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One FISH, dFISH, Three FISH: Sensitive Methods of Whole-Mount Fluorescent In Situ Hybridization in Freshwater Planarians

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In Situ Hybridization Methods

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

Abstract

As freshwater planarians (flatworms) are further developed as a model system, the most valuable tool continues to be in situ hybridization (ISH) analysis of gene expression in whole mount adult animals (WISH). Multiple hurdles have been overcome during the optimization of a standard protocol for colorimetric, single color detections. However, gene function studies on planarians have evolved to the point where virtually all investigations require the analysis of multiple RNAs, simultaneously. Thus, considerable effort by the entire planarian community has been put forth to create adequate methodologies towards this goal. Here we summarize the field, methodological evolution, and describe a new method for more rapid and more sensitive detection of multiple RNAs by multi-fluorescent ISH.

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Acknowledgements

We thank the entire Pearson lab for continuing troubleshooting and improvements on this method. DDRB was funded by the Canadian Institute for Health Research (CIHR) Frederick Banting and Charles Best Canada Graduate Scholarship #201101GSD-277528. BJP was funded by Ontario Institute for Cancer Research (OICR) award #IA-026.

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Authors and Affiliations

  1. Program in Developmental and Stem Cell Biology, The Hospital for Sick Children, Toronto, ON, Canada

    David D. R. Brown & Bret J. Pearson

  2. Department of Molecular Genetics, University of Toronto, Toronto, ON, Canada

    David D. R. Brown & Bret J. Pearson

  3. MaRS Centre, Ontario Institute for Cancer Research, 101 College St., Room 11-704, Toronto, ON, Canada, M5G1L7

    Bret J. Pearson

Authors
  1. David D. R. Brown
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  2. Bret J. Pearson
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Correspondence to Bret J. Pearson .

Editor information

Editors and Affiliations

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

    Giselbert Hauptmann

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Brown, D.D.R., Pearson, B.J. (2015). One FISH, dFISH, Three FISH: Sensitive Methods of Whole-Mount Fluorescent In Situ Hybridization in Freshwater Planarians. 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_7

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  • DOI: https://doi.org/10.1007/978-1-4939-2303-8_7

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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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