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Application of rapid-frozen substitution fixation method to immunohistochemistry

Research Project

Project/Area Number 61870001
Research Category

Grant-in-Aid for Developmental Scientific Research

Allocation TypeSingle-year Grants
Research Field General anatomy (including Histology/Embryology)
Research InstitutionKeio University

Principal Investigator

YASUDA Kenjiro  Professor, Keio University School of Medicine, 医学部, 教授 (90050327)

Co-Investigator(Kenkyū-buntansha) ICHIKAWA Atsushi  Professor, Yokohama City University School of Medicine, 医学部, 教授 (50045910)
Project Period (FY) 1986 – 1987
Project Status Completed (Fiscal Year 1987)
Budget Amount *help
¥8,100,000 (Direct Cost: ¥8,100,000)
Fiscal Year 1987: ¥1,600,000 (Direct Cost: ¥1,600,000)
Fiscal Year 1986: ¥6,500,000 (Direct Cost: ¥6,500,000)
Keywordsrapid-frozen substitution fixation method / immunohistochemistry / parotid gland / pancreas / amylase / ヘリウム / 耳下腺 / 膵蔵 / ラット
Research Abstract

A rapid-freezing substitution technique is a quite useful tool to study the ultrastructural features of cells in a more life-like state. However, there remained several difficulties in applying the technique to immunoelectron microscopy. In this study, we tried to overcome those difficulties to perform immunolabelling in well- preserved ultrastructure.
Frozen rat parotid gland and exocrine pancreas tissue blocks were quenched by smaching against a copper block surface which was pre-cooled with liquid helium. The tissue blocks were freeze-substituted in either of the following media; osmium in acetone, paraformaldelhyde in acetone, and glutaraldehyde in acetone. Paraformaldehyde in acetone and glutaraldehyde in acetone were prepared with our new distillation device. After substitution, trimmed materials were embedded. Ultrathin sections were immunostained with rabbit anti-rat-amylase antibodies and protein A/colloidal gold.
High labelling density was obtained from the materials which were fixed by freeze- substitution with glutaraldehyde in acetone or paraformaldehyde in acetone, embedded in Araldite 6005. In materials substituted with Os04 in acetone, embedded in an epoxy resin mixture and etched with sodium metaperiodate solution, labelling density was lowr than the former. In Lowicryl K4M-embedded materials labelling was the ghighest among the mataerials examined so far.
The results in this study present an useful and preactical information in the application of immunocytochemistry to rapid-frozen substitution fixed tissues.

Report

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

    (33 results)

All Other

All Publications (33 results)

  • [Publications] Kenjiro Yasuda;Shuji Yamashita;Sadakazu Aiso;Masahide Shiozawa and Toshiko Komatsu: Acta histochemica et cytochemica. 19(5). 589-600 (1986)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] Masahide Shiozawa;Sadakazu Aiso; Osamu Hoshiai;Hiromu Tahara;Shuju Yamashita and Kenjiro Yasuda: Okahimas Folia Anatomica Japonica. 63(4). 209-222 (1986)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] 安田健次郎: 日本医師会雑誌. 95(2). 239-240 (1986)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] Atsushi Ichidawa and Misao Ichikawa: Journal of Electron Mecroscopy. 35(SUPPL). 1979-1980 (1986)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] Kenjiro Yasuda: Journal of Electoron Microscopy. 35(SUPPL). 2019-2020 (1986)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] Misao Ichikawa;Katsunori Sasaki and Atsushi Ichikawa: Journal of Electron Microscopy. 35(SUPPL). 2301-2302 (1986)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] Toshiko Komatsu; Sjiji Yamashita; Kenjiro Yasuda: Acta Histochemica et Cytochemica. 20(2). 163-176 (1987)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] 安田健次郎: 医学のあゆみ. 142(11). 789-792 (1987)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] Kenjiro Yasuda; Sadakazu Aiso; Takashi Nagatani and Mitsuhiko Yamada: Acta Histochemica et Cytochemica. 20(3). 371-372 (1987)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] 市川厚: 電子顕微鏡. 22(2). 124-128 (1987)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] Atsusi Ichicawa; Kejiro Yasuda; Misao Ichidawa; Shuji Yamashita; Sadakazu Aiso and Masahide Shiozawa: Journal of Histochemistry and Cytochemistry.

