2005 Fiscal Year Final Research Report Summary
Development of the multi-point fluorescence correlation spectroscope and analysis of molecular interaction in living cell
| Project/Area Number |
15370062
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Biophysics
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| Research Institution | Hokkaido University |
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Principal Investigator |
KINJO Masataka HokkaidcUniv. RIES, Asso. Prof., 電子科学研究所, 助教授 (70177971)
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| Co-Investigator(Kenkyū-buntansha) |
TAMURA Mamoru Hokkaido Univ. RIES, Prof, 電子科学研究所, 教授 (80089888)
NISHIMURA Goro Hokkaido Univ. RIES, Assistant, 電子科学研究所, 助手 (30218193)
MIYAZAKI Tadaaki Hokkaido Univ., Research Center for Zoonosis Control, Prof, 人獣共通感染症リサーチセンター, 教授 (60272431)
NOMURA Yasutomo Yamagata Univ. Grad. School of Sci. and Eng., Asso. Prof., 医学部, 助教授 (80237883)
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| Project Period (FY) |
2003 – 2005
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| Keywords | fluorescence correlation spectroscopy / fluorescence measurement / transcriptional factor / glucocoticoide receptor / cytosol / nuclar protein / microenvironment / laser scanning microscope |
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| Research Abstract |
Different distribution of many materials, pH, ion, or oxygen concentration can be detected in living cell according to the different position in the cell such as near or away from plasma membrane, nucleus, endsome or mitochondrion. Microenvironment of the cell is also changed according to physiological condition of the cell. The purpose of the project was construction of a multi-point measurement system of Fluorescence Correlation Spectroscopy that could detect number concentration of fluorescent molecule with the single molecule sensitivity and basic research for the detection of dynamic molecular interaction in a living cell.
At the first step of the project, we attempted to construct proto type system that could detect cross correlation function using two different fluorescence intensities. Then, detection area was increased to four positions. Finally, we reduced the duration time least 10m sec between moving position to position.
Glucocorticoid receptor, that is one of transcriptional regulator protein and traslocating protein from cytosol to nucleus after stimulated by TPA, was selected in the project. We detected the increase number of GR-GFP in nucleus and the decrease that in cytosol by TPA stimulation. The diffusion time and the fluorescence intensity per molecule of GR were analyzed using our system (MP-FCS). Furthermore, we carried out the measurement of spacial correlation analysis between two detection areas because the system could detect fluorescence emission from two positions simultaneously. When the molecules move same direction, we will be able to detect increase of correlation amplitude in image correlation function. By using this system, the dynamic movement of biomolecule will be analyzed in living cell.
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Research Products
(45 results)
