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Development of A Fiber Nano-scope with Molecular Trapping and its Application to the Analysis of Molecular Mechanism of Allergy

Research Project

Project/Area Number 09470495
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Physical pharmacy
Research InstitutionHIROSHIMA UNIVERSITY

Principal Investigator

MASUJIMA Tsutomu  Hiroshima University, Faculty of Medicine Professor, 医学部, 教授 (10136054)

Co-Investigator(Kenkyū-buntansha) TAMURA Atsushi  Hiroshima University, Faculty of Medicine Research Associate, 医学部, 助手 (30261225)
OZAWA Kouichiro  Hiroshima University, Faculty of Medicine Assistant Professor, 医学部, 講師 (10211822)
Project Period (FY) 1997 – 1998
Project Status Completed (Fiscal Year 1998)
Budget Amount *help
¥13,500,000 (Direct Cost: ¥13,500,000)
Fiscal Year 1998: ¥4,600,000 (Direct Cost: ¥4,600,000)
Fiscal Year 1997: ¥8,900,000 (Direct Cost: ¥8,900,000)
KeywordsMolecular Trapping / HPLC resin / MALDI-TOF Mass Spectrometry / Video Nanoscope / Optical Fiber / Cell Dynamism / Brownian Motion
Research Abstract

1. Development of Molecular Trapping Method and its Application to the Analysis of Allergy
Secreted molecules out of a mast cell model, RBL-2H3, were trapped by ion-exchange resins for HPLC in a single cell revel and its time course of secretion was monitored and analyzed by a fiber nano-scope. One species of the trapped molecules are identified to be histamine by matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF/MS). The molecular trapping probe of fiber type with ODS residues was also developed.
2. Development of A Fiber Nano-scope
A video-nanoscope with micro spot lighting with optical fiber light-guide was also developed. The Brownian movement of colloidal gold nano-particles was observed by this video scope and the movement of particleswas analyzed to evaluate the diameters of the particles. The dynamic movement of bound gold-labeled molecules was also observed by this new nano-scope.
3. Molecular Trapping and MALDI-TOF/MS Analysis
Not only the histamine but also the insulin which was secreted out of the beta pancreatic model cells was found to be detected and applied to the time course analysis of the beta pancreatic cell response.
4. Application to Another System
This method was also applied to the analysis of phagocytosis of neutrophils to clarify the sequential dynamism of the cells and clarified many new aspects of its behavior.

Report

(3 results)
  • 1998 Annual Research Report   Final Research Report Summary
  • 1997 Annual Research Report
  • Research Products

    (22 results)

All Other

All Publications (22 results)

