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FUNDING PROGRAM FOR NEXT GENERATION WORLD-LEADING RESEARCHERS Development of a novel single-molecule imaging technique using fluorescent diamond nanoparticles and its Project Title: application to biomolecule observation Name: Yoshie HARADA Institution: Kyoto University 1. Background of research In order to understand biological activity, it is very important to clarify functional process of each protein molecule in a complicated cellular space. The conventional tool to observe the dynamics and change in the structural conformation for proteins is single molecule fluorescence microscopy, where a fluorescent "tag" (it is a special and very small molecule to emit light) is labeled on the target protein and is traced in time and space. However, the fluorescent tags currently available have a couple of fundamental problems; (1) unstable photophysical property (prevent observation for a long time), (2) difficult discrimination of the tag’s light from pre-existing light sources and (3) impossible observation of minute rotating motion of protein. 2. Research objectives We propose a new methodology by use of very small molecular sized diamond particles as a new fluorescence tag. Responsible for the fluorescence of the particles is a single nitrogen atom paired with a vacancy of carbon atom in diamond. The establishment of this new fluorescence microscope able to observe novel biological activity is our primary goal of this project. 3. Research characteristics (incl. originality and creativity) This diamond fluorescence tag has properties superior to the widely used tags now a day, and is anticipated to clear up all the problems mentioned above. Consequently, we are confident to measure new dynamics and structural change of individual protein that have been unable to observe so far. 4. Anticipated effects and future applications of research The accumulation of new knowledge about functional process of proteins provides not only profound understanding of biological activity but also important information regarding medical treatment and drug discovery for diseases, the parts of which are caused by malfunctioning of activity of certain proteins. Clarification of Function of Protein Understand Biological Activity Observation by Single Molecule Fluorescence Microscope 【Problems in Observation】 • unstable emission phenomena • segregation from inherit fluorescence • difficult observation of rotating motion • highly toxic 【Fluorescent Diamond】 • stable emission phenomena • eliminate inherit fluorescence • quantitative measurement of rotation • low toxicity New Instrumentation of Fluorescence Imaging Single Molecule Fluorescence Methodology Magnetic Resonance Methodology Combination of Single Molecule Observation and Structural Biology • Understanding disease at molecular scale • Application to drug discovery and therapy
