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Image analysis of weakly birefringnet cytoskeletons recorded by high resolution DIC-Pol microscope and a sensitive time-lapsc VTR combined with image analyzer.

Research Project

Project/Area Number 60480020
Research Category

Grant-in-Aid for General Scientific Research (B)

Allocation TypeSingle-year Grants
Research Field 動物発生・生理学
Research InstitutionSchool of Science, Nagoya University

Principal Investigator

SATO Hidemi  Sugashima Marine Biological Loboratory, Nagoya University, 理学部, 教授 (40109260)

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)
Project Period (FY) 1985 – 1987
Project Status Completed (Fiscal Year 1987)
Budget Amount *help
¥5,300,000 (Direct Cost: ¥5,300,000)
Fiscal Year 1987: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 1986: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1985: ¥3,700,000 (Direct Cost: ¥3,700,000)
KeywordsWeak Birefringence / Cytoskeleton / Mitotic Spindle / Microtubules / 高感度VTR / 画像解折 / 高解像ノマルスキー効果
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

Report

(2 results)
  • 1987 Final Research Report Summary
  • 1986 Annual Research Report
  • Research Products

    (23 results)

All Other

All Publications (23 results)

  • [Publications] Itoh, T. J. ;Sato, H. & Kodayashi, A.: Zoological Science. 4. 265-276 (1987)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] 佐藤英美: 生体の科学. 38;No. 5. 467-475 (1987)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] Obata, S. & Kuroda, H.: Developmental Biology. 122. 432-438 (1987)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] Hiratsuka, T. & Kato, T.: Journal of Biochemistry. 262. 6138-6322 (1987)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] Sato, H. ;Kobayashi, A. & Itoh, T. J.: Critical Reviw of Anatomy. CRC Press Inc.(1988)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] Sato, H. ;Sato, Y. ;Bajer, A. & Mole-Bajer: Photoplasma, Memorial issue for Prof. N. Kamiya.(1988)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] Itoh,T.J., Sato,H & Kobayashi,A.: "Examination of spindle structures modified by T-1, the mitic arrester." Zoological Science. 4. 265-276 (1987)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] Obata,S. & Kuroda,H.: "The second component of fertilization potential in sea urchin (Pseudocentrotus depressus) eggs involves both N_a^+ and K^+" permeability. Developmental Biology. 122. 432-438 (1987)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] Hiratsuka,T. & Kato,T.: "A Fluorescent analog of colcemid, N-(7-nitrobenz-2-oxa-1,3- diazol-4-yl)-colcemid, as a probe for the colcemid binding sistes of tubulin and microtubules. Jounal of Biochemistry" 262. 6318-6322 (1987)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] Sata,H., Kobayashi,A. & Itoh,T.J.: "Molecular basis of physical and chemical probes for spindle assembly." Critical Review of Anatomy, CRC.(1988)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] Sato,H., Sato,Y.,J. Mole-Bajer, & Bajer,A.S.: "Application of Wiener's equation to analyze form birefringence in dividng endosperm cell, Haemanthus katherinae." Protoplasma: Memorial issue for Professor Kamiya,N.(1988)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] Takane Ohara;Hidemi Sato: Development,Growth and Differentiation. 28. 125-135 (1986)

    • Related Report
      1986 Annual Research Report
  • [Publications] Tomohiko J.Itoh;Hidemi Sato;Akio Kobayashi: Zoological Science. 3. 225-264 (1986)

    • Related Report
      1986 Annual Research Report
  • [Publications] Takane Ohara;Hidemi Sato: Development,Growth and Differentiation. 28. 369-373 (1986)

    • Related Report
      1986 Annual Research Report
  • [Publications] Andrew,S.Bajer;Hidemi Sato;J.Mole-Bajer: Cell Structure and Function. 11. 317-330 (1986)

    • Related Report
      1986 Annual Research Report
  • [Publications] 佐藤英美: 生体の科学特集「細胞生物学実験マニュアル」. 37. 313-315 (1986)

    • Related Report
      1986 Annual Research Report
  • [Publications] Tomohiko J.Itoh,Hidemi Sato and Akio Kobayashi: Zoological Science. 4. (1987)

    • Related Report
      1986 Annual Research Report
  • [Publications] 佐藤英美: 細胞工学. 6. (1987)

    • Related Report
      1986 Annual Research Report
  • [Publications] Shuichi Obata;Hideyo Kuroda: Developmental Biology. 124. (1987)

    • Related Report
      1986 Annual Research Report
  • [Publications] Toshiaki Hiratsuka and Toyoki Kato: Journal of Biological Chemistry. (1987)

    • Related Report
      1986 Annual Research Report
  • [Publications] Toyoki Kato and Toshiaki Hiratsuka: European Journal of Biochemistry.

    • Related Report
      1986 Annual Research Report
  • [Publications] 佐藤幸子: 松蔭女子学院大学・短期大学自然科学研究紀要. (1987)

    • Related Report
      1986 Annual Research Report
  • [Publications] 佐藤英美,丸山・矢原 編: ""複屈折法"続生化学実験講座6「細胞骨格の構造と機能」" 東京化学同人, 12 (1986)

    • Related Report
      1986 Annual Research Report

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Published: 1987-03-31   Modified: 2016-04-21  

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