High-Resolution Quantification of Focal Adhesion Spatiotemporal Dynamics in Living Cells
(A) One frame from a 200 minute movie of NIH 3T3 cells expressing GFP-Paxillin (the scale bar represents 10 µm). (B) The same cell as in (A), with each adhesion outlined in a different color. (C) The entire set of adhesions in an experiment can be visualized by overlaying the adhesions from each microscopy image using a different color for the set of adhesions at each time point. This example includes the adhesions from 198 images. (D) A range of properties can be extracted from the segmented FA. Five samples are provided. The area histogram was filtered to only include adhesions with areas less than 5 µm2. The axial ratio histogram was filtered to only include adhesions with an axial ratio of 8 or less. The longevity histogram includes all adhesions, while the inset only includes adhesions with longevity greater than 20. The histograms include data from 21 cells.