2D and 3D Quantitative Analysis of Cell Motility and Cytoskeletal Dynamics

2D- and 3D-Dynamic Image Analysis Systems (2D- and 3D-DIAS) for quantitative analysis of cell motility and chemotaxis are described. Particular attention is given to protocols that have proven useful in the quantitation of cell shape changes and pseudopod dynamics during basic cell motility (i.e. crawling in the absence of a chemotactic or other type of extracellular signal) and directed motion. In addition, methods provided, highlight the applicability of this approach to the accurate phenotypic characterizations of cytoskeletal mutations in Dictyostelium discoideum , cytoskeletal alterations in metastatic cells, and cytoskeletal defects in chemotactically defective polymorphonuclear neutrophils.