Search for optimum two-photon excitation method with suppressed photobleaching of fluorescent proteins
| Project/Area Number |
24560052
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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 |
Applied optics/Quantum optical engineering
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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, 教授 (80250108)
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| Co-Investigator(Renkei-kenkyūsha) |
TAKAHASHI Hiroshi 東京理科大学, 理工学部, 助教 (70572709)
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| Project Period (FY) |
2012-04-01 – 2015-03-31
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| Project Status |
Completed (Fiscal Year 2014)
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| Budget Amount *help |
¥5,330,000 (Direct Cost: ¥4,100,000、Indirect Cost: ¥1,230,000)
Fiscal Year 2014: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
Fiscal Year 2013: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
Fiscal Year 2012: ¥3,250,000 (Direct Cost: ¥2,500,000、Indirect Cost: ¥750,000)
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| Keywords | 蛍光タンパク質 / 非線形分光 / 光褪色 / 二光子励起 / バイオイメージング / 非線形光学 / 分光 |
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| Outline of Final Research Achievements |
Based on the measurement of the photobleaching action spectra of fluorescent proteins with nonlinear Fourier-transform spectroscopy, we found that the photobleaching occurs through excited-state absorption following two-photon excitation from the ground state. An unidentified dark state with a longer lifetime may be the initial state for excited-state absorption besides the triplet excited state. Then, the adaptive control method was applied to modulate the spectral amplitude and phase of excitation pulses maximizing the two-photon excited fluorescence while suppressing the photobleaching. As a result, the spectral amplitude was optimized to avoid the peak of the photobleaching spectra supporting the result of spectroscopic measurements.
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Report
(4 results)
Research Products
(19 results)
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[Presentation] Measurement of the photobleaching spectrum based on the excited-state absorption of fluorescent proteins with Fourier-transform nonlinear spectroscopy2013
Author(s)
H. Takahashi, K. Toda, K. Mochizuki, K. Isobe, F. Kannari, H. Kawano, H. Mizuno, A. Miyawaki, K. Midorikawa, and Akira Suda
Organizer
ALPS 2013
Place of Presentation
Yokohama, Japan
Related Report
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