Quantitative imaging of collective cell migration during Drosophila gastrulation: multiphoton microscopy and computational analysis
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1.
California Institute of Technology
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 multiphoton 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.
Additional Information
© 2010 Nature Publishing Group. Published online: 10 September 2009. We thank M. Liebling for advice on Imaris and Matlab, and the Caltech Biological Imaging Center for sharing equipment. This work was supported by grants to AS from National Institutes of Health (R01 GM078542), the Searle Scholars Program and the March of Dimes (Basil O'Conner Starter Scholar Award, 5-FY06-145), grants to SF from the Caltech Beckman Institute and National Institutes of Health (Center for Excellence in Genomic Science grant P50HG004071), and fellowship to W.S. from the Caltech Biology Division. W.S. designed the experimental workflow and the computational analysis. W.S. and A.M. acquired and processed the data. W.S., A.M., S.E.F. and A.S. wrote the paper.Attached Files
Accepted Version - nihms-163224.pdf
Supplemental Material - Supplementary_Movie_1.mov
Supplemental Material - Supplementary_Movie_2.mov
Supplemental Material - Supplementary_Movie_3.mov
Supplemental Material - nprot.2009.130-SI1.zip
Supplemental Material - nprot.2009.130-SI2.zip
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Additional details
- PMCID
- PMC2854020
- Eprint ID
- 17973
- DOI
- 10.1038/nprot.2009.130
- Resolver ID
- CaltechAUTHORS:20100414-093601075
- NIH
- R01 GM078542
- Searle Scholars Program
- March of Dimes
- 5-FY06-145
- Caltech Beckman Institute
- NIH
- P50HG004071
- Caltech Biology Division
- Created
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2010-05-14Created from EPrint's datestamp field
- Updated
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2021-11-08Created from EPrint's last_modified field