Visualization of single RNA transcripts in situ

Andrea M. Femino, Fredric S. Fay, Kevin Fogarty, Robert H. Singer

Research output: Contribution to journalArticlepeer-review

743 Scopus citations

Abstract

Fluorescence in situ hybridization (FISH) and digital imaging microscopy were modified to allow detection of single RNA molecules. Oligodeoxynucleotide probes were synthesized with five fluorochromes per molecule, and the light emitted by a single probe was calibrated. Points of light in exhaustively deconvolved images of hybridized cells gave fluorescent intensities and distances between probes consistent with single messenger RNA molecules. Analysis of β-actin transcription sites after serum induction revealed synchronous and cyclical transcription from single genes. The rates of transcription initiation and termination and messenger RNA processing could be determined by positioning probes along the transcription unit. This approach extends the power of FISH to yield quantitative molecular information on a single cell.

Original languageEnglish (US)
Pages (from-to)585-590
Number of pages6
JournalScience
Volume280
Issue number5363
DOIs
StatePublished - Apr 24 1998

ASJC Scopus subject areas

  • General

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