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Three-dimensional Image Analysis of Muscle Cell Structures

Research Project

Project/Area Number 04044034
Research Category

Grant-in-Aid for international Scientific Research

Allocation TypeSingle-year Grants
SectionJoint Research
Research InstitutionGunma University

Principal Investigator

ISHIKAWA Harunori  Gunma University School of Medicine, 医学部, 教授 (90010058)

Co-Investigator(Kenkyū-buntansha) PEACHEY Lee d.  University of Pennsylvania, Department of Biology, 生物学科, 教授
L D Peachey  ペンシルバニア大学, 生物学教室, 教授
藤巻 昇  群馬大学, 医学部, 講師 (10008261)
Project Period (FY) 1992 – 1994
Project Status Completed (Fiscal Year 1994)
Budget Amount *help
¥7,200,000 (Direct Cost: ¥7,200,000)
Fiscal Year 1994: ¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 1993: ¥2,300,000 (Direct Cost: ¥2,300,000)
Fiscal Year 1992: ¥3,400,000 (Direct Cost: ¥3,400,000)
KeywordsThree-dimensional image analysis / Striated muscle / T-system / Electron Microscopy / Confocal laser microscopy / LM-EM correlation / Myofibril / Membranous system / 横細管系 / 選択的染色
Research Abstract

1.Three-Dimensional Image Analysis of Membranous Systems in Skeletal Muscle
The T-system in frog skeletal muscle fibers was selectively stained with the Golgi silver impregnation method followed by specimen preparation for electron microscopy. We found that the muscle fibers in which T-system was selectively stained appeared transparent, while those with sarcoplasmic reticulum staining were pink and those with no membranous staining were in black under reflected light. Thus, the muscle fibers with T-system staining were easily identified under the light microscope.
Sections of 1-3mum thick were cut from the Golgi-stained muscle tissue and examined with the intermediate voltage electron microscope. Such stereo-pair micrographs were taken to be used for development of computer programs for three-dimensional image analysis.
When Golgi-stained muscle fibers were examined under the confocal laser microscope with reflection mode, the silver-stained T-system was clearly visualized in three dimen … More sions. Such confocal images were found to be useful for studies of the overall distribution of the T-system.
2.Correlated Light and Electron Microscopy of Muscle Cells.
We developed the method to correlate the fluorescence and immunofluorescence findings to ultrastructures observed by electron microscopy. For cultured cells, cardiac muscle cells from chick embyros were cultured on formval-filmed gold EM grids, stained fluorescently, and examined by confocal laser microscopy. The same samples were then fixed, dehydrated, and critical-point-dried for whole-mount cell specimen preparations, which were then examined three-dimensionally with the intermediate voltage electron microscope. In this method, we could successfully correlate the fluorescence images with ultrastructures in the same cells. The similar correlation was also made for tissue sections. Cryo-stat sections were stained for immunofluorescence using anti-dystrophin antibody and examined by confocal laser microscopy. Serial optical section images of about 0.5 mum thick were reconstructed three-dimensionally to corelate in a point-to-point manner with stereo-pair images obtained from electron microscopy. Less

Report

(3 results)
  • 1994 Final Research Report Summary
  • 1993 Annual Research Report
  • 1992 Annual Research Report
  • Research Products

    (30 results)

All Other

All Publications (30 results)

