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Development of full-automatic in situ hybridization system for Diagnosis

Research Project

Project/Area Number 08557020
Research Category

Grant-in-Aid for Scientific Research (A)

Allocation TypeSingle-year Grants
Section展開研究
Research Field Experimental pathology
Research InstitutionOsaka University

Principal Investigator

NOMURA Shintaro  Osaka University Medical School, Associate professor, 医学部, 助教授 (80159087)

Co-Investigator(Kenkyū-buntansha) HOJOU Shoji  Sakura Instrument, Pathological Research laboratory, Head, 開発本部・病理学研究室, 室長
KITAMURA Yukihiko  Osaka University Medical School, Professor, 医学部, 教授 (70028520)
北条 昭次  (株)サクラ精機, 開発本部病理学研究室, 室長
Project Period (FY) 1996 – 1998
Project Status Completed (Fiscal Year 1998)
Budget Amount *help
¥5,000,000 (Direct Cost: ¥5,000,000)
Fiscal Year 1998: ¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 1997: ¥3,000,000 (Direct Cost: ¥3,000,000)
KeywordsDigoxigeuin / Virus / DNA / mRNA / Pathological Diagnois / in situ hybrildization / Hybridization / Pathological Diagnosis / ウィルス / in situハイブリダイゼーション / ケニルミネッセンス / TSA / ウイルス検出 / 全自動システム / 抗体反応 / 銀増感法 / 全自動検出システム
Research Abstract

We first tested the availability of commercially available "Full automatic histological staining system" and "Full automatic immunostaining system for in situ hybridization system. The combination of both systems did work considarably well in not only to detect viral DNA but mRNAs. However, we recognized that some parts of the instrument need to much more heat resistant for long term use. So we changed some of the parts to the goods resistant to high temperature. Next we investigated the TSA, the amplifier to detect signal. We confirmed the detection limit of the signal raised to approximately 50 times than usual system. This amplifier would be useful for obtaining stable result of in situ hybridization. Finally we tested if we can detect gene amplified product of oncogenes such as ras and sam-1 from sample obtained operation. As a result, we detected amplified sam-1 gene at the efficiency of 30% in the sample. This value was consistent from that obtained by Southern blotting. However, it was difficult to detect amplified gene from pathological samples which were already fixed by formaline. This may due to the regradation of DNA during fixiation. We still need to search optical condition to detect nucleic acid from pathological samples.

Report

(4 results)
  • 1998 Annual Research Report   Final Research Report Summary
  • 1997 Annual Research Report
  • 1996 Annual Research Report
  • Research Products

    (30 results)

All Other

All Publications (30 results)

  • [Publications] M.Sato, S.Nomura, et al.: "Transcriptional regulation of o*leopont*** gene in vivo by P*BP**A/CBFA 1 and T*** 1 in the skeletal tissues"Oncogene. 17. 1517-1525 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] S.Ohshima, S.Nomura, et al.: "IL-6 plays a key role in the development of antigen-induced arthrili*sis"Proc. Natol, Acad. Sci. U.S.A. 95. 8222-8226 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] A.Suzuki, S.Nomura, et al.: "Localization of mRNA for trkB isoforms and p75 in the rat retin*l ganglion cells"J. of Neurosci. Res.. 54. 27-37 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] J.Kosaka, S.Nomura, et al.: "Differential localization and expression of α and * isoenzymes of protein kinase C in the rat retina"J. of Neurosci. Res.. 54. 655-663 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] M.Suoda, S.Nomura, et al.: "Maturational disturbance of chondrocytes in CBfa-1,-deficient **i*o"Dev. Dynamics. (印刷中).

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] K.Terai, S.Nomura, et al.: "The role of osteopo*tin in the bone re**o*leli*y caused by mechanical stress"J. of Bone and Mineral Res.. (印刷中).

