Chapter 17
Multiscale In Vivo Imaging of Collective Cell Migration
in Drosophila Embryos
Gordana Scepanovic, Alexandru Florea, and Rodrigo Fernandez-Gonzalez
Abstract
Coordinated cell movements drive embryonic development and tissue repair, and can also spread disease.
Time-lapse microscopy is an integral part in the study of the cell biology of collective cell movements.
Advances in imaging techniques enable monitoring dynamic cellular and molecular events in real time
within living animals. Here, we demonstrate the use of spinning disk confocal microscopy to investigate
coordinated cell movements and epithelial-to-mesenchymal-like transitions during embryonic wound
closure in Drosophila. We describe image-based metrics to quantify the efficiency of collective cell migration.
Finally, we show the application of super-resolution radial fluctuation microscopy to obtain multidimensional, super-resolution images of protrusive activity in collectively moving cells in vivo. Together, the
methods presented here constitute a toolkit for the modern analysis of collective cell migration in living
animals.
Key words Drosophila melanogaster, Live imaging, Mounting methods, Quantitative microscopy,
Spinning disk confocal microscopy, Super-resolution radial fluctuation microscopy, Wound healing
1 Introduction
Collective cell movements are essential for embryonic development. Morphogenetic processes such as gastrulation [1] and limb
elongation [2] are mediated by coordinated cell movements and
failure of cells to organize cell migration can lead to congenital
birth defects. Coordinated cell movements also contribute to the
spread of disease during cancer invasion [3]. Therefore, understanding the mechanisms by which cells coordinate their movements may facilitate the development of therapeutic interventions
that promote or prevent collective cell migration.
Kyra Campbell and Eric Theveneau (eds.), The Epithelial-to Mesenchymal Transition: Methods and Protocols,
Methods in Molecular Biology, vol. 2179, https://doi.org/10.1007/978-1-0716-0779-4_17,
© Springer Science+Business Media, LLC, part of Springer Nature 2021
Electronic supplementary material: The online version of this chapter (https://doi.org/10.1007/978-1-07160779-4_17) contains supplementary material, which is available to authorized users.
199
Multiscale In Vivo Imaging of Collective Cell Migration
in Drosophila Embryos
Gordana Scepanovic, Alexandru Florea, and Rodrigo Fernandez-Gonzalez
Abstract
Coordinated cell movements drive embryonic development and tissue repair, and can also spread disease.
Time-lapse microscopy is an integral part in the study of the cell biology of collective cell movements.
Advances in imaging techniques enable monitoring dynamic cellular and molecular events in real time
within living animals. Here, we demonstrate the use of spinning disk confocal microscopy to investigate
coordinated cell movements and epithelial-to-mesenchymal-like transitions during embryonic wound
closure in Drosophila. We describe image-based metrics to quantify the efficiency of collective cell migration.
Finally, we show the application of super-resolution radial fluctuation microscopy to obtain multidimensional, super-resolution images of protrusive activity in collectively moving cells in vivo. Together, the
methods presented here constitute a toolkit for the modern analysis of collective cell migration in living
animals.
Key words Drosophila melanogaster, Live imaging, Mounting methods, Quantitative microscopy,
Spinning disk confocal microscopy, Super-resolution radial fluctuation microscopy, Wound healing
1 Introduction
Collective cell movements are essential for embryonic development. Morphogenetic processes such as gastrulation [1] and limb
elongation [2] are mediated by coordinated cell movements and
failure of cells to organize cell migration can lead to congenital
birth defects. Coordinated cell movements also contribute to the
spread of disease during cancer invasion [3]. Therefore, understanding the mechanisms by which cells coordinate their movements may facilitate the development of therapeutic interventions
that promote or prevent collective cell migration.
Kyra Campbell and Eric Theveneau (eds.), The Epithelial-to Mesenchymal Transition: Methods and Protocols,
Methods in Molecular Biology, vol. 2179, https://doi.org/10.1007/978-1-0716-0779-4_17,
© Springer Science+Business Media, LLC, part of Springer Nature 2021
Electronic supplementary material: The online version of this chapter (https://doi.org/10.1007/978-1-07160779-4_17) contains supplementary material, which is available to authorized users.
199