1987 Fiscal Year Final Research Report Summary
Image analysis of weakly birefringnet cytoskeletons recorded by high resolution DIC-Pol microscope and a sensitive time-lapsc VTR combined with image analyzer.
| Project/Area Number |
60480020
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
動物発生・生理学
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| Research Institution | School of Science, Nagoya University |
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Principal Investigator |
SATO Hidemi Sugashima Marine Biological Loboratory, Nagoya University, 理学部, 教授 (40109260)
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| Co-Investigator(Kenkyū-buntansha) |
SATO Yukiko Biological Laboratory, Shoin Women's University, 一般教育, 助教授 (80123636)
KATO Toyoki Sugashima Marine Biological Laboratory, Nagoya University, 理学部, 助手 (40115548)
KURODA Hideyo Sugashima Marine Biological Loboratory, Nagoya University, 理学部, 助教授 (50064845)
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| Project Period (FY) |
1985 – 1987
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| Keywords | Weak Birefringence / Cytoskeleton / Mitotic Spindle / Microtubules |
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| Research Abstract |
In general, Cell motility and any vectorial movement of cellular organelles in eukaryotes were supposed by regulated by the cytoskeletons.
Due to the nature of molecular alignment or the arrangement of fine structures, these cytoskeletons commomly gained an optical anisotropy and yielded birefringence (BR). However, their BR were weak and fructuated reflecting the labile molecular association or polymerized states in living cells.
For the purpose to obtain reliable reliable and analyzable data using living material, we devised an equipment which allowed us to measure BR and also record images on VTE. This equipment consisted of: A Nikon model HPD Nomarski-Polarization microscope equipped with rectified condensor and a Brace-Koehler type, rotating mica compensator (Nippon Kogaku K. K., Tokyo); a Panasonic model WV 1900 camera designed specifically record subjects in low light intensity (nicknamed as "moonlight" TV camera) and model NV 8050 time-lapse VTR (Matsushita Denki K. K., Yokohama)
… More
;"Image "Image analyzer with digital analog converter (Nihon avionics Co., Osaka); even and strong illuminations at 546 to 550 nm filteredfrom halogen 100 W lamp or osram HBO 100 W mercury arc. This combination of instruments worked well and We made following experiments.
1) We recorded the process of assmbly and disassmbly of mitotic spindles during the early development of seq urchin embryo, especially aiming the unequal division in the vegetable pole at 4th division. It was the first sign seen as cell differentiation and spincules were foumed only from the cells of michomere origin.
2) Mitotic arresters such as the alkylresorcinols and their derivatives, Macbecin I or heavy water (D_2O) were used to modulate microtubule assembly in the spindle with an aim to obtain necessitate informations on the microtubule control of anaphase chromosome movement.
3) Traces and image processings were attempted applying FITC-labeled tubulin of colcemid labeled with fluorescent dyes as the molecular probes to decipher the mechanism of in vitro assembly of microgubules. We also examined the applicability of Wiener's equation to analyze the spindle and kinetochore fibers., and probed its usefulness. Less
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