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1991 Fiscal Year Final Research Report Summary

Development of analytical color fluorescence electron microscope (ACFEM) and its application to the medical and biological fields

Research Project

Project/Area Number 01870002
Research Category

Grant-in-Aid for Developmental Scientific Research

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

Principal Investigator

OGAWA Kazuo  Department of Anatomy, Kyoto University, Professor, 医学部, 教授 (20077556)

Co-Investigator(Kenkyū-buntansha) SAKAI Masahiro  Department of Anatomy, Kyoto University, Instructor, 医学部, 助手 (40183363)
FUJIMOTO Kazushi  Department of Anatomy, Kyoto University, Lecturer, 医学部, 講師 (50159125)
FUJIMOTO Toyoshi  Department of Anatomy, Kyoto University, Associate Professor, 医学部, 助教授 (50115929)
Project Period (FY) 1989 – 1991
KeywordsACFEM / Cathodoluminescence (CL) / Analytical EM / CL analysis / Cholesterol ester / Vitamin A ester
Research Abstract

In order to establish a new method for an electron microscopic cytochemical analysis, we developed an analytical color fluorescence electron microscope(ACFEM)and applied this method to the biomedical research. On the basis of a high resolution scanning electron microscope, the ACFEM, equipped with a cathodoluminescence(CL)-collecting mirror and a spectral multichannel analytical system, enables us to obtain color CL images, monochromatic CL images at any given wavelength, and conventional SEN images, as well as CL spectrum when an electron beam is scanning over the surface of specimens.
Using the ACFEM, we observed CL from a varieties of organs and tissues without fluorescent staining to analyze the substances which emit CL. CL was found to be emitted from lipid droplets(LD)in biological specimens with a two-peak pattern spectrum at 320 nu and 430-450 na respectively. It was proved that the 320 nm-CL was derived from cholesterol esters, because 320 nu-CL, which was usually detected in t … More he LD of adrenal cortex, corpus luteum, theca interna and granulosa cells of the rats, disappeared when the animals were treated with hypocholesterolemic agents, and showed an increase of intensity after hypophysectomy. Similarly, the 430-450 na-CL was demonstrated to be derived from a mixture of lipids at least including vitamin A ester by the research dealt with retina and testes of the rats. So far, it was impossible to analyze the components in a single lipid droplet by means of cytochemistry for the reason of lacking antibodies with high specialities against lipids, such as steroids and retinoids. This study methodologically dissolved this problem.
Moreover, we also observed that FITC-IgG emits CL under the ACF24. This fact indicates the possibility to directly introduce fluorescent antibody method to electron microscopy. Recently, the study on evaluating diamonds and developing new photo sensitive materials have revealed that the ACFEM could be very useful in the industrial research fields, also. Less

  • Research Products

    (19 results)

All Other

All Publications (19 results)

  • [Publications] T.Nakano,T.Fujimoto,H.Koike,K.Ogawa: "Application of analytical color fluorescence electron microscopy to biomedical field:I.Vitamin A ester in rat retina." Acta histochemica et cytochemica. 23. 753-767 (1990)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] T.Nakano,T.Fujimoto,H.Koike,K.Ogawa: "Application of analytical color fluorescence electron microscopy to biomedical field:II.Cholesterol ester in rat adrenal cortex." Acta histochemica et cytochemica. 23. 769-780 (1990)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] A.Nitro,T.Nakano,G.Ning,K.Ogawa: "Effect of hypophysectomy on rat adrenocortical cells studied by analytical color fluorescence electron microscopy." Acta Histochemica et Cytochemca. 24. 99-101 (1991)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 小川 和朗: "分析カラ-蛍光電子顕微鏡の開発" 学術月報. (1992)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] K.Ogawa,T.Nakano,G.Ning,T.Fujimoto,K.Nishimura,H.Koike: "International Symposium on Electron Microscopy" World Scientific Publishing Co.Ltd.Ed.by k.H.Kuo and J.E.Yao, 188-207 (1991)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] T.Nakano,G.Ning: "Electron Microscopic Cytochemistry and Immunocytochemistry in Biomedicine" CRC Press,Florida Ed.by K.Ogawa and T.Barka, (1992)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Nakano, T.: "Color fluorescence electron microscopy and cathodoluminescence spectra analysis." Journal of Histochemistry and Cytochemistry. 36(7a). 915 (1988)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Ogawa, K.: "Development of color fluorescence electron microscope and its application to the biological specimens." Verhaudlungen der anatomischen Gesellsehaft 82 (Anatomischer. Anzeiger. Suppl. 164). 303-304 (1989)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Ogawa, K.: "Analytical color fluorescence electron microscopy." Journal of Electron Microscopy. 39. 284 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Nakano, T.: "application of analytical color fluorescence electron microscopy to biological field : I. Vitamin A ester in rat retina." Acta Cytochemica ct Histochemica. 23(6). 753-767 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Nakano, T.: "Application of analytical color fluorescence electron microscopy to biological field : II. Cholesterol ester in rat adrenal cortex." Acta Cytochemica et Histochemica. 23(6). 769-780 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Ning, G.: "Cathodoluminescence (CL) from lipid droplets (LD) of various follicles of rat by analytical color fluorescence electron microscopy." Journal of Histochemistry and Cytochemistry. 36(7a). 915 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Mitro, A.: "Effect of hypophysectomy on rat adrenocortical cells studied by analytical color fluorescence electron microscopy." Acta Cytochemica et Histochemica. 24(1). 99-101 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Ning, G.: "Detection of lipid droplets in rat seminiferous tubules by analytical color fluorescence electron microscopy." Proceedings of the lat. China - Japan Joint Histochemistry and Cytochemistry Seminar, ed. by S. Fujita, M. Ai, K. Ogawa and Y. J. Piao, Guangzhou. 9-10 (1989)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Ogawa, K.: "Recent development of analytical color fluorescence electron microscopy : application to biomedical and industrial fields." International Symposium on Electron Microscopy, ed. by K. Kuo and J. Yao, World Scientific, Singapore. 188-207 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Ning, G.: "Analysis of cathodoluminescence emitted form lipid droplets in corpora lutea of estrus cyclic and pregnant rat by analytical color fluorescence electron." Proceedings of the 12th International Congress for Electron Microscopy, ed. by L. D. Peachey and D. B. Williams, San Francisco Press. 2. 768-769 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Nakano, T.: "Analysis of rat adrenocortical cells by analytical color fluorescence electron microscopy." Proceedings of the 12th International Congress for Electron Microscopy, ed. by L. D. Peachey and D. B. Williams, San Francisco Press. 2. 770-771 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Nakano, T: "Analytical color fluorescence electron microscopy." Electron Microscopic Cytochemistry and Immunocytochemistry in Biomedicine, ed. by K. Ogawa and T. Barka, CRC Press, Florida. (1992)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Ning, G.: "Cathodoluminescence in rat ovary : a study by analytical color fluorescence electron microscopy." Recent Development of Electron Microscopy 1991, Proceedings of the 6th Japanese - Chinese Electron Microscopy Seminar, ed. by H. Hashimoto, K. H. Kuo, K. Lee and K. Ogawa. Okayama. (1992)

    • Description
      「研究成果報告書概要(欧文)」より

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Published: 1993-03-16  

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