ArticleMethods in molecular biology (Clifton, N.J.)2026
Single-Molecule RNA Fluorescence In Situ Hybridization (smRNA-FISH) Labeling and Imaging of mRNA to Determine Subcellular Localization.
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
The protocol reported in this chapter describes methodologies for labeling of RNA by single-molecule RNA fluorescence in situ hybridization (smRNA-FISH), performing imaging confocal microscopy to image RNAs, and post-acquisition analyses. Specifically, we describe methods to design and generate smRNA-FISH probes, label RNAs in situ, and image RNAs. We also describe stepwise post-acquisition analyses using Image J to determine the absolute quantity, subcellular localization, and translation status of messenger RNAs. We provide examples of labeling full-length RNAs and their subspecies, quantifying RNA localization to organelles and ribonucleoprotein granules, and analyses of the translation status of individual mRNAs.
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