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] 山下修二,安田健次郎: "組織細胞化学1986(モノクローナル抗体の作製法)" 学際企画株式会社, 282 (1986)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] 安田健次郎: "電子顕微鏡生物試料作製法(〓疫化学的方法,)" 丸善株式会社, 318 (1986)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] 安田健次郎: "膵臓外分泌部(人体組織学:消化器" 朝倉書店, 527 (1987)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] Kenjiro Yasuda, Shuji Yamashita, Sadakazu Aiso, Masahide Shiozawa and Toshiko Komatsu: "Immunohistochemical study of -glutamyl transpeptidase with monoclonal antibodies. I. Preparation and characterization of monoclonal antibodies to -glutamyl transpeptidase" Acta histochemica et cytochemica. 19. 589-600 (1986)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] Masahide Shozawa, Sadakazu Aiso, Osamu Hoshiai, Hiromu Tahara, Shuji Yamashita and Kenjiro Yasuda: "Localization of gamma-glutamyl transpeptidase in the proliferative state of liver. An enzyme histochemical and immunohistochemical study" Okajimas Folia Anatomica Japonica. 63. 209-222 (1986)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] Kenjiro Yasuda: "Monoclonal antibodies--anfundamental study for clinical applications (in japanese)" Journal of Japan Medical Association. 95. 239-240 (1986)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] Atsushi Ichikawa and Misao Ichikawa: "Application of cryofixation to immunocytochemistry" Journal of Electron Microscopy. 35 suppl.1979-1980 (1986)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] Kenjiro Yasuda: "Immunoelectron microscopy with colloidal gold on thin sections of rappid-frozen substitution fixed specimen" Journal of Electron Microscopy. 35 suppl.2019-2020 (1986)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] Misao Ichikawa, Katsunori Sasaki and Atsushi Ichikawa: "Immunocytochemical localization of amylase in gerbil salivery glandular cells processed by rapid freezing and freeze-substitution fixation" Journal of Electron Microscopy. 35 suppl.2301-2302 (1986)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] Toshiko Komatsu, Shuji Yamashita and Kenjiro Yasuda: "Preparation of monoclonal antibody to rabbit kidney gamma-glutamyl transpeptidase and its cross-reactivity analysis in immunohistochemistry" Acta histochemica et cytochemcia. 20. 163-176 (1987)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] Kenjiro Yasuda: "ARIICLE: Monoclonal antibodies and immunohistochemistry (in japanese)" Igakuno Ayumi. 142. 789-792 (1987)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] Kenjiro Yasuda, Sadakazu Aiso, Takashi Nagatani and Mitsuhiko Yamada: "A study to demonstrate antibody molecules on the sections in immunoelectron microscopy" Acta histochemica et cytochemica. 20. 371-372 (1987)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] Atsushi Ichikawa: "AR@iCLE: Application of cryofixation to immunohistochemistry" Electron Microscopy. 22. 124-128 (1987)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] Atsushi Ichikawa, Kenjiro Yasuda, Misao Ichikawa, Shuji Yamashita, Sadakazu Aiso and Masshide Shiozawa: "An application of cryofixation to immunohistochemistry" Journal of Histochemistry and Cytochemistry. in press. (1987)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] Shuji Yamashita and Kenjiro Yasuda: Gakusai Kikaku. Preparation of monoclonal antibodies (in Histocytochemistry 1986), 175-194 (282) (1986)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] Kenjiro Yasuda: Maruzen. Immunochemical methods (in Preparation of biological specimen for electron microscopy), 219-236 (318) (1986)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] Kenjiro Yasuda: Asakura Shoten. Exocrine pancreas ( in Human histology), 453-459 (527) (1987)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] Kenjiro Yasuda;Shuji Yamashita;Sadakazu Aiso;Masahide Shiozawa;Toshiko Komatsu: Acta histochemica et cytochemica. 19(5). 589-600 (1986)

    • Related Report
      1986 Annual Research Report
  • [Publications] Masahide Shiozawa;Sadakazu Aiso;Osamu Hoshiai Hiromu Tahara;Shuji Yamashita;Kenjiro Yasuda: Okajimas Folia Anatomica Japonica. 63(4). 209-222 (1986)

    • Related Report
      1986 Annual Research Report
  • [Publications] 安田健次郎: 日本医師会雑誌. 95(2). 239-240 (1986)

    • Related Report
      1986 Annual Research Report
  • [Publications] 山下修二,安田健次郎: "組織細胞化学1986(モノクローナル抗体の作製法)" 学際企画株式会社, 282 (1986)

    • Related Report
      1986 Annual Research Report
  • [Publications] 安田健次郎: "電子顕微鏡生物試料作製法(免疫化学的方法)" 丸善株式会社, 318 (1986)

    • Related Report
      1986 Annual Research Report

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

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