title: Multidimensional traction force microscopy reveals out-of-plane rotational moments about focal adhesions.
abstract: Recent methods have revealed that cells on planar substrates exert both shear (in-plane) and normal (out-of-plane) tractions against the extracellular matrix (ECM). However, the location and origin of the normal tractions with respect to the adhesive and cytoskeletal elements of cells have not been elucidated. We developed a high-spatiotemporal-resolution, multidimensional (2.5D) traction force microscopy to measure and model the full 3D nature of cellular forces on planar 2D surfaces. We show that shear tractions are centered under elongated focal adhesions whereas upward and downward normal tractions are detected on distal (toward the cell edge) and proximal (toward the cell body) ends of adhesions, respectively. Together, these forces produce significant rotational moments about focal adhesions in both protruding and retracting peripheral regions. Temporal 2.5D traction force microscopy analysis of migrating and spreading cells shows that these rotational moments are highly dynamic, propagating outward with the leading edge of the cell. Finally, we developed a finite element model to examine how rotational moments could be generated about focal adhesions in a thin lamella. Our model suggests that rotational moments can be generated largely via shear lag transfer to the underlying ECM from actomyosin contractility applied at the intracellular surface of a rigid adhesion of finite thickness. Together, these data demonstrate and probe the origin of a previously unappreciated multidimensional stress profile associated with adhesions and highlight the importance of new approaches to characterize cellular forces.
authors.affiliation: Department of Bioengineering, University of Pennsylvania, Philadelphia, PA 19104, USA.
authors.forename: Wesley R
authors.initials: WR
authors.lastname: Legant
authors.forename: Colin K
authors.initials: CK
authors.lastname: Choi
authors.forename: Jordan S
authors.initials: JS
authors.lastname: Miller
authors.forename: Lin
authors.initials: L
authors.lastname: Shao
authors.forename: Liang
authors.initials: L
authors.lastname: Gao
authors.forename: Eric
authors.initials: E
authors.lastname: Betzig
authors.forename: Christopher S
authors.initials: CS
authors.lastname: Chen
mesh_terms: D000818:Animals; D055592:Biophysical Phenomena; D005347:Fibroblasts / Q000166:cytology / Q000502:physiology; D020342:Finite Element Analysis; D022001:Focal Adhesions / Q000502:physiology*; D049452:Green Fluorescent Proteins / Q000378:metabolism; D040542:Mechanotransduction, Cellular / Q000502:physiology; D051379:Mice; D008856:Microscopy, Fluorescence; D008954:Models, Biological; D011994:Recombinant Proteins / Q000378:metabolism; D012399:Rotation
chemical_list: D011994:Recombinant Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins