scholarly journals Quantifying force transmission through fibroblasts: changes of traction forces under external shearing

2021 ◽  
Author(s):  
Steven Huth ◽  
Johannes W. Blumberg ◽  
Dimitri Probst ◽  
Jan Lammerding ◽  
Ulrich S. Schwarz ◽  
...  
Keyword(s):  
Cell Surface ◽  
Mammalian Cells ◽  
Shear Force ◽  
Apical Cell ◽  
Traction Force ◽  
Traction Force Microscopy ◽  
Force Transmission ◽  
Traction Forces ◽  
Force Microscopy ◽  
Adhesion Sites

AbstractMammalian cells have evolved complex mechanical connections to their microenvironment, including focal adhesion clusters that physically connect the cytoskeleton and the extracellular matrix. This mechanical link is also part of the cellular machinery to transduce, sense and respond to external forces. Although methods to measure cell attachment and cellular traction forces are well established, these are not capable of quantifying force transmission through the cell body to adhesion sites. We here present a novel approach to quantify intracellular force transmission by combining microneedle shearing at the apical cell surface with traction force microscopy at the basal cell surface. The change of traction forces exerted by fibroblasts to underlying polyacrylamide substrates as a response to a known shear force exerted with a calibrated microneedle reveals that cells redistribute forces dynamically under external shearing and during sequential rupture of their adhesion sites. Our quantitative results demonstrate a transition from dipolar to monopolar traction patterns, an inhomogeneous distribution of the external shear force to the adhesion sites as well as dynamical changes in force loading prior to and after the rupture of single adhesion sites. Our strategy of combining traction force microscopy with external force application opens new perspectives for future studies of force transmission and mechanotransduction in cells.

scholarly journals pyTFM: A tool for traction force and monolayer stress microscopy

2021 ◽  
Vol 17 (6) ◽  
pp. e1008364
Author(s):  
Andreas Bauer ◽  
Magdalena Prechová ◽  
Lena Fischer ◽  
Ingo Thievessen ◽  
Martin Gregor ◽  
...  
Keyword(s):  
Image Annotation ◽  
Traction Force ◽  
Theoretical Background ◽  
Ease Of Use ◽  
Traction Force Microscopy ◽  
Force Transmission ◽  
Force Microscopy ◽  
Wide Range ◽  
Cell Layers ◽  
Cellular Stresses

Cellular force generation and force transmission are of fundamental importance for numerous biological processes and can be studied with the methods of Traction Force Microscopy (TFM) and Monolayer Stress Microscopy. Traction Force Microscopy and Monolayer Stress Microscopy solve the inverse problem of reconstructing cell-matrix tractions and inter- and intra-cellular stresses from the measured cell force-induced deformations of an adhesive substrate with known elasticity. Although several laboratories have developed software for Traction Force Microscopy and Monolayer Stress Microscopy computations, there is currently no software package available that allows non-expert users to perform a full evaluation of such experiments. Here we present pyTFM, a tool to perform Traction Force Microscopy and Monolayer Stress Microscopy on cell patches and cell layers grown in a 2-dimensional environment. pyTFM was optimized for ease-of-use; it is open-source and well documented (hosted at https://pytfm.readthedocs.io/) including usage examples and explanations of the theoretical background. pyTFM can be used as a standalone Python package or as an add-on to the image annotation tool ClickPoints. In combination with the ClickPoints environment, pyTFM allows the user to set all necessary analysis parameters, select regions of interest, examine the input data and intermediary results, and calculate a wide range of parameters describing forces, stresses, and their distribution. In this work, we also thoroughly analyze the accuracy and performance of the Traction Force Microscopy and Monolayer Stress Microscopy algorithms of pyTFM using synthetic and experimental data from epithelial cell patches.

scholarly journals A Novel Method for Quantifying Traction Forces from Hexagonal Micropatterned Features on Deformable Poly-Dimethyl Siloxane Sheets

