| Budget Amount *help |
¥5,590,000 (Direct Cost: ¥4,300,000、Indirect Cost: ¥1,290,000)
Fiscal Year 2014: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2013: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2012: ¥2,730,000 (Direct Cost: ¥2,100,000、Indirect Cost: ¥630,000)
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| Outline of Final Research Achievements |
Fluorescence imaging by two-photon excitation has attracted attention as a new imaging technique. However, fluorescence mechanism of two-photon excited fluorescent proteins is completely unknown. We measured two-photon absorption spectra of enhanced GFP (eGFP), which is the most important marker, and its model chromophore, HBDI, in various solvents by a multiplex two-photon absorption spectroscopy to clarify the fluorescence mechanism. The results indicate that the existence of a "hidden" excited state in the vicinity of the lowest excited singlet state of eGFP. We conclude that this is the origin of the discrepancy between the one-photon and two-photon excitation spectra of eGFP, which is well known in the field of biology.
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