Open Access Methodology article

Automated characterization of cell shape changes during amoeboid motility by skeletonization

Yuan Xiong1, Cathryn Kabacoff2, Jonathan Franca-Koh2, Peter N Devreotes2, Douglas N Robinson2 and Pablo A Iglesias1*

  • * Corresponding author: Pablo A Iglesias

Author Affiliations

1 Department of Electrical and Computer Engineering, Johns Hopkins University, Baltimore, MD 21218 USA

2 Department of Cell Biology, Johns Hopkins University School of Medicine, Baltimore, MD 21205 USA

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BMC Systems Biology 2010, 4:33  doi:10.1186/1752-0509-4-33

Published: 24 March 2010



The ability of a cell to change shape is crucial for the proper function of many cellular processes, including cell migration. One type of cell migration, referred to as amoeboid motility, involves alternating cycles of morphological expansion and retraction. Traditionally, this process has been characterized by a number of parameters providing global information about shape changes, which are insufficient to distinguish phenotypes based on local pseudopodial activities that typify amoeboid motility.


We developed a method that automatically detects and characterizes pseudopodial behavior of cells. The method uses skeletonization, a technique from morphological image processing to reduce a shape into a series of connected lines. It involves a series of automatic algorithms including image segmentation, boundary smoothing, skeletonization and branch pruning, and takes into account the cell shape changes between successive frames to detect protrusion and retraction activities. In addition, the activities are clustered into different groups, each representing the protruding and retracting history of an individual pseudopod.


We illustrate the algorithms on movies of chemotaxing Dictyostelium cells and show that our method makes it possible to capture the spatial and temporal dynamics as well as the stochastic features of the pseudopodial behavior. Thus, the method provides a powerful tool for investigating amoeboid motility.