title: Quantitative gene expression analysis in Caenorhabditis elegans using single molecule RNA FISH.
abstract: Advances in fluorescent probe design and synthesis have allowed the uniform in situ labeling of individual RNA molecules. In a technique referred to as single molecule RNA FISH (smRNA FISH), the labeled RNA molecules can be imaged as diffraction-limited spots and counted using image analysis algorithms. Single RNA counting has provided valuable insights into the process of gene regulation. This microscopy-based method has often revealed a high cell-to-cell variability in expression levels, which has in turn led to a growing interest in investigating the biological significance of gene expression noise. Here we describe the application of the smRNA FISH technique to samples of Caenorhabditis elegans, a well-characterized model organism.
authors.affiliation: Institute of Cellular Biology and Pathology, First Faculty of Medicine, Charles University in Prague, Albertov 4, 128 00 Prague, Czech Republic. Electronic address: jitka.simandlova@lf1.cuni.cz.
authors.forename: Jitka
authors.initials: J
authors.lastname: Bolková
authors.affiliation: Institute of Cellular Biology and Pathology, First Faculty of Medicine, Charles University in Prague, Albertov 4, 128 00 Prague, Czech Republic. Electronic address: clanctot9@hotmail.com.
authors.forename: Christian
authors.initials: C
authors.lastname: Lanctôt
mesh_terms: D000465:Algorithms; D000818:Animals; D017173:Caenorhabditis elegans / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D004625:Embryo, Nonmammalian / Q000378:metabolism / Q000648:ultrastructure*; D005456:Fluorescent Dyes / Q000737:chemistry; D018507:Gene Expression Regulation, Developmental; D017404:In Situ Hybridization, Fluorescence / Q000379:methods*; D012333:RNA, Messenger / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D059629:Signal-To-Noise Ratio; D000072760:Single Molecule Imaging / Q000379:methods*; D016707:Tissue Fixation / Q000379:methods; D014158:Transcription, Genetic
chemical_list: D005456:Fluorescent Dyes; D012333:RNA, Messenger
keywords: C. elegans; Gene expression; Image analysis; Nascent transcription; RNA FISH; Single molecule analysis