Wu Meilin, Lambatan Vanessa, Guo Peng, Joiner William J
Department of Pharmacology, University of California, San Diego, La Jolla, CA 92093, USA.
Department of Cellular and Molecular Medicine, University of California, San Diego, La Jolla, CA 92093, USA; Nikon Imaging Center, University of California, San Diego, La Jolla, CA 92093, USA.
STAR Protoc. 2025 Mar 21;6(1):103615. doi: 10.1016/j.xpro.2025.103615. Epub 2025 Jan 31.
Visualizing the expression of mRNAs using traditional in situ hybridization is often hampered by obstacles including weak signal, high background, and poor probe specificity. Here, we present a protocol utilizing RNAscope (ACD) to overcome these obstacles and detect multiple types of mRNAs simultaneously in whole-mount adult Drosophila brains. We further describe how mRNAs can be reliably quantified in any cells that can be targeted by common binary expression systems such as Gal4/UAS and labeled by immunohistochemistry. For complete details on the use and execution of this protocol, please refer to De et al..
使用传统的原位杂交技术可视化mRNA的表达常常受到诸如信号微弱、背景高和探针特异性差等障碍的阻碍。在这里，我们提出一种利用RNAscope（ACD）的方案来克服这些障碍，并在成年果蝇全脑标本中同时检测多种类型的mRNA。我们进一步描述了如何在任何可被常见二元表达系统（如Gal4/UAS）靶向并通过免疫组织化学标记的细胞中可靠地定量mRNA。有关此方案的使用和执行的完整详细信息，请参考De等人的研究。
