Development of Digital Enzyme Device

数字化酶装置的开发

• 批准号: 06558117
• 负责人: MATSUE Tomokazu
• 金额: $ 4.67万
• 依托单位: TOHOKU UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Developmental Scientific Research (B)
• 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/grant/KAKENHI-PROJECT-06558117/
• 关键词: Ultramicroelectrode; Intracellular measurement; Protoplast; Photosynthesis; Oxygen measurements; Animal cells; Superoxide; Nitric oxide; ポリピロール; ポリアニリン; フラビン酵素; グルコースオキシダーゼ; ジアフォラーゼ; ペニシリナーゼ

1.Ultramicro Pt disk electrodes with tip radii less than 5 mum 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 dctailed analyzes of differential 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 implated these microsensors into rat brain and measured in vivo concentrations of NO and O_2.

1.超微铂盘电极的尖端半径小于5微米，通过制造的过程;电化学蚀刻的铂线，密封的铂细线与软玻璃，和抛光的尖端。研究了微电极的电化学特性，发现微电极用于氧还原的恒电位安培法和微分安培法可以提供局部区域的氧浓度。2.采用恒电流安培法测定了羽状真藓原生质体细胞内的氧浓度。细胞内氧还原电流在光照下增加，表明本方法可以实时跟踪细胞内氧。采用微电极差示电流法测定了Ru（NH3）63 +和Fe（CN）63-在脂膜离子通道中的渗透性。4.将Ni卟啉和超氧化物歧化酶/过氧化氢酶分别固定在微电极上，制备了微型NO传感器和微型O_2传感器。我们将这些微传感器植入大鼠脑内，并在体内测量NO和O_2的浓度。

项目成果

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。

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

M.Shibuya, T.Hasegawa, T.Matsue, I.Uchida: "Electrochemical Behavior of V-Grrove Microelectrodes Fabricated by Micromachining" J.Electroanal. Chem. 396. 535-540 (1995)

M.Shibuya、T.Hasekawa、T.Matsue、I.Uchida：“微加工制造的 V-Grrove 微电极的电化学行为”J.Electroanal。

H. Yokoyama: "Direct and Continuous Monitoring of Intahippocampal Nitric Oxide(NO)by an NO Senser in Freely Moving Rat after N-Methyl-D-aspartic Acid Injection" Denki Kagaku. 63. 1167-1170 (1995)

H. Yokoyama：“注射 N-甲基-D-天冬氨酸后，通过 NO 传感器对自由活动的大鼠海马一氧化氮 (NO) 进行直接连续监测”Denki Kagaku。

N.Kasai, T.Matsue, I.Uchida: "Multichannel Electrochemical Analyzes of Enzyme Reactions" Electroanalysis. (in press).

N.Kasai、T.Matsue、I.Uchida：“酶反应的多通道电化学分析”电分析。