scholarly journals Collective forces of tumor spheroids in three-dimensional biopolymer networks

eLife ◽  
2020 ◽  
Vol 9 ◽  
Author(s):  
Christoph Mark ◽  
Thomas J Grundy ◽  
Pamela L Strissel ◽  
David Böhringer ◽  
Nadine Grummel ◽  
...  
Keyword(s):  
Single Cells ◽  
Three Dimensional ◽  
Traction Force ◽  
Tumor Spheroids ◽  
Scale Invariant ◽  
Tissue Samples ◽  
Force Microscopy ◽  
Surrounding Matrix ◽  
Highly Nonlinear ◽  
Contractile Forces

We describe a method for quantifying the contractile forces that tumor spheroids collectively exert on highly nonlinear three-dimensional collagen networks. While three-dimensional traction force microscopy for single cells in a nonlinear matrix is computationally complex due to the variable cell shape, here we exploit the spherical symmetry of tumor spheroids to derive a scale-invariant relationship between spheroid contractility and the surrounding matrix deformations. This relationship allows us to directly translate the magnitude of matrix deformations to the total contractility of arbitrarily sized spheroids. We show that our method is accurate up to strains of 50% and remains valid even for irregularly shaped tissue samples when considering only the deformations in the far field. Finally, we demonstrate that collective forces of tumor spheroids reflect the contractility of individual cells for up to 1 hr after seeding, while collective forces on longer timescales are guided by mechanical feedback from the extracellular matrix.

scholarly journals Collective forces of tumor spheroids in three-dimensional biopolymer networks

2019 ◽  
Author(s):  
Christoph Mark ◽  
Thomas J. Grundy ◽  
David Böhringer ◽  
Julian Steinwachs ◽  
Geraldine M. O’Neill ◽  
...  
Keyword(s):  
Single Cells ◽  
Three Dimensional ◽  
Traction Force ◽  
Tumor Spheroids ◽  
Scale Invariant ◽  
Force Microscopy ◽  
Surrounding Matrix ◽  
Highly Nonlinear ◽  
Nonlinear Matrix ◽  
Contractile Forces

ABSTRACTWe describe a method for quantifying the contractile forces that tumor spheroids collectively exert on highly nonlinear three-dimensional collagen networks. While three-dimensional traction force microscopy for single cells in a nonlinear matrix is computationally complex due to the variable cell shape, here we exploit the spherical symmetry of tumor spheroids to derive a scale-invariant relationship between spheroid contractility and the surrounding matrix deformations. This relationship allows us to directly translate the magnitude of matrix deformations to the total contractility of arbitrarily sized spheroids. We show that collective forces of tumor spheroids reflect the contractility of individual cells for up to 1h after seeding, while collective forces on longer time-scales are guided by mechanical feedback from the extracellular matrix.

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 Three-dimensional Traction Force Microscopy for Studying Cellular Interactions with Biomaterials

2012 ◽  
Vol 4 ◽  
pp. 144-150 ◽  
Author(s):  
J. Notbohm ◽  
J.-H. Kim ◽  
C. Franck ◽  
S. Maskarinec ◽  
D. Tirrell ◽  
...  
Keyword(s):  
Three Dimensional ◽  
Traction Force ◽  
Traction Force Microscopy ◽  
Cellular Interactions ◽  
Force Microscopy

Application of 3D Traction Force Microscopy to Mechanotransduction of Cell Clusters

2011 ◽  
Vol 70 ◽  
pp. 21-27 ◽  
Author(s):  
Jacob Notbohm ◽  
Jin Hong Kim ◽  
Anand Asthagiri ◽  
Guruswami Ravichandran
Keyword(s):  
Finite Element ◽  
Three Dimensional ◽  
Traction Force ◽  
Two Dimensions ◽  
Three Dimensions ◽  
Cell Contact ◽  
Traction Force Microscopy ◽  
Inhibition Of Proliferation ◽  
Force Microscopy ◽  
Cell Clusters

With increasing understanding of the important role mechanics plays in cell behavior, the experimental technique of traction force microscopy has grown in popularity over the past decade. While researchers have assumed that cells on a flat substrate apply tractions in only two dimensions, a finite element simulation is discussed here that demonstrates how cells apply tractions in all three dimensions. Three dimensional traction force microscopy is then used to experimentally confirm the finite element results. Finally, the implications that the traction distributions of cell clusters have on the study of inhibition of proliferation due to cell contact and scattering of cells in a cluster are discussed.

scholarly journals Three-Dimensional Traction Force Microscopy: A New Tool for Quantifying Cell-Matrix Interactions

PLoS ONE ◽  
2011 ◽  
Vol 6 (3) ◽  
pp. e17833 ◽  
Author(s):  
Christian Franck ◽  
Stacey A. Maskarinec ◽  
David A. Tirrell ◽  
Guruswami Ravichandran
Keyword(s):  
Three Dimensional ◽  
Traction Force ◽  
Traction Force Microscopy ◽  
Force Microscopy ◽  
Cell Matrix ◽  
Matrix Interactions ◽  
Cell Matrix Interactions

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 Finite Element Analysis of Traction Force Microscopy: Influence of Cell Mechanics, Adhesion, and Morphology

