• Search Research Projects
  • Search Researchers
  • How to Use
  1. Back to previous page

Highly-Selective Determination of Chemical Sepecies in Cells with 3D Fluorescence Microscopy

Research Project

Project/Area Number 10555294
Research Category

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

Allocation TypeSingle-year Grants
Section展開研究
Research Field 工業分析化学
Research InstitutionKYUSHU UNIVERSITY

Principal Investigator

HARATA Akira  KYUSHU UNIVERSITY, Faculty of Matrl. Sci., Ass. Prof., 大学院・総合理工学研究院, 助教授 (90222231)

Co-Investigator(Kenkyū-buntansha) 小川 禎一郎  九州大学, 大学院・総合理工学研究科, 教授 (50028156)
Project Period (FY) 1998 – 2000
Project Status Completed (Fiscal Year 2000)
Budget Amount *help
¥13,100,000 (Direct Cost: ¥13,100,000)
Fiscal Year 2000: ¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 1999: ¥1,800,000 (Direct Cost: ¥1,800,000)
Fiscal Year 1998: ¥9,900,000 (Direct Cost: ¥9,900,000)
KeywordsLaser-Induced Fluorescence / Fluorescence Microscope / Spectroscopic Image / Three Dimensional Analysis / Highly-Selective Determination / Biological Substances / Ion Exchange Resin / Particle Counting / 細胞内物質 / ウイルス計数
Research Abstract

A fluorescence method is the one of the most sensitive detection tools for chemical species but determination of amounts of functional substances in a biological cell for diagnostics always faces many practical problems : Large backgrounds caused by nonspecifically adsorbed-fluorescence probe and organic substances makes high sensitive detection impossible ; Light scattering from the cell walls disturbs reproducibility ; and so on. This study aims to solve these problems and get practical guidelines for a fluorescence method of cell-based diagnostics.
1) A three-dimensional fluorescence micro-spectrometer is composed. A laser source or monochromated halogen-lamp light source, a microscope and an imaging spectrometer are combined to observe spectral images of microscopic substances.
2) Discrimination of individual microparticles is demonstrated with the fluorescence micro-spectrometer. Different types of fluorescence dyes are adsorbed on different microparticles, respectively. They are made of polystyrene or ion-exchange resin, and their diameters are from 3 to 11 μm. After mixing, they are used as test samples. It was found that each microparticle was distinguishable even if their fluorescence emission spectra have the same shapes with a difference in their peak wavelengths of only 5-10 nm. Particle discrimination under a high background condition was demonstrated as well.
3) Detection limit for adsorbed dyes on one microparticle was determined to be as low as 10^<-18> mol/particle.
4) Based on the experimental results, a guideline for fluorescence determination of chemical species in micro-substances was proposed where fluorescence intensity, spectra and background levels are evaluated for the diagnostics.

Report

(4 results)
  • 2000 Annual Research Report   Final Research Report Summary
  • 1999 Annual Research Report
  • 1998 Annual Research Report
  • Research Products

    (17 results)

All Other

All Publications (17 results)

