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traction force microscopy

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https://www.readbyqxmd.com/read/28378710/calcium-oscillations-in-wounded-fibroblast-monolayers-are-spatially-regulated-through-substrate-mechanics
#1
Josephine Lembong, Benedikt Sabass, Howard Stone
The maintenance of tissue integrity is essential for the life of multicellular organisms. Healing of a skin wound is a paradigm for how various cell types localize and repair tissue perturbations in an orchestrated fashion. To investigate biophysical mechanisms associated with wound localization, we focus on a model system consisting of a fibroblast monolayer on an elastic substrate. We find that the creation of an edge in the monolayer causes cytosolic calcium oscillations throughout the monolayer. The oscillation frequency increases with cell density, which shows that wound-induced calcium oscillations occur collectively...
April 5, 2017: Physical Biology
https://www.readbyqxmd.com/read/28355484/podosome-force-generation-machinery-a-local-balance-between-protrusion-at-the-core-and-traction-at-the-ring
#2
Anaïs Bouissou, Amsha Proag, Nicolas Bourg, Karine Pingris, Clément Cabriel, Stéphanie Balor, Thomas Mangeat, Christophe Thibault, Christophe Vieu, Guillaume Dupuis, Emmanuel Fort, Sandrine Lévêque-Fort, Isabelle Maridonneau-Parini, Renaud Poincloux
Determining how cells generate and transduce mechanical forces at the nanoscale is a major technical challenge for the understanding of numerous physiological and pathological processes. Podosomes are submicrometer cell structures with a columnar F-actin core surrounded by a ring of adhesion proteins, which possess the singular ability to protrude into and probe the extracellular matrix. Using protrusion force microscopy, we have previously shown that single podosomes produce local nanoscale protrusions on the extracellular environment...
April 25, 2017: ACS Nano
https://www.readbyqxmd.com/read/28301462/dissection-of-mechanical-force-in-living-cells-by-super-resolved-traction-force-microscopy
#3
Huw Colin-York, Christian Eggeling, Marco Fritzsche
Cells continuously exert or respond to mechanical force. Measurement of these nanoscale forces is a major challenge in cell biology; yet such measurement is essential to the understanding of cell regulation and function. Current methods for examining mechanical force generation either necessitate dedicated equipment or limit themselves to coarse-grained force measurements on the micron scale. In this protocol, we describe stimulated emission depletion traction force microscopy-STED-TFM (STFM), which allows higher sampling of the forces generated by the cell than conventional TFM, leading to a twofold increase in spatial resolution (of up to 500 nm)...
April 2017: Nature Protocols
https://www.readbyqxmd.com/read/28287589/a-high-throughput-cell-microarray-platform-for-correlative-analysis-of-cell-differentiation-and-traction-forces
#4
Kerim B Kaylan, Andreas P Kourouklis, Gregory H Underhill
Microfabricated cellular microarrays, which consist of contact-printed combinations of biomolecules on an elastic hydrogel surface, provide a tightly controlled, high-throughput engineered system for measuring the impact of arrayed biochemical signals on cell differentiation. Recent efforts using cell microarrays have demonstrated their utility for combinatorial studies in which many microenvironmental factors are presented in parallel. However, these efforts have focused primarily on investigating the effects of biochemical cues on cell responses...
March 1, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/28271010/measurement-of-dynamic-cell-induced-3d-displacement-fields-in-vitro-for-traction-force-optical-coherence-microscopy
#5
Jeffrey A Mulligan, François Bordeleau, Cynthia A Reinhart-King, Steven G Adie
Traction force microscopy (TFM) is a method used to study the forces exerted by cells as they sense and interact with their environment. Cell forces play a role in processes that take place over a wide range of spatiotemporal scales, and so it is desirable that TFM makes use of imaging modalities that can effectively capture the dynamics associated with these processes. To date, confocal microscopy has been the imaging modality of choice to perform TFM in 3D settings, although multiple factors limit its spatiotemporal coverage...
February 1, 2017: Biomedical Optics Express
https://www.readbyqxmd.com/read/28256617/cholesterol-depletion-impairs-contractile-machinery-in-neonatal-rat-cardiomyocytes
#6
Barbara Hissa, Patrick W Oakes, Bruno Pontes, Guillermina Ramírez-San Juan, Margaret L Gardel
Cholesterol regulates numerous cellular processes. Depleting its synthesis in skeletal myofibers induces vacuolization and contraction impairment. However, little is known about how cholesterol reduction affects cardiomyocyte behavior. Here, we deplete cholesterol by incubating neonatal cardiomyocytes with methyl-beta-cyclodextrin. Traction force microscopy shows that lowering cholesterol increases the rate of cell contraction and generates defects in cell relaxation. Cholesterol depletion also increases membrane tension, Ca(2+) spikes frequency and intracellular Ca(2+) concentration...
