title: High-resolution fluorescent in situ hybridization of Drosophila embryos and tissues.
abstract: INTRODUCTIONFluorescent in situ hybridization (FISH) is commonly used to analyze the three-dimensional distribution of RNAs in intact embryos and tissues. Tyramide signal amplification (TSA) significantly increases the sensitivity and resolution of FISH probe signals. This protocol includes optimized TSA-FISH procedures for Drosophila embryos, ovaries, and larval tissues. Instructions are given for the preparation of RNA probes, the collection and fixation of tissues, and the hybridization and TSA-mediated detection of probes, including options for high-throughput processing in 96-well plates. Variations of the procedure for RNA-RNA and RNA-protein costaining are also described.
authors.affiliation: Donnelly Centre for Cellular and Biomolecular Research, University of Toronto, Toronto, ON, M5S 3E1, Canada.
authors.forename: Eric
authors.initials: E
authors.lastname: Lécuyer
authors.forename: Aleksandar S
authors.initials: AS
authors.lastname: Necakov
authors.forename: Lucía
authors.initials: L
authors.lastname: Cáceres
authors.forename: Henry M
authors.initials: HM
authors.lastname: Krause