2004 Fiscal Year Final Research Report Summary
Functional Analysis of Muscle Specimen by X-ray Cryomicrodiffraction Technique
| Project/Area Number |
15500294
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Neurophysiology and muscle physiology
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| Research Institution | Japan Synchrotron Radiation Research Institute |
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Principal Investigator |
IWAMOTO Hiroyuki Japan Synchrotron Radiation Research Institute, Life & Environ.Div., Team Leader Senior Scientist, 利用研究促進部門II・生物チーム・チームリーダー, 主幹研究員 (60176568)
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| Project Period (FY) |
2003 – 2004
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| Keywords | myofibril / X-ray microdiffraction / X-ray microbeam / quick-freezing / pinhole optics / synchrotron radiation / high-flux X-ray beam / equatorial reflections |
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| Research Abstract |
The system for recording microdiffraction patterns from micrometer-sized quick-frozen hydrated biological specimens at the high-flux beamline BL40XU of SPring-8 is described. The optics consists of a pair of pinholes drilled on tantalum substratum, with a defining aperture of 2 μm in diameter. The frozen specimens are placed in an in-vacuum cryochamber mounted on a three-axis goniometer, where the specimens are stably held at a liquid-nitrogen temperature (〜-199℃). A beam size of 1.5 μm (full width at half maximum) is attained at the sample position. By using this system, diffraction patterns have been recorded from an isolated single myofibril (diameter, 〜3 μm) of an insect flight muscle in an area equivalent to a single sarcomere (length, 〜3 μm). The technique is potentially applicable to other micrometer-sized hydrated biological specimens, which are more susceptible to radiation damage than dry synthetic polymers or biopolymers. The quick-freezing of biological specimens has also been proven useful in reducing the specimen volume in the beam in conventional diffraction recordings.
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