Journal
NATURE PROTOCOLS
Volume 4, Issue 10, Pages 1397-1412Publisher
NATURE PUBLISHING GROUP
DOI: 10.1038/nprot.2009.130
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Funding
- National Institutes of Health [R01 GM078542]
- Searle Scholars Program
- March of Dimes (Basil O'Conner Starter Scholar Award) [5-FY06-145]
- Caltech Beckman Institute and National Institutes of Health [P50HG004071]
- Caltech Biology Division
- NATIONAL HUMAN GENOME RESEARCH INSTITUTE [P50HG004071] Funding Source: NIH RePORTER
- NATIONAL INSTITUTE OF GENERAL MEDICAL SCIENCES [R01GM078542] Funding Source: NIH RePORTER
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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.
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