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2005 Fiscal Year Final Research Report Summary

Development of high-throughput analytical system for intracellular molecules with quantum dots

Research Project

Project/Area Number 16390107
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Human pathology
Research InstitutionYamaguchi University

Principal Investigator

SASAKI Kohsuke  Yamaguchi University, School of Medicine, Professor, 医学部, 教授 (80116722)

Co-Investigator(Kenkyū-buntansha) KAWAUCHI Shigeto  Yamaguchi University, School of Medicine, Assistant Professor, 医学部, 講師 (80284511)
OGA Atunori  Yamaguchi University, School of Medicine, Research Associate, 医学部, 助手 (90243633)
Project Period (FY) 2004 – 2005
KeywordsQuantum Dots / Cytomics / Immunocytochemisty / Cytometry
Research Abstract

Fluorescent semiconductor quantum dots (QDs) are a novel class of fluorescent probe with optical properties that differ from those of conventional fluorochromes. QDs emit bright fluorescence at visible wavelengths, depending on size of the particle, and in addition this bright fluorescence is resistant to photobleaching. In this project, we aimed to demonstrate that QDs are useful to quantify intracellular materials and applied to cytometry. First, QDs were substituted for organic fluorochromes in immunohistochemistry. Some intracellular/nuclear antigens were detected by secondary antibodies labeled with QDs, but some antigens were not. This may be caused by the difference of physiochemical property between two types of fluoroprobes and we tried various fixations and dying procedure to find optimal condition for immunohistochemistry with QDs. For example, samples needed treatment with Triton X-100 and microwave exposure in sodium citrate buffer (pH6) for the purpose of the detection of … More intranuclear proliferating cell nuclear antigen (PCNA). We plan to determine the optimal methods for various antigens. We verified whether QDs bind to their targets stoichiometrically as well as classical fluorochromes. We labeled PCNA in nuclei of PC-14 cells immunocytochemically with QDs and Alexa 488 concomitantly and measured the level of intranuclear PCNA expression during cell cycle with laser scanning cytometry. At the same time, nuclear DNA stained with propidium iodide (PI) was measured. The bivariate DNA/PCNA content cytograms for QDs and Alexa 488 fluorescence were dome-shaped and very similar to each other. These indicated that immunocytochemistry with QDs under optimal condition provided the data equivalent to those with organic fluorescence dyes and QDs could be useful fluorochromes in cytometry. However, we didn't always succeed to detect fluorescence intensity by LSC or flow cytometer and we need further examination for immunocytochemical methods and optical condition in cytometry. Less

  • Research Products

    (18 results)

All 2006 2005 2004

All Journal Article (18 results)

  • [Journal Article] Biological characteristics in bladder cancer depend on the type of genetic instability.2006

    • Author(s)
      Yamamoto Y
    • Journal Title

      Clin Cancer Res. 12

      Pages: 2752-2758

    • Description
      「研究成果報告書概要(和文)」より
  • [Journal Article] Relation of DNA ploidy to gentic aberrations detected by chromosomal CGH and FISH in gastric adecnoacinomas2006

    • Author(s)
      Furuya T
    • Journal Title

      Oncol Rep. 15

      Pages: 1491-1496

    • Description
      「研究成果報告書概要(和文)」より
  • [Journal Article] Molucular cytogenetic analysis of oral squamous cell carcinomas by comparative genomic hybridization, spectral karyotyping, and fluorescence in situ hybridization.2006

    • Author(s)
      Uchida T
    • Journal Title

      Cancer Genet Cytogenet. 167

      Pages: 109-116

    • Description
      「研究成果報告書概要(和文)」より
  • [Journal Article] Biological characteristics in bladder cancer depend on the type of genetic instability2006

    • Author(s)
      Yamamoto Y.
    • Journal Title

      Clin Cancer Res. 12

      Pages: 2752-2758

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] Relation of DNA ploidy to genetic aberrations detected by chromosomal CGH and FISH in gastric adenocarcinomas2006