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] M. Sato, S. Nomura et al.: "Transcriptional regulation of osteopontin gene in vivo by PEBP2A/CBFA1 and Ets-1 in the skeletal tissues"Oncogene. 17. 1517-1525 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] S. Ohshima, S. Nomura et al.: "IL-6 plays a key role in the development of antigen-induced arthritis (AIA)"Proc. Natl. Acad. Sci.. 95. 8222-8226 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] A. Suzuki, S. Nomura et al.: "Localization of mRNA for TrkB isoforms and p75 in the rat retinal ganglion cells"J. of Neurosci Res.. 54. 27-37 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] J. Kosaka, S. Nomura. Et al.: "Differential localization and expression of A and B isoenzymes of protein kinase C in the rat retina"J. of Neurosci. Res.. 54. 655-663 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] A. Inada, S. Nomura et al.: "Maturational disturbance of chondrocyte in cbfa1-deficient mice"Dev. Dynamics.. (in press).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] K. Terai, S. Nomura, et al.: "The role of osteopontin in the bone remodellijng caused by mechanical stress"J. Bone and Mineral. (in press).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] M.Sato,S.Nomura,et al: "Transcriptional regulation of osteoportor gene in vivo by PFFP2αA/cRFA 1 and Tets1 in the skeletal tissues" Oncogene. 17. 1517-1525 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] S.Ohshima,S.Nomura et al: "Ic-b playsas a key role in the development antigen-induced arthikinacing" Proc.Natl.Acad.Sci.U.S.A. 95. 8222-8226 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] A.Suzuki,S.Nomura et al: "Localization of mRNA for trkB isoforms and p75 inthermal retinal geogliometlle" J.of Neurosci.Res. 54. 27-37 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] T.Kosaka,S.Nomura et al: "Differential localization and expression of α and isoenzymes of protein kinase c in the rat retimes" J.of Neurosci.Res. 54. 655-663 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] M.Onoda,S.Nomura et al: "Maturational disturbance of chometrocytes in CBfa-1-deficient using" Dev.Dynanics. 印刷中.

    • Related Report
      1998 Annual Research Report
  • [Publications] K.Terai,S.Nomura et al: "The role of osteopositin in the Kinase sensedeling caused by mechanical stress" T.of Bone and Mineral Res. 印刷中.

    • Related Report
      1998 Annual Research Report
  • [Publications] T.Komori: "Targeted disruption of CBFAI/BERPαA gene results in complete lackol bone formation due to the maturational arrest of asteoblerls" Cell. 89. 755-764 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] S.Nomura: "Cellular localization of thrombopoietin mRNA by in situ lybridization" Exp.Haemutol.25. 565-572 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] N.Yasui: "Distraction ostepgenesis in a rat model three different models of ossification" J.of Bone and Joint Surgery. 79. 824-830 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] M.Sugimoto: "Impared expression of non-collugenous bone matrix protein in DNA during fracture bealing in ascorbic-acid deficient rats" J.Bone and Mineral Res.13. 271-277 (1998)

    • Related Report
      1997 Annual Research Report
  • [Publications] K.Fukuo: "Possible participation of Fas mediated apoptosis in the mechanism of atleroscrelesis" Gastoenleology. 43. 34-42 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] N.Chiba: "Differentiation dependent expression and distinct subecllular localization of the Protonogene product PEBP 21/CBFP in muscle development" Oncogene. 14. 2543-2552 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] K.Takebayashi: "Reseesire phenotype disployed by dominant negative MITF mutant is a result of impared nuclear localization potential" Hol.Cell.Biol.16. 1203-1211 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] T.Kimura: "Expression of oxytocin receptor in human pregnant myometrium" Endocrinology. 137. 780-785 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Y.Nakajima: "Identification of a cell types of adontoma-life clustere in microphthalmic (mi/mi) mice using in situ hybridization" Pathol.Int.46. 743-750 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] K.Tokunaga: "Human osteosarcoma(OST)induces mouse reactive bone formation in yenograft system" Bone. 19. 447-454 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] S.Nomura: "Cellular localization of thrombopoietin mRNA in the liver by in situ hybridization" Exp Haematol. (印刷中).

    • Related Report
      1996 Annual Research Report
  • [Publications] T.Takemura: "Localization of bone morplwgenetic protein-4 messenger RNA in developing mouse cochlea" Hearing Res.95. 26-32 (1996)

    • Related Report
      1996 Annual Research Report

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

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