title: Simultaneous multicolor detection of RNA and proteins using super-resolution microscopy.
abstract: A number of non-membranous cellular bodies have been identified in higher eukaryotes, and these bodies contain a specific set of proteins and RNAs that are used to fulfill their functions. The size of these RNA-containing cellular bodies is usually on a submicron scale, making it difficult to observe fine structures using optical microscopy due to the diffraction limitation of visible light. Recently, microscope companies have released super-resolution microscopes that were developed using different principles, enabling the observation of sub-micron structures not resolvable in conventional fluorescent microscopy. Here, we describe multi-color fluorescent in situ hybridization techniques optimized for the simultaneous detection of RNA and proteins using super-resolution microscopy, namely structured illumination microscopy (SIM).
authors.affiliation: RNA Biology Laboratory, RIKEN, 2-1 Hirosawa, Wako 351-0198, Japan.
authors.forename: Mari
authors.initials: M
authors.lastname: Mito
authors.affiliation: Institute for Genetic Medicine, Hokkaido University, Sapporo 060-0815, Japan.
authors.forename: Tetsuya
authors.initials: T
authors.lastname: Kawaguchi
authors.affiliation: Institute for Genetic Medicine, Hokkaido University, Sapporo 060-0815, Japan.
authors.forename: Tetsuro
authors.initials: T
authors.lastname: Hirose
authors.affiliation: RNA Biology Laboratory, RIKEN, 2-1 Hirosawa, Wako 351-0198, Japan. Electronic address: nakagawas@riken.jp.
authors.forename: Shinichi
authors.initials: S
authors.lastname: Nakagawa
mesh_terms: D000906:Antibodies / Q000737:chemistry; D001710:Biotin / Q000737:chemistry; D002460:Cell Line; D004265:DNA Helicases / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D004268:DNA-Binding Proteins; D004072:Digitonin / Q000737:chemistry; D016650:Fluorescein-5-isothiocyanate / Q000737:chemistry; D006241:Haptens / Q000737:chemistry; D006801:Humans; D021621:Imaging, Three-Dimensional; D017404:In Situ Hybridization, Fluorescence / Q000379:methods*; D008856:Microscopy, Fluorescence / Q000379:methods*; D034521:Nuclear Matrix-Associated Proteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D009687:Nuclear Proteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D050810:Octamer Transcription Factors / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D000072670:PTB-Associated Splicing Factor / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D015347:RNA Probes / Q000737:chemistry*; D062085:RNA, Long Noncoding / Q000032:analysis* / Q000235:genetics / Q000378:metabolism; D034702:RNA-Binding Protein FUS / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D013194:Staining and Labeling / Q000379:methods; D014157:Transcription Factors / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D014158:Transcription, Genetic
chemical_list: D000906:Antibodies; D004268:DNA-Binding Proteins; C000599550:FUS protein, human; D006241:Haptens; C577276:NEAT1 long non-coding RNA, human; C083214:NONO protein, human; D034521:Nuclear Matrix-Associated Proteins; D009687:Nuclear Proteins; D050810:Octamer Transcription Factors; D000072670:PTB-Associated Splicing Factor; D015347:RNA Probes; D062085:RNA, Long Noncoding; D034702:RNA-Binding Protein FUS; D016601:RNA-Binding Proteins; D014157:Transcription Factors; D001710:Biotin; C084108:SMARCA4 protein, human; D004265:DNA Helicases; D016650:Fluorescein-5-isothiocyanate; D004072:Digitonin
keywords: Cy2; In situ hybridization; Structured illumination microscopy (SIM); Super-resolution microscopy; TDE