経領域におけるin vivo応用" 生物物理. 35. 35-36 (1995)

Hidekatsu Yokoyama：“NO 传感器在中枢神经系统中的体内应用”生物物理学 35. 35-36 (1995)。

H.Shiku, T.Takeda, H.Yamada, T.Matsue, I.Uchida: "Microfabrication and Characterization of Diaphorase-Patterned Surfaces by Scanning Electrochemical Microscopy" Anal. Chem. 67. 312-317 (1995)

H.Shiku、T.Takeda、H.Yamada、T.Matsue、I.Uchida：“通过扫描电化学显微镜对心肌酶图案表面进行微加工和表征”分析。

H.Yokoyama, N.Mori, K.Osonoe, N.Kasai, M.Hiramatsu, T.Yoshimura, T.Matsue, I.Uchida, N.Kobayashi, N.Tsuchihashi, S.Niwa: "Increase of Central Nitric Oxide During Pentylenetetrazol-Induced Seizures in Rats" Psych, Clinic, Neurosci. 49. S277-S279 (1995)

H.Yokoyama、N.Mori、K.Osonoe、N.Kasai、M.Hiramatsu、T.Yoshimura、T.Matsue、I.Uchida、N.Kobayashi、N.Tsuchihashi、S.Niwa：“中心一氧化氮的增加

H. Yokoyama: "Increase of Central Nitric Oxide During Pentylenetetrazol-Induced Seizures in Rats" Psych, Clinic, Neurosci.49. S277-S279 (1995)

H. Yokoyama：“戊四氮引起的大鼠癫痫发作期间中枢一氧化氮的增加”Psych、Clinic、Neurosci.49。