2017 Fiscal Year Annual Research Report
Anisotropic magnetic field effect imaging microspectroscopy: a technique to search cells for magnetic compass ability
| Project/Area Number |
17H03005
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| Research Institution | The University of Tokyo |
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Principal Investigator |
ウッドワード ジョナサン 東京大学, 大学院総合文化研究科, 准教授 (80526054)
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| Project Period (FY) |
2017-04-01 – 2020-03-31
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| Keywords | 磁気受容 / ラジカル反応 / 磁場効果 / 磁場効果 / スピン化学 |
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| Outline of Annual Research Achievements |
During the first year of the project, a number of key milestones were achieved in terms of experimental work, theoretical calculations and publications. The TIRF microscope has been constructed and has undergone many modifications to optimize all aspects of the signal path, critical to observing magnetic field effects on single molecules.
The projected field electromagnet was built by GMW Associates in the United States and installed into the TIRF microscope. Testing has confirmed that no artifactual signals were introduced in the optical datection system.
Preliminary methods for constraining flavin adenine dinucleotide (FAD) molecules for single molecule and anisotropic magnetic field effect experiments have been developed. These are based on encapsulation within agarose and polyacrylamide gels.
Theoretical calculations tp determine the necessary conditions for observating magnetic field effects (MFEs) on single FAD radical pairs were submitted to the journal Molecular Physics in collaboration scientists at Saitama University, Japan, Novosibirsk State University, Russia and the International Tomography Center, Russia.
Posters were presented at three conferences in 2017 and some of the work was presented orally at two conferences in 2017.
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| Current Status of Research Progress |
Current Status of Research Progress
2: Research has progressed on the whole more than it was originally planned.
Reason
The microscope is fully operational. Currently simple experiments are underway to measure magnetic field effects (MFEs) on FAD photochemistry using the new magnet system.
In order to observe single molecule MFEs on FAD, the currently developed methods still allow too much molecular movement, so alternative methods need to be developed. In addition, our recent calculations indicate that greater laser power is necessary to drive the single molecule cycle to a state where MFEs become visible.
Cell culturing has been established and preliminary experiments on HeLa cell FAD autofluorescence have begun. Transfection of the cells to allow over-expression of cryptochrome proteins has not yet been undertaken.
Realistic kinetic simulations for FAD single molecules are completed.
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| Strategy for Future Research Activity |
Record FAD MFEs using the new microscope and projected field magnet and publish technical paper in collaboration with GMW Associates.
Optimise existing and develop new methods for FAD molecular confinement. Once achieved, measure anisotropic MFEs in this system.
Purchase and install new CW pump laser system to achieve the required laser intensities at the sample.
Construct dual path camera / single photon counting detection system to allow single molecule MFE measurements to be undertaken. Make measurements using the confined FAD samples.
Undertake measurements of MFEs on FAD autofluorescence in HeLa cells.
Develop realistic Liouville space spin dynamic simulations for simulating MFEs on single FAD molecules.
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