ArticleMethods in molecular biology (Clifton, N.J.)2026
smFISH: Quantifying and Mapping RNA at the Single-Molecule Level.
Article in Methods in molecular biology (Clifton, N.J.), 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.
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Abstract
RNA localization regulates spatial and temporal control of protein synthesis, influencing processes such as development, neuronal signaling, cell migration, and stress responses. Single-molecule fluorescence in situ hybridization (smFISH) enables direct visualization and quantification of individual transcripts within cells, providing both absolute counts and subcellular localization of RNAs. Here, we present a detailed, step-by-step protocol for smFISH in adherent mammalian cells, including probe design and labeling, sample preparation, hybridization, imaging, and analysis. This guide offers a reproducible workflow that facilitates the study of RNA localization and expression at single-molecule resolution.
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