mRNA analysis of single living cells.
mRNA analysis of single living cells.
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DOI：
10.1186/1477-3155-1-2
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发表时间：
2003-02-14
影响因子：
10.2
通讯作者：
Ikai, Atsushi
中科院分区：
文献类型：
--
作者：
Osada, Toshiya;Uehara, Hironori;Ikai, Atsushi
单个活细胞中特定基因表达的分析可能成为细胞生物学的一项重要技术。到目前为止，还没有一种方法可以在不杀死或破坏活细胞的情况下检测活细胞中的 mRNA。我们在这里开发了一种使用原子力显微镜（AFM）检查活细胞基因表达的新方法。将 AFM 尖端插入活细胞中以提取 mRNA。使用RT-PCR、巢式PCR和定量PCR对获得的mRNA进行分析。这种方法使我们能够检查单个活细胞的时间依赖性基因表达，而不会对细胞造成严重损害。
Analysis of specific gene expression in single living cells may become an important technique for cell biology. So far, no method has been available to detect mRNA in living cells without killing or destroying them. We have developed here a novel method to examine gene expression of living cells using an atomic force microscope (AFM). AFM tip was inserted into living cells to extract mRNAs. The obtained mRNAs were analyzed with RT-PCR, nested PCR, and quantitative PCR. This method enabled us to examine time-dependent gene expression of single living cells without serious damage to the cells.