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Time-Lapse Imaging and Morphometric Analysis of Tracheal Development in Drosophila

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Cell Migration in Three Dimensions

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2608))

Abstract

Detailed and quantitative analyses of the cellular events underlying the formation of specific organs or tissues is essential to understand the general mechanisms of morphogenesis and pattern formation. Observation of live tissues or whole-mount fixed specimens has emerged as the method of choice for identifying and quantifying specific cellular and tissular structures within the organism. In both cases, cell and subcellular structure identification and good quality image acquisition for these analyses are essential. Many markers for live imaging and fixed tissue are now available for detecting cell membranes, subcellular structures, and extracellular structures like the extracellular matrix (ECM). Combination of live imaging and analysis of fixed tissue is ideal to obtain a general and detailed picture of the events underlying embryonic development. By applying morphometric methods to both approaches, we can, in addition, obtain a quantitative evaluation of the specific parameters under investigation in morphogenetic and cell biological studies. In this chapter, we focus on the development of the tracheal system of Drosophila melanogaster, which provides an ideal paradigm to understand the formation of branched tubular organs. We describe the most used methods of imaging and morphometric analysis in tubulogenesis using mainly (but not exclusively) examples from embryonic development. We cover embryo preparation for fixed and live analysis of tubulogenesis, together with methods to visualize larval tracheal terminal cell branching and lumen formation. Finally, we describe morphometric analysis and quantification methods using fluorescent images of tracheal cells.

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Author information

Authors and Affiliations

  1. Department of Genetics, Microbiology and Statistics, School of Biology, University of Barcelona (UB), Barcelona, Spain

    Sofia J. Araújo

  2. Institute of Biomedicine, University of Barcelona (IBUB), Barcelona, Spain

    Sofia J. Araújo

  3. Institute of Molecular Biology of Barcelona (IBMB), CSIC, Parc Científic de Barcelona, Barcelona, Spain

    Marta Llimargas

Authors
  1. Sofia J. Araújo
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  2. Marta Llimargas
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Corresponding authors

Correspondence to Sofia J. Araújo or Marta Llimargas .

Editor information

Editors and Affiliations

  1. Department of Medical Oncology, Cancer Center Amsterdam, Amsterdam University Medical Center, Amsterdam, The Netherlands

    Coert Margadant

1 Electronic Supplementary Material

Movie 1

Tracheal time-lapse imaging. In vivo tracheal cell visualization during branch migration in two wild-type ganglionic branches (GBs) at embryonic stage 14/15. Time-lapse images of a wt embryo expressing btl-GAL4UAS-Src-GFP visualized from a ventral view. Images were acquired every min for the time specified (3 h and 20 min) over 50 μm (ZIP 4652 kb)

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© 2023 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

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Araújo, S.J., Llimargas, M. (2023). Time-Lapse Imaging and Morphometric Analysis of Tracheal Development in Drosophila. In: Margadant, C. (eds) Cell Migration in Three Dimensions. Methods in Molecular Biology, vol 2608. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2887-4_11

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  • DOI: https://doi.org/10.1007/978-1-0716-2887-4_11

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-2886-7

  • Online ISBN: 978-1-0716-2887-4

  • eBook Packages: Springer Protocols

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