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Multicolored Visualization of Transcript Distributions in Drosophila Embryos

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In Situ Hybridization Methods

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

Abstract

Despite large-scale gene expression profiling studies, it is still often required to precisely characterize the localization of different transcripts in relation to each other to determine unique and overlapping expression sites at a cellular level. We describe here a versatile protocol for simultaneous examination of three unique mRNA expression patterns in Drosophila melanogaster embryos. Three differently labeled antisense RNA probes are hybridized together to the embryos and detected by sequential alkaline phosphatase-based immunohistochemistry. Transcript distributions are revealed by colorimetric enzymatic reactions that permit to highlight each mRNA expression pattern by a differing and contrasting cellular color precipitate. We provide tips and tricks for each critical step helpful for successful application of the tricolor whole-mount in situ hybridization (WISH) method.

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Acknowledgements

We thank Prof. Dr. Ulrich Theopold and the Wenner Gren Institute of Stockholm University for their support, while preparing this manuscript. This method was initially developed during G.H.’s stay in the laboratory of Dr. Thomas Gerster at the Biozentrum of the Universität Basel.

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Authors and Affiliations

  1. Department of Molecular Biosciences, The Wenner-Gren Institute, Stockholm University, Svante Arrhenius väg 20C, 10691, Stockholm, Sweden

    Iris Söll & Giselbert Hauptmann

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  1. Iris Söll
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  2. Giselbert Hauptmann
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Correspondence to Giselbert Hauptmann .

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Editors and Affiliations

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

    Giselbert Hauptmann

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Söll, I., Hauptmann, G. (2015). Multicolored Visualization of Transcript Distributions in Drosophila Embryos. 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_3

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  • DOI: https://doi.org/10.1007/978-1-4939-2303-8_3

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-2302-1

  • Online ISBN: 978-1-4939-2303-8

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