| Project/Area Number |
60880027
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| Research Category |
Grant-in-Aid for Developmental Scientific Research
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| Allocation Type | Single-year Grants |
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| Research Field |
分子遺伝学・分子生理学
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| Research Institution | Institute of Biological Sciences, University of Tsukuba |
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Principal Investigator |
ISHIZAKA Shozo Inst. of Biological Sciences, Univ. of Tsukuba, Professor, 生物科学系, 教授 (10062499)
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| Co-Investigator(Kenkyū-buntansha) |
KOBAYASHI Katsumi National Laboratory for High Energy Physics, Photon Factory,Assitant Professor, 放射光実験施設, 助教授 (20114077)
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| Project Period (FY) |
1985 – 1986
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| Project Status |
Completed (Fiscal Year 1986)
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| Budget Amount *help |
¥8,500,000 (Direct Cost: ¥8,500,000)
Fiscal Year 1986: ¥2,500,000 (Direct Cost: ¥2,500,000)
Fiscal Year 1985: ¥6,000,000 (Direct Cost: ¥6,000,000)
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| Keywords | non-destructive cell sorting / dynamic morphometry / optical transformation / marking on specific sites interference fringes on diffraction pattern / Fourier transformation / 電算シミュレーション / フーリエ変換 / 刺激に対する応答の畳込みを解く / レーザー・フーリエ変換 / 形態情報 / 電算画像 / 散乱点集合 / 抗原抗体被覆 / マイクロスフィア標識 / 特異部位の分布 / 細胞選別 |
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| Research Abstract |
Sorting a cell with specific genes out of the culture is biologically aquired now. The amount of the product produced material by the cell is too small to detect directly. However, the conformation change and the response against stimulus are frequently affeced by the small difference of the genes. A readymade cell sorter which uses an advanced non-distructive detection by means of laser was adopted. This worked on the basis of optical transformation and Fourier transformation which give the laser scattered laser intensiy and strength of the material cell, from which the discrimination of the conformation changes and its response against stimulus has been examined for the dynamic morphometry.
One of the features of the cells is the distribution of specific sites. Human lymphocytes were used as the material to detect the spacing of its IgG sites on the cell surface. Microspheres of high refractive polystyrene were coated with antibodies and marked on the IgG sites by antigen-antibody rea
… More
ction. The diffraction pattern was obtained by irradiation of laser light on each distributed microsphere. The pattern shows a set of concentric circular fringes with the center at the optical axis. Superimposed on the patterns are sets of coaxial fringes from which the spacing of the sites can be figured out. The distance between a pair of microspheres is indicated by the density of the interference fringes and the direction of the relative position can be known from the parallel axis to the common axis of the fringes. The results coincide with the microscopic photographs of the cell.
Then, if the diffraction pattern and the orientation of the markers are known, the spacing of markers of any form can be determined from its interference fringes. First, the diffraction pattern of any shape which is difficult to analyze was simulated by the help of a computer with its accuracy and checked whether it could be distinguished with 1/4. The detection of the spacing with this accuracy makes morphometry of specific sites of chromosomes possible in a few microseconds.
On the basis of the computer simulation of the diffraction pattern of any marker, fluid orientation of the markers has been observed. The marker material was a meniscus bead. The beads were placed into a flow in a tube with rectangular cross section. Laser beam was irradiated perpendicularly to the flow. The scattered light on the flowing beads was detected through the optical filter. The data were integrated by the computer in correlation to the orientations of the marker and the rectangular cross section.
As a result, the optical transformation of cell organella could be used for morphometry. Since the Fourier component of the conformation change is the product of the Fourier component of the given stimulus and the Fourier component of the response function convoluting with the stimulus, a new age of the non-distructive sorting will be developed by dynamic morphometry. Less
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