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

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.

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

2020 ◽  
Vol 4 (3) ◽  
pp. 036104 ◽  
Author(s):  
Elisabeth E. Charrier ◽  
Katarzyna Pogoda ◽  
Robin Li ◽  
Chan Young Park ◽  
Jeffrey J. Fredberg ◽  
...  

Author(s):  
Steven Huth ◽  
Johannes W. Blumberg ◽  
Dimitri Probst ◽  
Jan Lammerding ◽  
Ulrich S. Schwarz ◽  
...  

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.


2021 ◽  
Vol 32 (18) ◽  
pp. 1737-1748
Author(s):  
Somanna Kollimada ◽  
Fabrice Senger ◽  
Timothée Vignaud ◽  
Manuel Théry ◽  
Laurent Blanchoin ◽  
...  

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.


2020 ◽  
Vol 30 (1) ◽  
pp. 74-83 ◽  
Author(s):  
Abigail De La Pena ◽  
Marah Mukhtar ◽  
Ryosuke Yokosawa ◽  
Santiago Carrasquilla ◽  
Chelsey S. Simmons

2020 ◽  
Author(s):  
Jorge Barrasa Fano ◽  
Apeksha Shapeti ◽  
Alvaro Jorge-Penas ◽  
Mojtaba Barzegari ◽  
Jose A Sanz Herrera ◽  
...  

We present TFMLAB, a MATLAB software package for 4D (x;y;z;t) Traction Force Microscopy (TFM). While various TFM computational workflows are available in the literature, open-source programs that are easy to use by researchers with limited technical experience and that can analyze 4D in vitro systems do not exist. TFMLAB integrates all the computational steps to compute active cellular forces from confocal microscopy images, including image processing, cell segmentation, image alignment, matrix displacement measurement and force recovery. Moreover, TFMLAB eases usability by means of interactive graphical user interfaces. This work describes the package's functionalities and analyses its performance on a real TFM case.


2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Liliana Barbieri ◽  
Huw Colin-York ◽  
Kseniya Korobchevskaya ◽  
Di Li ◽  
Deanna L. Wolfson ◽  
...  

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.


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