  • [Publications] Masuda, T.: "Confocal laser microscopy of dystrophin localization in guinea pig skeletal muscle fibers." J. Cell Biol.119. 534-548 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Ishikawa, H.: "Immunohistochemical localization of the proteins associated with brain N-type calcium channels." Ann. New York Acad. Sci.707. 376-378 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Fujimaki, N.: "Light-microscopic identification of Golgi staining patterns in embedded blocks of muscle tissue prior for-" Proc. 51st Ann. Meet. Microsc. Soc. Amer.344-345 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Murakami, T.: "Confocal interference reflection microscopy of cultured cells." Bioimages. 1. 1-10 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Fujimaki, N.: "Three-dimensional visualization of the T-system in fixed and embedded frog skeletal muscle fibers by con-" Bioimages. 1. 167-174 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Katoh, K.: "Organization of the cytoskeleton during cellularization in Drosophila melanogaster embryos; Confocal laser" Bioimages. 2. 59-69 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] 石川春律(共著): "実験生物物理" 丸善, 243 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] 石川春律(共著): "組織細胞化学 1994" 学際企画, 249 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] 石川春律(共編著): "現代病理学大系 4" 中山書店, 387 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] 石川春律(共著): "共焦点レーザー顕微鏡の医学・生物学への応用" 学際企画(印刷中), (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Masuda, T.: "Confocal laser microscopy fo dystrophin localization in guinea pig skeletal muscle fibers." J.Cell Biol.119. 543-548 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Ishikawa, H.: "Immunohistochemical localization of the proteins associated with brain N-type calcium channels." Ann.New York Acad.Sci.707. 376-378 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Fujimaki, N.: "Light-microscopic identification of Golgi staining patterns in embedded blocks of muscle tissue prior for -" Proc.51st Ann.Meet.Microsc.Soc.Amer.344-345 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Murakami, T.: "Confocal interference reflection microscopy of cultured cells." Bioimages. 1. 1-10 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Fujimaki, N.: "Three-dimensional visualization of the T-system in fixed and embedded frog skeletal muscle fibers by confocal laser-" Bioimages. 1. 167-174 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Katoh, K.: "Organization of the cytoskeleton during cellularization in Drosophila melanogaster embryos ; Confocal laser -" 2. 59-69 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Murakami,T.: "Confocal interference reflection microscopy of cultured cells." Bioimages. 1. 1-10 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] Ono,M.: "Association of the actin cytoskeleton with glass-adherent proteins in mouse peritoneal macrophages." Biology of the Cell. 77. 219-230 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] Ishikawa,H.: "Immunohistochemical localization of the proteins associated with brain N-type calcium channels." Annals of the New York Academy of Sciences. 707. 376-378 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] Fujimaki,N.: "Three-dimensional visualization of the T-system in fixed and embedded frog skeletal muscle fibers by confocal laser reflection microscopy." Bioimages. 1. 167-174 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] Fujimaki,N.: "Light-microscopic identification of Golgi staining patterns in embedded blocks of muscle tissue prior for electron microscopy." Proc.51stAnn.Meet.Microsc.Soc.Amer.344-345 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] Murakami,T.: "Confocal microscopic 3D visualization of membranous organelles in living cultured fibroblasts." Bioimages. 2. 21-28 (1994)

    • Related Report
      1993 Annual Research Report
  • [Publications] MASUDA,T.: "Confocal laser microscopy of dystrophin localization in guinea pig skeletal muscle fibers." J.Cell Biol.119. 543-548 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] MURAKAMI,T.: "Confocal interference microscopy of cultured cells." Bioimages. 1. 1-10 (1993)

    • Related Report
      1992 Annual Research Report
  • [Publications] NAKAZAWA,E.: "The association of microtubules with the plasmalemma in epidermal tendon cells of the river crab." Biol.Cell. 75. 111-119 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] TOMITA,M.: "Identification of novel adhesion proteins in mouse peritoneal macrophages." Biol.Cell. 76. 103-109 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] ONO,M.: "Association of the actin cytoskeleton with glass-adherent proteins in mouse peritoneal macrophages." Biol.Cell. 77. IN PRESS (1993)

    • Related Report
      1992 Annual Research Report
  • [Publications] FUJIMAKI,N.: "Light microscopic identification of Golgi staining patterns in embedded blocks of muscle tissues prior to sectioning for electron microscopy" Proc.51st Annu.Meet.Microsc.Soc.Amer.IN PRESS (1993)

    • Related Report
      1992 Annual Research Report
  • [Publications] 石川 春律(分担): "組識細胞化学1992" 日本組識細胞化学会, 174 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] 村上 徹(分担): "細胞生物学1992" 中外医学社, 335 (1992)

    • Related Report
      1992 Annual Research Report

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Published: 1992-04-01   Modified: 2016-04-21  

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