Supatto, Willy and McMahon, Amy and Fraser, Scott E. and Stathopoulos, Angelike (2009) Quantitative imaging of collective cell migration during Drosophila gastrulation: multiphoton microscopy and computational analysis. Nature Protocols, 4 (10). pp. 1397-1412. ISSN 1754-2189 http://resolver.caltech.edu/CaltechAUTHORS:20100414-093601075
![[img]](http://authors.library.caltech.edu/style/images/fileicons/application_pdf.png) |
PDF
- Published Version
Restricted to Repository administrators only See Usage Policy. 812Kb |
![[img]](http://authors.library.caltech.edu/style/images/fileicons/application_zip.png) |
Archive (ZIP) (Supplementary Data 1. Matlab scripts.)
- Supplemental Material
Restricted to Repository administrators only See Usage Policy. 15Kb |
|
Archive (ZIP) (Supplementary Data 2. Sample datasets.)
- Supplemental Material
Restricted to Repository administrators only See Usage Policy. 801Kb |
![[img]](http://authors.library.caltech.edu/style/images/fileicons/video.png) |
Video (QuickTime) (Supplementary Movie 1: 3D image of an entire klarsicht;nls-GFP embryo at the onset of mesoderm cell spreading)
- Supplemental Material
Restricted to Repository administrators only See Usage Policy. 2754Kb |
|
Video (QuickTime) (Supplementary Movie 2: 3D schematic animation of quantitative imaging of Drosophila embryos)
- Supplemental Material
Restricted to Repository administrators only See Usage Policy. 6Mb |
|
Video (QuickTime) (Supplementary Movie 3: Example of angular drift during image acquisition)
- Supplemental Material
Restricted to Repository administrators only See Usage Policy. 3432Kb |
Use this Persistent URL to link to this item: http://resolver.caltech.edu/CaltechAUTHORS:20100414-093601075
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.
| Item Type: | Article | ||||||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| 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. | ||||||||||||||
| Funders: |
| ||||||||||||||
| Subject Keywords: | Cell and developmental biology; Computational and theoretical biology; Imaging; Model organisms | ||||||||||||||
| Record Number: | CaltechAUTHORS:20100414-093601075 | ||||||||||||||
| Persistent URL: | http://resolver.caltech.edu/CaltechAUTHORS:20100414-093601075 | ||||||||||||||
| Related URLs: | |||||||||||||||
| Official Citation: | Supatto, W., A. McMahon, et al. (2009). "Quantitative imaging of collective cell migration during Drosophila gastrulation: multiphoton microscopy and computational analysis." Nat. Protocols 4(10): 1397-1412. | ||||||||||||||
| Usage Policy: | No commercial reproduction, distribution, display or performance rights in this work are provided. | ||||||||||||||
| ID Code: | 17973 | ||||||||||||||
| Collection: | CaltechAUTHORS | ||||||||||||||
| Deposited By: | Jason Perez | ||||||||||||||
| Deposited On: | 14 May 2010 18:09 | ||||||||||||||
| Last Modified: | 26 Dec 2012 11:58 |
Repository Staff Only: item control page
