Abstract：
The RNA fluorescence in situ hybridization (FISH) technique combined with immunostaining is a powerful method to visualize a specific transcript and a protein of interest simultaneously. Although whole-mount RNA FISH is routinely used to determine RNA intracellular localization, a detailed picture of RNA distribution in complex tissues remains a challenge. The main problem is the various permeability of morphologically different cells within a tissue. We overcome this challenge by developing an approach based on differential permeabilization treatment of tissue specimens. We have tested and optimized conditions for RNA FISH combined with immunofluorescent staining (RNA FISH/IF) to detect the maternal telomeric retrotransposon HeT-A RNPs in the Drosophila ovaries and syncytial embryos. Methods described here are applicable to a broad variety of biological tissue specimens.
摘要：
结合免疫染色的RNA荧光原位杂交(FISH)技术是同时可视化特定转录物和感兴趣的蛋白质的强大方法。尽管完整RNAFISH通常用于确定RNA细胞内定位，复杂组织中RNA分布的详细图片仍然是一个挑战。主要问题是组织内形态上不同的细胞的各种渗透性。我们通过开发一种基于组织标本差异渗透处理的方法来克服这一挑战。我们已经测试和优化了RNAFISH结合免疫荧光染色（RNAFISH/IF）的条件，以检测果蝇卵巢和合胞胚胎中的母体端粒逆转录转座子HeT-ARNP。这里描述的方法适用于多种生物组织标本。