    • Author(s)
      Furuya T.
    • Journal Title

      Oncol Rep. 15

      Pages: 1491-1496

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] Molecular cytogenetic analysis of oral squamous cell carcinomas by comparative genomic hybridization, spectral karyotyping, and fluorescence in situ hybridization.2006

    • Author(s)
      Uchida T.
    • Journal Title

      Cancer Genet Cytogenet. 167

      Pages: 109-116

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] Large cell neuroendocrine carcinoma of the uterine cervix with cytogenetic analysis by comparative genomic hybridization : acase syudy.2005

    • Author(s)
      Kawauchi S.
    • Journal Title

      Hum Pathol. 36

      Pages: 1096-1100

    • Description
      「研究成果報告書概要(和文)」より
  • [Journal Article] Large cell nueroendocrine carcinoma of the uterine cervix with cytogenetic anlysis by comparative genomic hybridization : a case study.2005

    • Author(s)
      Kawauchi S.
    • Journal Title

      Hum Pathol. 36

      Pages: 1096-1100

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] analysis of DNA copy number aberrations in hepatitis C virus-associated hepatocellular carcinomas by conventional CGH and array CGH2004

    • Author(s)
      Hashimoto K
    • Journal Title

      Mod Pathol 17

      Pages: 617-622

    • Description
      「研究成果報告書概要(和文)」より
  • [Journal Article] multicolor FISH and Cytometric analyses allow classification of urothelial carcinomas into two subtypes, low- and high-grade tumors.2004

    • Author(s)
      Ikemoto K
    • Journal Title

      Int J Oncol. 25

      Pages: 893-898

    • Description
      「研究成果報告書概要(和文)」より
  • [Journal Article] Centrosome hyperamplification predicts progression and tumor reccurence in baldder cancer.2004

    • Author(s)
      Yamamoto Y
    • Journal Title

      Clin Cancer Res. 10

      Pages: 6449-6455

    • Description
      「研究成果報告書概要(和文)」より
  • [Journal Article] Significance of beta-catenin and pRB pathway components in malignant ovarian germ cell tumors : INK4A promotor CpG island methylation is associated with cell proliferation.2004

    • Author(s)
      Kawauchi S.
    • Journal Title

      J Pathol. 204

      Pages: 268-276

    • Description
      「研究成果報告書概要(和文)」より
  • [Journal Article] Comparative genomic hybridization analysis for pancreatic cancer specimens obtained by endoscopic ultrasonography-guided fine-needle aspiration.2004

    • Author(s)
      Kitoh H
    • Journal Title

      J Gastroenterol. 40

      Pages: 511-517

    • Description
      「研究成果報告書概要(和文)」より
  • [Journal Article] Analysis of DNA copy number aberrations in hepatitis C virus-associated hepatocellular carcinomas by conventional CGH and array CGH2004

    • Author(s)
      Hashimoto K
    • Journal Title

      Mod Pathol. 17

      Pages: 617-622

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] Multicolor FISH and Cytometric analyses allow classification of urothelial carcinomas into two subtypes, low- and high-grade tumors.2004

    • Author(s)
      Ikemoto K.
    • Journal Title

      Int J Oncol. 25

      Pages: 893-898

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] Centrosome hyperamplification predicts progression and tumor recurrence in bladder cancer.2004

    • Author(s)
      Yamamoto Y.
    • Journal Title

      Clin cancer Res. 10

      Pages: 6449-6455

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] Significance of beta-catenin and pPB pathway components in malignant ovarian germ cell tumor : INK4A promoter CpG island methylation is associated with cell proliferation.2004

    • Author(s)
      Kawauchi S
    • Journal Title

      J Pathol. 204

      Pages: 268-276

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] Comparative genomic hybridization analysis for pancreatic cancer specimens obtained by endoscopic ultrasonography-guided fine-needle aspiration.2004

    • Author(s)
      Kitoh H.
    • Journal Title

      J Gastroenterol. 40

      Pages: 511-517

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

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Published: 2007-12-13  

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