2003 Fiscal Year Final Research Report Summary
EVELOPMENT OF MICROASSEY SYSTEM TO AIM AT ENVIRONMENTAL MONITORING.
| Project/Area Number |
14550775
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
生物・生体工学
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| Research Institution | Yamagata Public Corporation For the Development Of Industry, Institute Life Support Technology |
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Principal Investigator |
KASAI Shigenobu INSTITUTE FOR LIFE SUPPORT TECHNOLOGY YAMAGATA, PUBLIC CORPORATION FOR THE DEVELOPMENT OF INDUSTRY, Researcher, 生物ラジカル研究所, 研究員 (70342730)
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| Co-Investigator(Kenkyū-buntansha) |
MATSUE Tomokazu Graduate School of Environmental Studies Tohoku University, Professor, 工学部, 教授 (70173797)
TAKANO Hirohisa National Institute for Environmental, Researcher, 総合研究官 (60281698)
NODA Hiroyuki INSTITUTE FOR LIFE SUPPORT TECHNOLOGY, Researcher, 生物ラジカル研究所, 主幹研究員 (60280707)
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| Project Period (FY) |
2002 – 2003
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| Keywords | High sensitivity CCD camera / Chemiluminescence's method / SECM / Cytokine / Reactive Oxygen Species / enzyme activity |
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| Research Abstract |
1).Real-time monitoring of reactive oxygen species production during differentiation of human monocytic cell lines (THP-1)
Human monocytic leukemia cell line, THP-1, is differentiated into macrophages by phorbol myristate acetate (PMA) treatment. We report real-time monitoring of reactive oxygen species (ROSs) and cytokines production during the differentiation process. The production of ROSs by THP-1 with several hours time scale has been detected using chemiluminescence's methods.
2)Characterization of the Peroxidase Activity and hydrogen peroxide production of cell by SECM (scanning electrochemistry microscope)
The peroxidase activity in a single protoplast of alga Bryopsis plumosa is quantitatively characterized by scanning electrochemical microscopy. The generation of ferriceniummethanol (FMA^+) at the protoplast surface is directly detected by the microelectrode tip scanned close to the sample surface in seawater containing ferrocenemethanol (FMA) and hydrogen peroxide an electron mediator and an enzyme-substrate, respectively.
The macrophages consume oxygen and produce ROS which become hydrogen peroxide by a dismutation reaction. The amount of hydrogen peroxide can be measured by the current obtained when the microelectrode is brought to about 〜50μm from the macrophage surface.
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