1998 Fiscal Year Final Research Report Summary
Real-time in situ two-photon imaging of intercellular signaling in sinoatrial node
| Project/Area Number |
09557022
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 展開研究 |
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| Research Field |
Experimental pathology
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| Research Institution | Kyoto Prefectural University of Medicine |
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Principal Investigator |
TAKAMATSU Tetsuro Kyoto Prefectural University of Medicine, Pathology, Professor, 医学部, 教授 (40154900)
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| Project Period (FY) |
1997 – 1998
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| Keywords | Sinoatrial node / Fluo-3 / Laser scanning microscopy / Calcium ion / Digital image / Two-photon excitation / Multi-pinhole scanner |
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| Research Abstract |
The confocal microscopy has become a dependable tool for obtaining not only topological information, but also densitometric information. However, there remain many problems with the confocal microscope such as slow scanning speed, mismatched refractive indices, cell injury by the laser beam, bleaching and the penetrability of the fluorescent probe. To improve these problems, this project investigated in situ real-time imaging of intracellular calcium ion concentration ([Ca^<2+>]_i) in the cardiac ventricle or sinoatrial node by using two-photon microscopy.
We have developed a new laser scanning microscope system equipped with a microlens-arrayed multi-pinhole scanning disc, water-immersion objective lenses and a mode-locked Ti : sapphire pulse laser, which provide a scanning beam. The new system allows in situ quantitative imaging of non- homogenous changes of the three-dimensional [[Ca^<2+>]_i in a living whole heart, such as calcium wave at a full video rate. Calcium waves propagating from cell to cell were frequently interrupted by calcium transients from spontaneous sinus rhythm. But serial images obtained have low signal-to-noise and cell injury with laser light still remained unsolved.
The evolution of the confocal microscope with high spatial and temporal resolution renders the investigation of biological phenomena even in the living organ.
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