5. Set the time interval and total duration of the movie (see
Note 11).
6. Start the acquisition.
7. At the end of the movie, cells can be fixed in formaldehyde 4%
for further analysis and staining (see Note 12). Typical dispersion of control neural crest explants is shown in Movie 4 and
Fig. 4.
Fig. 4 Analyzing the dynamics of cell dispersion. (a) Workflow from cell culture to substacks for dispersion
analyses. (b, c) Total explored area is retrieved by linking all external cells per explant, per time point. (d–f)
Triangulation analysis. (d) Thresholded image of DAPI staining. (e) Triangles obtained after performing
particles detection and Delaunay/Voronoi triangulation. Each triangle is color-coded according to the following
ratio (area of the triangle/total area). (f) Example of artifacts in areas of high cell density due to poor particle
detection. Steps on panels (d) and (e) are repeated for each time point of interest
Using Xenopus Neural Crest to Study EMT
265
Note 11).
6. Start the acquisition.
7. At the end of the movie, cells can be fixed in formaldehyde 4%
for further analysis and staining (see Note 12). Typical dispersion of control neural crest explants is shown in Movie 4 and
Fig. 4.
Fig. 4 Analyzing the dynamics of cell dispersion. (a) Workflow from cell culture to substacks for dispersion
analyses. (b, c) Total explored area is retrieved by linking all external cells per explant, per time point. (d–f)
Triangulation analysis. (d) Thresholded image of DAPI staining. (e) Triangles obtained after performing
particles detection and Delaunay/Voronoi triangulation. Each triangle is color-coded according to the following
ratio (area of the triangle/total area). (f) Example of artifacts in areas of high cell density due to poor particle
detection. Steps on panels (d) and (e) are repeated for each time point of interest
Using Xenopus Neural Crest to Study EMT
265