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Subcellular Transcript Localization in Drosophila Embryos and Tissues Visualized by Multiplex-FISH

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In Situ Hybridization Methods

Part of the book series: Neuromethods ((NM,volume 99))

Abstract

Determining the spatiotemporal expression dynamics of a gene, or the subcellular localization properties of its encoded RNA, is often a key first step toward elucidating its function. Fluorescent in situ hybridization (FISH) represents the gold standard method for visualizing RNA expression and subcellular localization features in distinct cells, tissue specimens, and whole-mount organisms. This chapter describes a high-resolution FISH protocol for the detection of coding or noncoding RNA expression and localization dynamics in embryos and tissues of the fruit fly, Drosophila melanogaster. Variations of the protocol are proposed for the co-detection of different RNAs and for the co-labeling of RNAs and proteins. The protocol also outlines details for conducting FISH in 96-well plate format, which significantly enhances the throughput and versatility of the procedure.

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Authors and Affiliations

  1. Institut de Recherches Cliniques de Montréal (IRCM), Montréal, QC, Canada

    Julie Bergalet, Carole Iampietro, Xuan-Tam Nguyen, Sulin Oré-Rodriguez, Neal Cody & Eric Lécuyer

  2. Département de Biochimie, Université de Montréal, Montréal, QC, Canada

    Xuan-Tam Nguyen, Sulin Oré-Rodriguez & Eric Lécuyer

  3. Division of Experimental Medicine, McGill University, Montréal, QC, Canada

    Eric Lécuyer

  4. IRCM, RNA Biology Laboratory, 110 Avenue des Pins, Ouest, Montréal, QC, Canada, H2W 1R7

    Eric Lécuyer

  5. Division of Experimental Medicine, McGill University, Institut de Recherches Cliniques de Montréal (IRCM), Montréal, QC, Canada

    Ashley Chin

Authors
  1. Julie Bergalet
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  2. Carole Iampietro
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  3. Ashley Chin
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  4. Xuan-Tam Nguyen
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  5. Sulin Oré-Rodriguez
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  6. Neal Cody
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  7. Eric Lécuyer
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Corresponding author

Correspondence to Eric Lécuyer .

Editor information

Editors and Affiliations

  1. Department of Molecular Biosciences, The Wenner-Gren Institute, Stockholm University, Stockholm, Sweden

    Giselbert Hauptmann

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Bergalet, J. et al. (2015). Subcellular Transcript Localization in Drosophila Embryos and Tissues Visualized by Multiplex-FISH. In: Hauptmann, G. (eds) In Situ Hybridization Methods. Neuromethods, vol 99. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2303-8_19

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  • DOI: https://doi.org/10.1007/978-1-4939-2303-8_19

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-2302-1

  • Online ISBN: 978-1-4939-2303-8

  • eBook Packages: Springer Protocols

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