March 3, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28255710/probing-synaptic-biomechanics-using-micropillar-arrays
#7
Weiyang Jin, Charles T Black, Lance C Kam, Morgan Huse
Recent insights into the importance of mechanosensing and force transmission at the immune synapse have spurred increased interest in the mechanical properties of leukocyte cell-cell interactions. In this chapter, we describe an imaging-based strategy for measuring cellular forces that utilizes optically transparent arrays of flexible micropillars. This approach has several distinct advantages over standard traction force microscopy, and we anticipate that it will prove very useful for investigators who wish not only to quantify ligand-induced forces with high spatiotemporal resolution but also to place those forces within the context of a broader cell biological response...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28164999/high-resolution-traction-force-microscopy-on-small-focal-adhesions-improved-accuracy-through-optimal-marker-distribution-and-optical-flow-tracking
#8
Claude N Holenstein, Unai Silvan, Jess G Snedeker
The accurate determination of cellular forces using Traction Force Microscopy at increasingly small focal attachments to the extracellular environment presents an important yet substantial technical challenge. In these measurements, uncertainty regarding accuracy is prominent since experimental calibration frameworks at this size scale are fraught with errors - denying a gold standard against which accuracy of TFM methods can be judged. Therefore, we have developed a simulation platform for generating synthetic traction images that can be used as a benchmark to quantify the influence of critical experimental parameters and the associated errors...
February 6, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28164414/microfluidic-traction-force-microscopy-to-study-mechanotransduction-in-angiogenesis
#9
Luke Boldock, Claudia Wittkowske, Cecile M Perrault
The formation of new blood vessels from existing vasculature, angiogenesis, is driven by coordinated endothelial cell migration and matrix remodelling in response to local signals. Recently, a growing body of evidence has shown that mechanotransduction, along with chemotransduction, is a major regulator of angiogenesis. Mechanical signals, such as fluid shear stress and substrate mechanics, influence sprouting and network formation, but the mechanisms behind this relationship are still unclear. Here, we present cellular traction forces as possible effectors activated by mechanosensing to mediate matrix remodelling, and encourage the use of traction force microscopy to study mechanotransduction in angiogenesis...
February 6, 2017: Microcirculation: the Official Journal of the Microcirculatory Society, Inc
https://www.readbyqxmd.com/read/28128382/reproducible-in-vitro-model-for-dystrophic-calcification-of-cardiac-valvular-interstitial-cells-insights-into-the-mechanisms-of-calcific-aortic-valvular-disease
#10
Heather A Cirka, Johana Uribe, Vivian Liang, Frederick J Schoen, Kristen L Billiar
Calcific aortic valvular disease (CAVD) is the most prevalent valvular pathology in the United States. Development of a pharmacologic agent to slow, halt, or reverse calcification has proven to be unsuccessful as still much remains unknown about the mechanisms of disease initiation. Although in vitro models of some features of CAVD exist, their utility is limited by the inconsistency of the size and time course of the calcified cell aggregates. In this study, we introduce and verify a highly reproducible in vitro method for studying dystrophic calcification of cardiac valvular interstitial cells, considered to be a key mechanism of clinical CAVD...
January 27, 2017: Lab on a Chip
https://www.readbyqxmd.com/read/28098306/effective-viscosity-and-dynamics-of-spreading-epithelia-a-solvable-model
#11
C Blanch-Mercader, R Vincent, E Bazellières, X Serra-Picamal, X Trepat, J Casademunt
Collective cell migration in spreading epithelia in controlled environments has become a landmark in our current understanding of fundamental biophysical processes in development, regeneration, wound healing or cancer. Epithelial monolayers are treated as thin layers of a viscous fluid that exert active traction forces on the substrate. The model is exactly solvable and shows a broad range of applicabilities for the quantitative analysis and interpretation of force microscopy data of monolayers from a variety of experiments and cell lines...
