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. 2022:2537:173-183.
doi: 10.1007/978-1-0716-2521-7_10.

Design, Labeling, and Application of Probes for RNA smFISH

Ewa Piskadlo  1 , Bastian Th Eichenberger  1   2 , Luca Giorgetti  1 , Jeffrey A Chao  3
Affiliations

Design, Labeling, and Application of Probes for RNA smFISH

Ewa Piskadlo et al. Methods Mol Biol. 2022.

Abstract

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.

Keywords: Command line interface (CLI); FISH probe design; Fluorescence labeling of oligonucleotides; Nextflow workflow; Single-molecule fluorescence in situ hybridization (smFISH).

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