title: Three-dimensional quantification of cellular traction forces and mechanosensing of thin substrata by fourier traction force microscopy.
abstract: We introduce a novel three-dimensional (3D) traction force microscopy (TFM) method motivated by the recent discovery that cells adhering on plane surfaces exert both in-plane and out-of-plane traction stresses. We measure the 3D deformation of the substratum on a thin layer near its surface, and input this information into an exact analytical solution of the elastic equilibrium equation. These operations are performed in the Fourier domain with high computational efficiency, allowing to obtain the 3D traction stresses from raw microscopy images virtually in real time. We also characterize the error of previous two-dimensional (2D) TFM methods that neglect the out-of-plane component of the traction stresses. This analysis reveals that, under certain combinations of experimental parameters (cell size, substratums' thickness and Poisson's ratio), the accuracy of 2D TFM methods is minimally affected by neglecting the out-of-plane component of the traction stresses. Finally, we consider the cell's mechanosensing of substratum thickness by 3D traction stresses, finding that, when cells adhere on thin substrata, their out-of-plane traction stresses can reach four times deeper into the substratum than their in-plane traction stresses. It is also found that the substratum stiffness sensed by applying out-of-plane traction stresses may be up to 10 times larger than the stiffness sensed by applying in-plane traction stresses.
authors.affiliation: Mechanical and Aerospace Engineering Department, University of California San Diego, La Jolla, California, United States of America ; Institute for Engineering in Medicine, University of California San Diego, La Jolla, California, United States of America.
authors.forename: Juan C
authors.initials: JC
authors.lastname: del Álamo
authors.forename: Ruedi
authors.initials: R
authors.lastname: Meili
authors.forename: Begoña
authors.initials: B
authors.lastname: Álvarez-González
authors.forename: Baldomero
authors.initials: B
authors.lastname: Alonso-Latorre
authors.forename: Effie
authors.initials: E
authors.lastname: Bastounis
authors.forename: Richard
authors.initials: R
authors.lastname: Firtel
authors.forename: Juan C
authors.initials: JC
authors.lastname: Lasheras
mesh_terms: D004023:Dictyostelium; D008853:Microscopy / Q000379:methods*
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