首页> 中文期刊> 《中国病理生理杂志》 >联合应用实时量RT-PCR和激光显微切割检测单个肝细胞RNA的表达

联合应用实时量RT-PCR和激光显微切割检测单个肝细胞RNA的表达

         

摘要

AIM: The manner in which a cell responds to and influences its environment is ultimately determined by the genes that are expressed. To better understand cellular functions, the isolation of single cells and subsequent quantification of the expressed genes is essential. METHODS: Normal liver tissue was obtained from operation, snapfrozen in liquid nitrogen and sectioned in crystat. Individual hepatocytes were microdissected. RNA was extracted, then reverse transcribed and amplified using real- time quantitative polymerase chain reaction (PCR). RESULTS: Single hepatocytes were dissected by laser beam and catapulted to the microcentrifuge cap which was put above the slide. In this way,cells were collected, RNA was extracted, reverse transcribed to cDNA and used for analysis of RNA expression by realtime quantitative PCR. The amplification results showed that quantitation of the RNA inside the cell was compatible with the number of cells. CONCLUSION: The expression of RNA in single cells can be quantitated successfully by using laser microdissection and real -time PCR. These techniques provide an opportunity to monitor in vivo gene expression levels in single hepatocytes.

著录项

  • 来源
    《中国病理生理杂志》 |2007年第11期|2285-2288|共4页
  • 作者

  • 作者单位

    Department of General Surgery, University of Heidelberg, Im Neuenheimer Feld 110,D - 69120 Heidelberg, Germany;

    Department of General Surgery, Zhongda Hospital, Southeast University, Nanjing 210009. China;

    Department of General Surgery, University of Heidelberg, Im Neuenheimer Feld 110,D - 69120 Heidelberg, Germany;

    Department of General Surgery, University of Heidelberg, Im Neuenheimer Feld 110,D - 69120 Heidelberg, Germany;

    Department of General Surgery, University of Heidelberg, Im Neuenheimer Feld 110,D - 69120 Heidelberg, Germany;

    Department of General Surgery, Zhongda Hospital, Southeast University, Nanjing 210009. China;

    Institute of Pathology, University of Bern, Inselspital , CH -3010 Bern, Switzerland;

    Department of General Surgery, University of Heidelberg, Im Neuenheimer Feld 110,D - 69120 Heidelberg, Germany;

    Department of General Surgery, University of Heidelberg, Im Neuenheimer Feld 110,D - 69120 Heidelberg, Germany;

  • 原文格式 PDF
  • 正文语种 chi
  • 中图分类 病理生理学;
  • 关键词

    Laser microdissction; Real - time quantitative PCR; RNA; Hepatocytes;

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