  • [Publications] X.Zheng,A.Harata,T.Ogawa: "Photon Bursts from Low Surface-Density Dioctadecyl-Rhodamine B Molecules at the Air-Water Surface Observed with a Confocal Fluorescence Photon-Counting Microscope"Chemical Physics Letters. 316. 6-12 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] A.Harata,N.Yamaguchi: "Photothermal Lensing Signal Enhancement by Transient Absorption of Photoexcited States in Liquid Solutions"Analytical Sciences. 16. 743-749 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] X.Zheng,A.Harata,T.Ogawa: "Confocal Fluorescence Microscope Studies of the Adsorptive Behavior of Dioctadecyl-Rhodamine B Molecules at a Cyclohexane-Water Interface"Analytical Sciences. 17. 131-135 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] X.Zhang, A.Harata, T.Ogawa: "Fluorescence Photon Bursts from Low Surface-Density Dioctadecyl-Rhodamine B Molecules at the Air-Water Surface Observed with a Confocal Fluorescence Photon-Counting Microscope."Chemical Physics Letters. 316. 6-12 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] A.Harata, N.Yamaguchi: "Photothermal Lensing Signal Enhancement by Transient Absorption of Photoexcited States in Liquid Solutions."Analytical Scienses. 16. 743-749 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] X.Zheng, A.Harata: "Confocal Fluorescence Microscope Studies of the Adsorptive Behavior of Dioctadecyl-Rhodamine B Molecules at a Cyclohexane-Water Interface"Analytical Scienses. 17. 131-135 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Valeria TSUKANOVA,Akira HARATA,Teiichiro OGAWA: "Orientational Arrangement of Long-Chain Fluorescein Molecules within the Monolayer at the Air/Water Interface Studied by the SHG Technique"Langmuir. 16・3. 1167-1171 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Valeria TSUKANOVA,Akira HARATA and Teiichiro OGAWA: "Second-Harmonic Probe of Pressure-Induced Structure Ordering within Long-Chain Fluorescence Monomer at the Air/Water Interface"Journal of Chemical Physics. 104・32. 7707-7712 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Akira HARATA and Naonori YAMAGUCHI: "Photothermal Lensing Signal Enhancement by Transient Absorption of Photoexcited States in Liquid Solutions"Analytical Sciences. 16. 743-749 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Xue-Ying ZHENG and Akira HARATA: "Confocal Fluorescence Microscope Studies of the Adsorptive Behavior of Dioctadecyl-Rhodamine B Molecules at a Cyclohexane-Water Interface"Analytical Sciences. 17. 131-135 (2001)

    • Related Report
      2000 Annual Research Report
  • [Publications] X..-Y.Zheng,A.Harata,T.Ogawa: "Fluorescence Photon Bursts from Low Surface-Density Dioctadecy1-Rhodamine B Molecules at the Air-Water Surface Observed with a Confacal Fluorescence Photon-Counting Microscope"Chemical Physice Letters. 316. 6-12 (2000)

    • Related Report
      1999 Annual Research Report
  • [Publications] M.N.Slyadnev,T.Inoue,A.Harata,T.Ogawa: "Investigation of Water Surface by Laser Induced Fluorescence Microscopy using Cyanine and Rhodamine Dye"Colloids and Surfaces A. 164・2/3. 155-162 (2000)

    • Related Report
      1999 Annual Research Report
  • [Publications] Y.-Q.Li,M.N.Slyadonev,T.Inoue,A.Harata,T.Ogawa: "Spectral Fluctuation and Heterogeneous Distribution of Porphine on a Water Surface"Langmuir. 15・9. 3035-3037 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Y.-Q.Li,T.Inoue,A.Harata,T.Ogawa: "A Stategy to Probe Air-Liquid Interfaces by Confocal Fluorescent Microscoy"Instrumental Science and Technology. 27・3. 159-166 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] O.N.Slyadneva,M.N.Slyadnev,V.M.Tsukanova,T.Inoue,A.Harata,T.Ogawa: "Orientation and Aggregation Behavior of Rhodamine Dye in Insoluble Film at the Air-Water Interface under Compression.Second Harmonic Generation and Spectroscopic Studies"Langmuir. 15・25. 8651-8658 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Y.-Q. Li, Sakaki, T. Inoue, A. Harata, T. Ogawa: "Spatial Imaging Identification in a Fiber-Bundle Confocul Fluorescence Microspectrometer" Applied Spectroscopy. 52・8. 1111-1114 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] T. Shimosaka, T. Kitamori, A. Harata, T. Sawada: "Mulichannel Fluorescence Detector Using Optical Fiber Delay Lines for Variable-Observation-Angle Fluorescence Spectroscopy Under Normal Incidence Conditions" Applied Spectroscopy. 52・2. 308-311 (1998)

    • Related Report
      1998 Annual Research Report

URL: 

Published: 1998-04-01   Modified: 2016-04-21  

Information User Guide FAQ News Terms of Use Attribution of KAKENHI

Powered by NII kakenhi