2018 ◽  
Author(s):  
Brian P. Griffin ◽  
Christopher J. Largaespada ◽  
Nicole A. Rinaldi ◽  
Christopher A. Lemmon
Keyword(s):  
Computational Algorithm ◽  
Traction Force ◽  
Traction Force Microscopy ◽  
Polydimethyl Siloxane ◽  
Traction Forces ◽  
Homogeneous Surface ◽  
Force Microscopy ◽  
Dimethyl Siloxane ◽  
Novel Method ◽  
Cellular Forces

AbstractMany methods exist for quantifying cellular traction forces, including traction force microscopy and microfabricated post arrays. However, these methodologies have limitations, including a requirement to remove cells to determine undeflected particle locations and the inability to quantify forces of cells with low cytoskeletal stiffness, respectively. Here we present a novel method of traction force quantification that eliminates both of these limitations. Through the use of a hexagonal pattern of microcontact-printed protein spots, a novel computational algorithm, and thin surfaces of polydimethyl siloxane (PDMS) blends, we demonstrate a system that quantifies cellular forces on a homogeneous surface that is stable, easily manufactured, and can quantify forces without need for cellular removal.

scholarly journals Three-Dimensional Quantification of Cellular Traction Forces and Mechanosensing of Thin Substrata by Fourier Traction Force Microscopy

PLoS ONE ◽  
2013 ◽  
Vol 8 (9) ◽  
pp. e69850 ◽  
Author(s):  
Juan C. del Álamo ◽  
Ruedi Meili ◽  
Begoña Álvarez-González ◽  
Baldomero Alonso-Latorre ◽  
Effie Bastounis ◽  
...  
Keyword(s):  
Three Dimensional ◽  
Traction Force ◽  
Traction Force Microscopy ◽  
Traction Forces ◽  
Force Microscopy

scholarly journals The biochemical composition of the actomyosin network sets the magnitude of cellular traction forces

2021 ◽  
Vol 32 (18) ◽  
pp. 1737-1748
Author(s):  
Somanna Kollimada ◽  
Fabrice Senger ◽  
Timothée Vignaud ◽  
Manuel Théry ◽  
Laurent Blanchoin ◽  
...  
Keyword(s):  
Focal Adhesion ◽  
Biochemical Composition ◽  
Force Production ◽  
Traction Force ◽  
Traction Force Microscopy ◽  
Traction Forces ◽  
Force Microscopy ◽  
Cell Force

The endogenous content of proteins associated with force production and the resultant traction forces were quantified in the same cells using a new traction force-microscopy assay. Focal adhesion size correlated with force in stationary cells. Relative numbers of motors and cross-linkers per actin required an optimum to maximize cell force production.

scholarly journals Two-dimensional TIRF-SIM–traction force microscopy (2D TIRF-SIM-TFM)

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Liliana Barbieri ◽  
Huw Colin-York ◽  
Kseniya Korobchevskaya ◽  
Di Li ◽  
Deanna L. Wolfson ◽  
...  
Keyword(s):  
Resolution Enhancement ◽  
Traction Force ◽  
Super Resolution ◽  
Traction Force Microscopy ◽  
Two Dimensional ◽  
Structured Illumination ◽  
Structured Illumination Microscopy ◽  
Force Microscopy ◽  
Health And Disease ◽  
Spatio Temporal

AbstractQuantifying small, rapidly evolving forces generated by cells is a major challenge for the understanding of biomechanics and mechanobiology in health and disease. Traction force microscopy remains one of the most broadly applied force probing technologies but typically restricts itself to slow events over seconds and micron-scale displacements. Here, we improve >2-fold spatially and >10-fold temporally the resolution of planar cellular force probing compared to its related conventional modalities by combining fast two-dimensional total internal reflection fluorescence super-resolution structured illumination microscopy and traction force microscopy. This live-cell 2D TIRF-SIM-TFM methodology offers a combination of spatio-temporal resolution enhancement relevant to forces on the nano- and sub-second scales, opening up new aspects of mechanobiology to analysis.

scholarly journals Different Cell Migration Modes: Insights from Traction Force Microscopy and Modeling

