| Project/Area Number |
16K04983
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Multi-year Fund |
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| Section | 一般 |
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| Research Field |
Optical engineering, Photon science
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| Research Institution | Tokyo University of Science |
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Principal Investigator |
SUDA AKIRA 東京理科大学, 理工学部物理学科, 教授 (80250108)
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| Project Period (FY) |
2016-04-01 – 2019-03-31
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| Project Status |
Completed (Fiscal Year 2018)
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| Budget Amount *help |
¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000)
Fiscal Year 2018: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000)
Fiscal Year 2017: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000)
Fiscal Year 2016: ¥2,860,000 (Direct Cost: ¥2,200,000、Indirect Cost: ¥660,000)
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| Keywords | 深部観察 / 多光子過程 / 蛍光顕微鏡 / 超解像イメージング / 応用光学・量子光工学 / 非線形光学 / バイオイメージング |
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| Outline of Final Research Achievements |
In order to realize high-resolution fluorescence imaging in the deep part of living tissue, we have developed a two-photon excitation technique for intentionally bleaching fluorescent proteins, as well as wide-field two-photon imaging with spatio-temporal focusing. As a result of measuring the photobleaching rate of eGFP, the amount of change with or without the Excited-State Absorption (ESA)-induced light was small, which suggested that it was difficult to control the photobleaching to the extent necessary for two-photon excited super-resolution imaging. In order to clarify this cause, we constructed a total internal reflection fluorescence microscope to observe the fluorescent proteins at single molecule level. It was found that charge-transfer ESA causes reversible blinking, and irreversible photobleaching is a secondary phenomenon which is related to the blinking.
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| Academic Significance and Societal Importance of the Research Achievements |
蛍光タンパク質の褪色現象はその原因が明らかでないなか、超解像観察などにおいて積極的に利用されている。本研究では、深部観察に有効な二光子蛍光観察において褪色を利用することを試みたが、超解像観察に必要な程度まで褪色を制御することは困難であった。励起状態吸収に続く電荷移動が可逆的な光明滅を引き起こしていること、不可逆的な光褪色はそれに連動して生じる副次的な現象であることが原因と考えられ、これは褪色現象のさらなる解明と応用の展開に繋がるものである。
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