Single molecule Fluorescence in situ Hybridization (smFISH) for mRNA provides a powerful quantitative handle on expression from endogenous gene loci. While the method has been widely applied in cells in culture, applications to primary tissue samples remain fewer, and often use involved cryosectioning. Even apart from quantitative access to absolute transcript counts in specific tissue volumes, many other advantages of smFISH can be envisaged in tissue samples. Primary among these are the ability to report on subtle differences in expression among different cell types within a tissue, and the ability to correlate the expression from different target genes. Here, we present a modified method of smFISH applicable on various primary wholemount tissues from the fruit fly Drosophila melanogaster, and show the efficacy of the method in a variety of larval and adult tissue, and embryos. We also combine smFISH in tissue with immunofluorescence to demonstrate the possibility of capturing transcriptional and translational aspects of gene expression in the same tissue. Given the widespread use of Drosophila melanogaster as a model system in Developmental Biology and Genetics, such methods are likely to be of wide interest and could yield rich information about gene expression in tissues from this organism.

中文翻译：

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单转录物成像检测全黑果蝇组织中的基因表达

mRNA 的单分子荧光原位杂交 (smFISH) 为内源基因位点的表达提供了强大的定量处理。虽然该方法已广泛应用于培养细胞，但对原代组织样本的应用仍然较少，并且经常使用涉及冷冻切片。即使除了对特定组织体积中的绝对转录计数进行定量访问之外，smFISH 的许多其他优势也可以在组织样本中发挥作用。其中最主要的是能够报告组织内不同细胞类型之间表达的细微差异，以及关联不同靶基因表达的能力。在这里，我们提出了一种改进的 smFISH 方法，适用于来自黑腹果蝇的各种初级整体组织，并展示了该方法在各种幼虫和成虫组织以及胚胎中的功效。我们还将组织中的 smFISH 与免疫荧光相结合，以证明在同一组织中捕获基因表达的转录和翻译方面的可能性。鉴于黑腹果蝇作为发育生物学和遗传学中的模型系统的广泛使用，此类方法可能引起广泛关注，并且可以产生有关该生物组织中基因表达的丰富信息。