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Analysis of Focal Adhesion Spatiotemporal Dynamics

Niko Kladt's picture
Workflow Author: 

Mathew E. Berginski, Eric A. Vitriol, Klaus M. Hahn, Shawn M. Gomez

Dimension: 
2D time series
Comment / Instructions: 

The publication describes a quantitative analysis of focal adhesion dynamics that have been imaged using TIRF. All image processing steps are well explained or referenced.

To better understand the dynamic regulation of focal adhesions, we have developed an analysis system for the automated detection, tracking, and data extraction of these structures in living cells. This analysis system was used to quantify the dynamics of fluorescently tagged Paxillin and FAK in NIH 3T3 fibroblasts followed via Total Internal Reflection Fluorescence Microscopy (TIRF). High content time series included the size, shape, intensity, and position of every adhesion present in a living cell. These properties were followed over time, revealing adhesion lifetime and turnover rates, and segregation of properties into distinct zones.

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