Protocol abstract
Nature Protocols 4, - 1397 - 1412 (2009)
Published online: 10 September 2009 | doi:10.1038/nprot.2009.130
Subject Categories: Cell and developmental biology | Computational and theoretical biology | Imaging | Model organisms
Quantitative imaging of collective cell migration during Drosophila gastrulation: multiphoton microscopy and computational analysis
Willy Supatto1,2, Amy McMahon1, Scott E Fraser1,2 & Angelike Stathopoulos1
Abstract
This protocol describes imaging and computational tools to collect and analyze live imaging data of embryonic cell migration. Our five-step protocol requires a few weeks to move through embryo preparation and four-dimensional (4D) live imaging using multi-photon microscopy, to 3D cell tracking using image processing, registration of tracking data and their quantitative analysis using computational tools. It uses commercially available equipment and requires expertise in microscopy and programming that is appropriate for a biology laboratory. Custom-made scripts are provided, as well as sample datasets to permit readers without experimental data to carry out the analysis. The protocol has offered new insights into the genetic control of cell migration during Drosophila gastrulation. With simple modifications, this systematic analysis could be applied to any developing system to define cell positions in accordance with the body plan, to decompose complex 3D movements and to quantify the collective nature of cell migration.
- Division of Biology and Beckman Institute, California Institute of Technology, Pasadena, California, USA.
- Present address: Institut Jacques Monod, CNRS UMR 7592, Université Paris 7, Paris, France.
Correspondence to: Willy Supatto1,2 e-mail: supatto@caltech.edu
Correspondence to: Angelike Stathopoulos1 e-mail: angelike@caltech.edu
nature-products
A-Z product listing
- 2PEF Microscope(Coherent)
- Bleach (Clorox) or sodium hypochlorite(Sigma)
- C-Aprochromat 40
/1.1 NA W Corr UV-VIS-IR(Carl Zeiss) - cell strainers(BD Falcon)
- Coverslips(VWR)
- Double-sided sticky tape(TESA)