  • [Publications] K.Ozawa: "Video-microscopy for analysis of molecualr dynamics in cells" J.Pharm.Biomed.Anal.15・7. 1483-1488 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] E.Suzaki: "Video-rate Dynamics of Exocytotic Events Associated With Phagocytosis in Neutrophils" Cell Motility and the Cytoskeleton. 38. 215-228 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] K.Ozawa: "Simultaneous analysis of membrane potential and calcium mobilization in a pancreatic β-cell line MIN6 by use of a double-probe imaging microscope system" Anal.Chim.Acta.365. 3-7 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] T.Masujima: "Trapping of the secreted molecules from a single cell using HPLC resin particles under video-microscope" Biol.Pharm.Bull.21・8. 886-888 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] A.Tamura: "Determination of Insulin Content in Pancreatic beta-Cell Line MIN6 Cells by Matrix-Assisted Laser Desorption Ionization Time-of-Flight Mass Spectrometry" Biol.Pharm.Bull.21・11. 1240-1241 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] C.Tao: "Pepsinogen Secretion:Coupling of Exocytosis Visualized by Video Microscopy and [Ca2+] in Single Cells" Am.J.Physiol.274. G1166-G1177 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] M.Suzuto: "Evaluation of Diameter of Colloidal Golds by Video-Analysis of its Brownian Motions" Anal.Chim.Acta.in press. (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] K.Ozawa, A.Tamura, K.Ikeda, E.Kawai, T.Kondo, Y.Fukano, S.Nomura, Y.Ishihara, and T.Masujima: "Video-microscopy for analysis of molecular dynamics in cells" J.Pharm.Biomed.Anal.15(7). 1483-1488 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] E.Suzaki, H.Kobayashi, Y.Kodama, T.Masujima, and S.Terakawa: "Video-rate Dynamics of Exocytotic Events Associated With Phagocytosis in Neutrophils" Cell Motility and the Cytoskeleton. 38. 215-228 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] K.Ozawa, Y.Fukano, T.Azuma, Y.Masumoto, H.Hayashi, A.Tamura, J.Miyazaki, and T.Masujima: "Simultaneous analysis of membrane potential and calcium mobilization in a pancreatic beta-cell line MIN6 by use of a double-probe imaging microscope system" Anal.Chim.Acta.365. 3-7 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] T.Masujima, K.Ikeda, K.Ozawa, A.Tamura, Y.Tada, H.Ohya, M.Suzuto: "Trapping of the secreted molecules from a single cell using HPLC resin particles under video-microscope" Biol.Pharm.Bull.21(8). 886-888 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] A.Tamura, T.Azuma, K.Ikeda, K.Ozawa, T.Masujima: "Determination of Insulin Content in Pancreatic beta-Cell Line MIN6 Cells aby Matrix-Assisted Laser Desorption Ionization Time-of-Flight Mass Spectrometry" Biol.Pharm.Bull.21(11). 1240-1241 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] C.Tao, M.Yamamoto, H.Mieno, M.Inoue, T.Masujima, G.Kajiyama: "Pepsinogen Secretion : Coupling of Exocytosis Visualized by Video Microscopy and [Ca2+] in Single Cells" Am.J.Physiol.274. G1166-G1177 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] M.Suzuto, A.Nakamura, Y.Ishihara, K.Ikeda, A.Tamura, T.Masujima: "Evaluation of Diameter of Colloidal Golds by Video-Analysis of its Brownian Motions" Anal.Chim.Acta. (in press). (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] C.Tao: "Pepsinogen Secretion:Coupling of Exocytosis Visualized by Video Microscopy and [Ca2+]in Single Cells" Am.J.Physiol.274. G1166-G1177 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] K.Ozawa: "Simultaneous analysis of membrane potential and calcium mobilization in a pancreatic β-cell line MIN6 by use of a double-probe imaging microscope system" Anal.Chim.Acta.365. 3-7 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] T.Masujima: "Trapping of the secreted molecules from a single cell using HPLC resin particles under video-microscope" Biol.Pharm.Bull.21・8. 886-888 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] A.Tamura: "Determination of Insulin Content in Pancreatic beta-Cell Line MIN6 Cells by Matrix-Assisted Laser Desorption Ionization Time-of Flight Mass Spectrometry" Biol.Pharm.Bull.21・11. 1240-1241 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] M.Suzuto: "Evaluation of Diameter of Colloidal Golds by Video-Analysis of its Brownian Motions" Anal.Chim.Acta. (in press). (1999)

    • Related Report
      1998 Annual Research Report
  • [Publications] K.Ozawa: "Video-microscopy for analysis of molecular dynamics in cells" J.Pharm.Biomed.Anal.15・7. 1483-1488 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] A.Tamura: "Direct observation of sample trpping process onto internal-surface reversed phase presample trapping process onto internal-surface reversed phase pre-column" Anal.Chim.Acta.(in press). (1998)

    • Related Report
      1997 Annual Research Report
  • [Publications] T.Masujima: "Trapping of the secreted molecules from a single cell using HPLC resin particles under videomicroscope" Biol,Pharm.Bull.(in press). (1998)

    • Related Report
      1997 Annual Research Report

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Published: 1997-04-01   Modified: 2016-04-21  

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