Department of Physics, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.
Nat Methods. 2011 Apr;8(4 Suppl):S12-9. doi: 10.1038/nmeth.1573. Epub 2011 Mar 30.
High-throughput gene expression screens provide a quantitative picture of the average expression signature of biological samples. However, the analysis of spatial gene expression patterns with single-cell resolution requires quantitative in situ measurement techniques. Here we describe recent technological advances in RNA fluorescence in situ hybridization (FISH) techniques that facilitate detection of individual fluorescently labeled mRNA molecules of practically any endogenous gene. These methods, which are based on advances in probe design, imaging technology and image processing, enable the absolute measurement of transcript abundance in individual cells with single-molecule resolution.
高通量基因表达筛选提供了生物样本平均表达特征的定量图像。然而，具有单细胞分辨率的空间基因表达模式分析需要定量的原位测量技术。在这里，我们描述了 RNA 荧光原位杂交 (FISH) 技术的最新技术进展，这些技术有助于检测几乎任何内源性基因的单个荧光标记的 mRNA 分子。这些方法基于探针设计、成像技术和图像处理方面的进展，能够以单分子分辨率对单个细胞中的转录物丰度进行绝对测量。
