Abstract
Laser ablation is nowadays a widespread technique to probe tissue mechanics during development. Here we describe the setup of one such ablation system and ablation experiments performed on the embryo and pupa of Drosophila. We describe in detail the process of sample preparation, how to disrupt single-cell junctions and perform linear or circular cuts at the tissue scale, and how to analyze the data to determine relevant mechanical parameters.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Heisenberg C-P, Bellaïche Y (2013) Forces in tissue morphogenesis and patterning. Cell 153:948–962
Hutson MS, Tokutake Y et al (2003) Forces for morphogenesis investigated with laser microsurgery and quantitative modeling. Science 300:145–149
Farhadifar F, Roper J-C, Aigouy B et al (2007) The influence of cell mechanics, cell-cell interactions, and proliferation on epithelial packing. Curr Biol 17:2095–2104
Rauzi M, Verant P, Lecuit T et al (2008) Nature and anisotropy of cortical forces orienting Drosophila tissue morphogenesis. Nat Cell Biol 10:1401–1410
Rauzi M, Lenne P-F (2011) Cortical forces in cell shape changes and tissue morphogenesis. Curr Top Dev Biol. doi:10.1016/B978-0-12-385065-2.00004-9
Bosveld F, Bonnet I, Guirao B et al (2012) Mechanical control of morphogenesis by Fat/Dachsous/Four-Jointed Planar Cell Polarity Pathway. Science. doi:10.1126/science.1221071
Bonnet I, Marcq P (2012) Mechanical state, material properties and continuous description of an epithelial tissue. J R Soc Interface. doi:10.1098/rsif.2012.0263
Cavey M, Lecuit T (2008) Imaging cellular and molecular dynamics in live embryos using fluorescent proteins. Methods Mol Biol. doi:10.1007/978-1-59745-542-8_8
Kiehart DP, Galbraith CG, Edwards KA et al (2000) Multiple forces contribute to cell sheet morphogenesis for dorsal closure in Drosophila. J Cell Biol 149:471–490
Acknowledgments
We thank all members of the Lenne and Lecuit groups for discussions and comments on the manuscript, in particular Claire Chardès and Crystal Love. This work has been supported by the Fondation de la Recherche Médicale (DEQ20130326509) and the Labex INFORM (ANR-11-LABX-0054) and of the A*MIDEX project (ANR-11-IDEX-0001-02), funded by the « Investissements d’Avenir » French Government program. PS is supported by a PhD fellowship from the Labex INFORM.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2016 Springer Science+Business Media New York
About this protocol
Cite this protocol
Shivakumar, P.C., Lenne, PF. (2016). Laser Ablation to Probe the Epithelial Mechanics in Drosophila . In: Dahmann, C. (eds) Drosophila. Methods in Molecular Biology, vol 1478. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6371-3_14
Download citation
DOI: https://doi.org/10.1007/978-1-4939-6371-3_14
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-6369-0
Online ISBN: 978-1-4939-6371-3
eBook Packages: Springer Protocols