Extracting subcellular structural dynamics from multi-wavelength 4-D fluorescence images

Endothelial cells lining the arteries remodel the extracellular matrix (ECM) in regions of disturbed blood flow during development of atherosclerosis, but mechanisms remain unclear. Multi-wavelength 4-D fluorescence images of adhesion and ECM proteins in endothelial cells were acquired during changes in fluid shear stress. Using centroid coordinates of focal adhesion sites, Voronoi polygons were generated such that interior points were associated with the closest adhesion site. Image segmentation, block matching, and tracking algorithms enabled displacement field estimation for both focal adhesions and associated ECM structures. This quantitative image analysis approach reveals structural dynamics during flow-mediated ECM and adhesion site remodeling.