| Project/Area Number |
17206004
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| Research Category |
Grant-in-Aid for Scientific Research (A)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Thin film/Surface and interfacial physical properties
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| Research Institution | Tohoku University |
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Principal Investigator |
NIWANO Michio Tohoku University, Tohoku University, Research Institute of Electrical Communication, Professor (20134075)
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| Co-Investigator(Kenkyū-buntansha) |
KIMURA Yasuo Tohoku University, Research Institute of Electrical Communication, Assistant professor (40312673)
HIRANO Ayumi Tohoku University, Research Institute of Electrical Communication, Assistant professor (80339241)
MIYAZAKI Hitoshi University of Tsukuba, Graduate School of Life and Environmental Sciences, Professor (40183636)
ISODA Hiroko University of Tsukuba, Graduate School of Life and Environmental Sciences, Professor (00375429)
MIZOGUCHI Tsuyoshi University of Tsukuba, Graduate School of Life and Environmental Sciences, Lecturer (70281623)
石井 久夫 東北大学, 電気通信研究所, 助教授 (60232237)
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| Project Period (FY) |
2005 – 2007
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| Project Status |
Completed (Fiscal Year 2007)
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| Budget Amount *help |
¥50,050,000 (Direct Cost: ¥38,500,000、Indirect Cost: ¥11,550,000)
Fiscal Year 2007: ¥10,010,000 (Direct Cost: ¥7,700,000、Indirect Cost: ¥2,310,000)
Fiscal Year 2006: ¥14,300,000 (Direct Cost: ¥11,000,000、Indirect Cost: ¥3,300,000)
Fiscal Year 2005: ¥25,740,000 (Direct Cost: ¥19,800,000、Indirect Cost: ¥5,940,000)
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| Keywords | Cell chip / Infrared spectroscopy / Surface / Biochip / Semiconductor / 細胞 / DNA / ハイブリダイゼーション |
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| Research Abstract |
In this research, we have developed m situ real-time monitoring method for cell dynamics on the basis of infrared spectroscopy with the multiple internal reflections (MIR-IRAS). Our research projects were focused on (1) development of IRAS method for in situ monitoring of cell growth, (2) development of non-label IRAS detection of cell death and its application to cytotoxic assay. In addition, we also examined non-label detection of biomolecular interactions, such as (3) antigen-antibody interactions and (4) DNA hybridization both in solution phase and at the silicon surfaces. For the development of the real-time monitoring of cell dynamics, we started with construction of cell culture condition in a FTIR sample room. The temperature and humidity in a cell chamber placed in the FTIR sample room was controlled to be 37 °C and >80%, respectively. Since the sample room needs to be purged with air which was free from water and carbon dioxide vapor, a humidified gas mixture containing CO_2 was introduced to the chamber through silicone tubes. By using this apparatus, we have confirmed the cell viability and proliferation of HL-60 cells after 24-h incubation in the chambers. Then the cytotoxic effect by Tween20 was in-situ monitored with MIR-IRAS. It was demonstrated that the cell death can be monitored in-situ by analyzing the amide II peak intensity. In addition, we succeeded in real-time monitoring of the dynamic processes upon apoptosis induction by using MIR-IRAS. We also examined label-free detection of antigen-antibody interactions and DNA hybridization and demonstrated that MIR-IRAS is particularly useful for discriminating specific and non-specific adsorption by careful analysis of spectral features. Thus our method can be applied to in-situ monitoring of various cellular events as well as individual biomolecular interactions as non-label detection method for high-performance cell-chips and bio-chips.
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