1996 Fiscal Year Final Research Report Summary
Manufacture of Scanning Force Microscope for Biological Research
| Project/Area Number |
06558099
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| Research Category |
Grant-in-Aid for Scientific Research (A)
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| Allocation Type | Single-year Grants |
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| Section | 試験 |
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| Research Field |
Biophysics
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| Research Institution | Kanazawa University |
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Principal Investigator |
ANDO Toshio Kanazawa University, Science, Professor, 理学部, 教授 (50184320)
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| Co-Investigator(Kenkyū-buntansha) |
TAKENOBU Takayoshi Olympus, 2nd R/D,Researcher, 第2開発部, 研究員
HAYASHI Yoshiaki Olympus, 2nd R/D,Chief, 第2開発部, 係長
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| Project Period (FY) |
1994 – 1996
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| Keywords | Atomic Force Microscope / AFM / High-speed Scanner / High-speed Scanning / Protein / Bi-molecular Interaction / Myosin Heads |
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| Research Abstract |
(a) We manufactured an atomic force microscope that was able to be scanned faster and less susceptible to drift than ordinary AFM apparatuses. This AFM was attached to an epiluminescence fluorescence microscope. To show the high performance of our AFM apparatus we tried to obtain high resolution images of myosin heads (subfragment-1) in aqueous solution. The images revealed fine structures that was comparable to those in an atomic model of S1 derived from x-ray crystallography of S1. In the AFM images we were able to see the ATP binding site as well as the large cleft at the tip of S1. These results proved that our AFM apparatus was on the level of the greatest in the world.
(b) We performed a study preparatory to the development of a high-speed AFM in which one image would be acquired within 30msec. We designed and manufactured a high-speed scanner and attained high-speed imaging where one image was obtained within 150msec. By way of experiment we produced an AFM that employed a displacemant detection system suitable for high-speed imaging. We also examinds methods for preparing cantilevers that were suitable for a high-speed AFM.From these preparative studies we were able to secure a scaffolding for the development of the video-rate AFM.
(c) To measure bimolecular interaction force at a truly single molecular level we developed a method to capture a single molecule of protein at the apex of a cantilever tip. Using this method we succeeded in quantifying the force field exerted between a single molecule of heavy meromyosin (HMM) and actin. Using this new technology we also discovered that ATP binding to HMM heads induced vibratile structural changes in HMM heads.
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