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Isolation of Tissue Macrophages in Adult Zebrafish

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Tissue-Resident Macrophages

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2713))

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Abstract

Tissue macrophages are essential components of the immune system that also play key roles in vertebrate development and homeostasis, including in zebrafish, which has gained popularity over the years as a translational model for human disease. Commonly, zebrafish macrophages are identified based on expression of fluorescent transgenic reporters, allowing for real-time imaging in living animals. Several of these lines have also proven instrumental to isolate pure populations of macrophages in the developing embryo and larvae using fluorescence-activated cell sorting (FACS). However, the identification of tissue macrophages in adult fish is not as clear, and robust protocols are needed that would take into account changes in reporter specificity as well as the heterogeneity of mononuclear phagocytes as fish reach adulthood. In this chapter, we describe the methodology for analyzing macrophages in various tissues in the adult zebrafish by flow cytometry. Coupled with FACS, these protocols further allow for the prospective isolation of enriched populations of tissue-specific mononuclear phagocytes that can be used in downstream transcriptomic and/or epigenomic analyses. Overall, we aim at providing a guide for the zebrafish community based on our expertise investigating the adult mononuclear phagocyte system.

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Authors and Affiliations

  1. Institut de Recherche Interdisciplinaire en Biologie Humaine et Moléculaire (IRIBHM), Brussels, Belgium

    Mireia Rovira, Jennifer Pozo, Magali Miserocchi & Valérie Wittamer

  2. ULB Neuroscience Institute (UNI), Université Libre de Bruxelles (ULB), Brussels, Belgium

    Mireia Rovira, Jennifer Pozo, Magali Miserocchi & Valérie Wittamer

Authors
  1. Mireia Rovira
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  2. Jennifer Pozo
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  3. Magali Miserocchi
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  4. Valérie Wittamer
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Corresponding author

Correspondence to Valérie Wittamer .

Editor information

Editors and Affiliations

  1. Life & Medical Sciences Institute (LIMES), University of Bonn, Bonn, Germany

    Elvira Mass

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© 2024 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

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Rovira, M., Pozo, J., Miserocchi, M., Wittamer, V. (2024). Isolation of Tissue Macrophages in Adult Zebrafish. In: Mass, E. (eds) Tissue-Resident Macrophages. Methods in Molecular Biology, vol 2713. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3437-0_5

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  • DOI: https://doi.org/10.1007/978-1-0716-3437-0_5

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-3436-3

  • Online ISBN: 978-1-0716-3437-0

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