2013 ◽  
Vol 135 (7) ◽  
Author(s):  
Rachel Zielinski ◽  
Cosmin Mihai ◽  
Douglas Kniss ◽  
Samir N. Ghadiali
Keyword(s):  
Sensitivity Analysis ◽  
Energy Density ◽  
Traction Force ◽  
Biomechanical Properties ◽  
Contractile Force ◽  
Adhesion Energy ◽  
Cell Stiffness ◽  
Adherent Cells ◽  
Force Microscopy ◽  
Contractile Forces

The interactions between adherent cells and their extracellular matrix (ECM) have been shown to play an important role in many biological processes, such as wound healing, morphogenesis, differentiation, and cell migration. Cells attach to the ECM at focal adhesion sites and transmit contractile forces to the substrate via cytoskeletal actin stress fibers. This contraction results in traction stresses within the substrate/ECM. Traction force microscopy (TFM) is an experimental technique used to quantify the contractile forces generated by adherent cells. In TFM, cells are seeded on a flexible substrate and displacements of the substrate caused by cell contraction are tracked and converted to a traction stress field. The magnitude of these traction stresses are normally used as a surrogate measure of internal cell contractile force or contractility. We hypothesize that in addition to contractile force, other biomechanical properties including cell stiffness, adhesion energy density, and cell morphology may affect the traction stresses measured by TFM. In this study, we developed finite element models of the 2D and 3D TFM techniques to investigate how changes in several biomechanical properties alter the traction stresses measured by TFM. We independently varied cell stiffness, cell-ECM adhesion energy density, cell aspect ratio, and contractility and performed a sensitivity analysis to determine which parameters significantly contribute to the measured maximum traction stress and net contractile moment. Results suggest that changes in cell stiffness and adhesion energy density can significantly alter measured tractions, independent of contractility. Based on a sensitivity analysis, we developed a correction factor to account for changes in cell stiffness and adhesion and successfully applied this correction factor algorithm to experimental TFM measurements in invasive and noninvasive cancer cells. Therefore, application of these types of corrections to TFM measurements can yield more accurate estimates of cell contractility.

scholarly journals Astigmatic traction force microscopy (aTFM)

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Di Li ◽  
Huw Colin-York ◽  
Liliana Barbieri ◽  
Yousef Javanmardi ◽  
Yuting Guo ◽  
...  
Keyword(s):  
Total Internal Reflection ◽  
Three Dimensional ◽  
Traction Force ◽  
Super Resolution ◽  
Traction Force Microscopy ◽  
Force Measurements ◽  
Complete Understanding ◽  
Force Microscopy ◽  
Cell Force ◽  
Limited Sensitivity

AbstractQuantifying small, rapidly progressing three-dimensional forces generated by cells remains a major challenge towards a more complete understanding of mechanobiology. Traction force microscopy is one of the most broadly applied force probing technologies but ascertaining three-dimensional information typically necessitates slow, multi-frame z-stack acquisition with limited sensitivity. Here, by performing traction force microscopy using fast single-frame astigmatic imaging coupled with total internal reflection fluorescence microscopy we improve the temporal resolution of three-dimensional mechanical force quantification up to 10-fold compared to its related super-resolution modalities. 2.5D astigmatic traction force microscopy (aTFM) thus enables live-cell force measurements approaching physiological sensitivity.

scholarly journals Publisher Correction: Epifluorescence-based three-dimensional traction force microscopy

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Lauren Hazlett ◽  
Alexander K. Landauer ◽  
Mohak Patel ◽  
Hadley A. Witt ◽  
Jin Yang ◽  
...  
Keyword(s):  
Three Dimensional ◽  
Traction Force ◽  
Traction Force Microscopy ◽  
Force Microscopy

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

scholarly journals Advanced in silico validation framework for three-dimensional Traction Force Microscopy and application to an in vitro model of sprouting angiogenesis

2020 ◽  
Author(s):  
J. Barrasa-Fano ◽  
A. Shapeti ◽  
J. de Jong ◽  
A. Ranga ◽  
J.A. Sanz-Herrera ◽  
...  
Keyword(s):  
In Silico ◽  
Inverse Method ◽  
In Vitro Model ◽  
Three Dimensional ◽  
Focal Adhesions ◽  
Traction Force ◽  
Force Microscopy ◽  
Validation Framework ◽  
Vitro Model

AbstractIn the last decade, cellular forces in three-dimensional hydrogels that mimic the extracellular matrix have been calculated by means of Traction Force Microscopy (TFM). However, characterizing the accuracy limits of a traction recovery method is critical to avoid obscuring physiological information due to traction recovery errors. So far, 3D TFM algorithms have only been validated using simplified cell geometries, bypassing image processing steps or arbitrarily simulating focal adhesions. Moreover, it is still uncertain which of the two common traction recovery methods, i.e., forward and inverse, is more robust against the inherent challenges of 3D TFM. In this work, we established an advanced in silico validation framework that is applicable to any 3D TFM experimental setup and that can be used to correctly couple the experimental and computational aspects of 3D TFM. Advancements relate to the simultaneous incorporation of complex cell geometries, simulation of microscopy images of varying bead densities and different focal adhesion sizes and distributions. By measuring the traction recovery error with respect to ground truth solutions, we found that while highest traction recovery errors occur for cases with sparse and small focal adhesions, our implementation of the inverse method improves two-fold the accuracy with respect to the forward method (average error of 23% vs. 50%). This advantage was further supported by recovering cellular tractions around angiogenic sprouts in an in vitro model of angiogenesis. The inverse method recovered more realistic traction patterns than the forward method, showing higher traction peaks and a clearer pulling pattern at the sprout protrusion tips.

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