Single-molecule fluorescence in situ hybridization: Quantitative imaging of single RNA molecules

Research output: Contribution to journalShort survey

43 Scopus citations

Abstract

In situ detection of RNAs is becoming increasingly important for analysis of gene expression within and between intact cells in tissues. International genomics efforts are now cataloging patterns of RNA transcription that play roles in cell function, differentiation, and disease formation, and they are demonstrating the importance of coding and noncoding RNA transcripts in these processes. However, these techniques typically provide ensemble averages of transcription across many cells. In situ hybridization-based analysis methods complement these studies by providing information about how expression levels change between cells within normal and diseased tissues, and they provide information about the localization of transcripts within cells, which is important in understanding mechanisms of gene regulation. Multi-color, single-molecule fluorescence in situ hybridization (smFISH) is particularly useful since it enables analysis of several different transcripts simultaneously. Combining smFISH with immunofluorescent protein detection provides additional information about the association between transcription level, cellular localization, and protein expression in individual cells.

Original languageEnglish (US)
Pages (from-to)65-72
Number of pages8
JournalBMB Reports
Volume46
Issue number2
DOIs
StatePublished - Feb 1 2013

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Keywords

  • Breast cancer
  • ERBB2
  • FISH
  • RNA
  • Ribonucleoprotein (RNP)
  • Single-molecule imaging

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

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