2021 ◽  
Vol 120 (3) ◽  
pp. 113a
Author(s):  
Wouter-Jan Rappel ◽  
Elisabeth Ghabache ◽  
Yuansheng Cao ◽  
Yuchuan Miao ◽  
Alexander Groisman ◽  
...  
Keyword(s):  
Cell Migration ◽  
Traction Force ◽  
Traction Force Microscopy ◽  
Force Microscopy

Cell surface interactions in conjugation: Tetrahymena ciliary membrane vesicles

1984 ◽  
Vol 4 (4) ◽  
pp. 681-687
Author(s):  
B Love ◽  
M B Rotheim
Keyword(s):  
Cell Surface ◽  
Mating Type ◽  
Mammalian Cells ◽  
Membrane Vesicles ◽  
Surface Interactions ◽  
Dependent Manner ◽  
Concentration Dependent Manner ◽  
Ciliary Membrane ◽  
Adhesion Sites ◽  
Cell Surface Interactions

Tetrahymena ciliary membrane vesicles are shown to interact with preconjugant cells in a mating type-specific way. When cells are treated with vesicles of a different mating type before mixing for conjugation, cell pairing is enhanced, and the normal prepairing period is partially eliminated. This enhancement is mating type specific since it is not observed after pretreatment of cells with vesicles of their own mating type. In contrast, when vesicles are added at the time of mixing of two starved cultures, cell pairing is delayed in a concentration-dependent manner. By varying the conditions, we demonstrated enhancement or inhibition, or both. These results are interpreted in terms of two independent interactions of cells with vesicles. We suggest that first, vesicles substitute for another cell in cell-cell prepairing interaction and second, vesicles compete for adhesion sites produced during the prepairing period. Finally, the data presented are summarized within a speculative framework that calls attention to potential analogies with hormone-receptor signaling in mammalian cells.

scholarly journals Epifluorescence-based three-dimensional traction force microscopy

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Lauren Hazlett ◽  
Alexander K. Landauer ◽  
Mohak Patel ◽  
Hadley A. Witt ◽  
Jin Yang ◽  
...  
Keyword(s):  
Spatial Frequency ◽  
Open Source ◽  
Single Particle ◽  
Three Dimensional ◽  
Traction Force ◽  
High Spatial Frequency ◽  
Epifluorescence Microscopy ◽  
Traction Force Microscopy ◽  
Force Microscopy ◽  
Out Of Plane

Abstract We introduce a novel method to compute three-dimensional (3D) displacements and both in-plane and out-of-plane tractions on nominally planar transparent materials using standard epifluorescence microscopy. Despite the importance of out-of-plane components to fully understanding cell behavior, epifluorescence images are generally not used for 3D traction force microscopy (TFM) experiments due to limitations in spatial resolution and measuring out-of-plane motion. To extend an epifluorescence-based technique to 3D, we employ a topology-based single particle tracking algorithm to reconstruct high spatial-frequency 3D motion fields from densely seeded single-particle layer images. Using an open-source finite element (FE) based solver, we then compute the 3D full-field stress and strain and surface traction fields. We demonstrate this technique by measuring tractions generated by both single human neutrophils and multicellular monolayers of Madin–Darby canine kidney cells, highlighting its acuity in reconstructing both individual and collective cellular tractions. In summary, this represents a new, easily accessible method for calculating fully three-dimensional displacement and 3D surface tractions at high spatial frequency from epifluorescence images. We released and support the complete technique as a free and open-source code package.

scholarly journals Traction Force Microscopy Based on an Active Cable Network Model

2014 ◽  
Vol 106 (2) ◽  
pp. 425a
Author(s):  
Jerome Soine ◽  
Christoph Brand ◽  
Jonathan Stricker ◽  
Patrick W. Oakes ◽  
Margaret L. Gardel ◽  
...  
Keyword(s):  
Network Model ◽  
Traction Force ◽  
Traction Force Microscopy ◽  
Force Microscopy ◽  
Cable Network
Sign in / Sign up

Export Citation Format

Share Document