Friedrich Miescher Institute for Biomedical Research, Basel, Switzerland.
University of Basel, Basel, Switzerland.
Methods Mol Biol. 2022;2537:173-183. doi: 10.1007/978-1-0716-2521-7_10.
Visualization of single mRNA molecules in fixed cells can be achieved using single molecule fluorescent in situ hybridization (smFISH). This approach enables accurate quantification of mRNA numbers and localization at a single-cell level. To ensure reliable results using smFISH, it is critical to use fluorescent probes that are highly specific to their RNA target. To facilitate probe design, we have created anglerFISH, a user-friendly command-line based pipeline. In this chapter, we present how to perform a smFISH experiment using user-designed and labeled probes.
在固定细胞中可视化单个 mRNA 分子可以使用单分子荧光原位杂交 (smFISH) 来实现。这种方法可以实现 mRNA 数量的精确定量和在单细胞水平上的定位。为了确保使用 smFISH 获得可靠的结果，使用高度特异性针对其 RNA 靶标的荧光探针至关重要。为了便于探针设计，我们创建了anglerFISH，这是一个用户友好的基于命令行的流水线。在本章中，我们介绍如何使用用户设计和标记的探针进行 smFISH 实验。
