Quantifying topography-guided actin dynamics across scales using optical flow
Author
Lee, Rachel MCampanello, Leonard
Hourwitz, Matt J
Alvarez, Phillip
Omidvar, Ava
Fourkas, John T
Losert, Wolfgang
Date
2020-02-05Journal
Molecular Biology of the CellPublisher
American Society for Cell BiologyType
Article
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The dynamic rearrangement of the actin cytoskeleton is an essential component of many mechanotransduction and cellular force generation pathways. Here we use periodic surface topographies with feature sizes comparable to those of in vivo collagen fibers to measure and compare actin dynamics for two representative cell types that have markedly different migratory modes and physiological purposes: slowly migrating epithelial MCF10A cells and polarizing, fast-migrating, neutrophil-like HL60 cells. Both cell types exhibit reproducible guidance of actin waves (esotaxis) on these topographies, enabling quantitative comparisons of actin dynamics. We adapt a computer-vision algorithm, optical flow, to measure the directions of actin waves at the submicron scale. Clustering the optical flow into regions that move in similar directions enables micron-scale measurements of actin-wave speed and direction. Although the speed and morphology of actin waves differ between MCF10A and HL60 cells, the underlying actin guidance by nanotopography is similar in both cell types at the micron and submicron scales.Identifier to cite or link to this item
http://hdl.handle.net/10713/13462ae974a485f413a2113503eed53cd6c53
10.1091/mbc.E19-11-0614
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