January 18, 2017: Soft Matter
https://www.readbyqxmd.com/read/28094390/simultaneous-measurement-of-the-young-s-modulus-and-the-poisson-ratio-of-thin-elastic-layers
#12
Wolfgang Gross, Holger Kress
The behavior of cells and tissue is greatly influenced by the mechanical properties of their environment. For studies on the interactions between cells and soft matrices, especially those applying traction force microscopy the characterization of the mechanical properties of thin substrate layers is essential. Various techniques to measure the elastic modulus are available. Methods to accurately measure the Poisson ratio of such substrates are rare and often imply either a combination of multiple techniques or additional equipment which is not needed for the actual biological studies...
February 7, 2017: Soft Matter
https://www.readbyqxmd.com/read/28074837/two-layer-elastographic-3-d-traction-force-microscopy
#13
Begoña Álvarez-González, Shun Zhang, Manuel Gómez-González, Ruedi Meili, Richard A Firtel, Juan C Lasheras, Juan C Del Álamo
Cellular traction force microscopy (TFM) requires knowledge of the mechanical properties of the substratum where the cells adhere to calculate cell-generated forces from measurements of substratum deformation. Polymer-based hydrogels are broadly used for TFM due to their linearly elastic behavior in the range of measured deformations. However, the calculated stresses, particularly their spatial patterns, can be highly sensitive to the substratum's Poisson's ratio. We present two-layer elastographic TFM (2LETFM), a method that allows for simultaneously measuring the Poisson's ratio of the substratum while also determining the cell-generated forces...
January 11, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28024085/on-the-functional-role-of-valve-interstitial-cell-stress-fibers-a-continuum-modeling-approach
#14
Yusuke Sakamoto, Rachel M Buchanan, Johannah Sanchez-Adams, Farshid Guilak, Michael S Sacks
The function of the heart valve interstitial cells (VICs) is intimately connected to heart valve tissue remodeling and repair, as well as the onset and progression of valvular pathological processes. There is yet only very limited knowledge and extant models for the complex three-dimensional VIC internal stress-bearing structures, the associated cell-level biomechanical behaviors, and how they change under varying activation levels. Importantly, VICs are known to exist and function within the highly dynamic valve tissue environment, including very high physiological loading rates...
February 1, 2017: Journal of Biomechanical Engineering
https://www.readbyqxmd.com/read/28008471/microfabricated-tissues-for-investigating-traction-forces-involved-in-cell-migration-and-tissue-morphogenesis
#15
REVIEW
Bryan A Nerger, Michael J Siedlik, Celeste M Nelson
Cell-generated forces drive an array of biological processes ranging from wound healing to tumor metastasis. Whereas experimental techniques such as traction force microscopy are capable of quantifying traction forces in multidimensional systems, the physical mechanisms by which these forces induce changes in tissue form remain to be elucidated. Understanding these mechanisms will ultimately require techniques that are capable of quantifying traction forces with high precision and accuracy in vivo or in systems that recapitulate in vivo conditions, such as microfabricated tissues and engineered substrata...
May 2017: Cellular and Molecular Life Sciences: CMLS
https://www.readbyqxmd.com/read/28002746/frustrated-phagocytic-spreading-of-j774a-1-macrophages-ends-in-myosin-ii-dependent-contraction
#16
Daniel T Kovari, Wenbin Wei, Patrick Chang, Jan-Simon Toro, Ruth Fogg Beach, Dwight Chambers, Karen Porter, Doyeon Koo, Jennifer E Curtis
Conventional studies of dynamic phagocytic behavior have been limited in terms of spatial and temporal resolution due to the inherent three-dimensionality and small features of phagocytosis. To overcome these issues, we use a series of frustrated phagocytosis assays to quantitatively characterize phagocytic spreading dynamics. Our investigation reveals that frustrated phagocytic spreading occurs in phases and is punctuated by a distinct period of contraction. The spreading duration and peak contact areas are independent of the surface opsonin density, although the opsonin density does affect the likelihood that a cell will spread...
December 20, 2016: Biophysical Journal
https://www.readbyqxmd.com/read/27994457/nanomechanical-measurement-of-adhesion-and-migration-of-leukemia-cells-with-phorbol-12-myristate-13-acetate-treatment
#17
Zhuo Long Zhou, Jing Ma, Ming-Hui Tong, Barbara Pui Chan, Alice Sze Tsai Wong, Alfonso Hing Wan Ngan
The adhesion and traction behavior of leukemia cells in their microenvironment is directly linked to their migration, which is a prime issue affecting the release of cancer cells from the bone marrow and hence metastasis. In assessing the effectiveness of phorbol 12-myristate 13-acetate (PMA) treatment, the conventional batch-cell transwell-migration assay may not indicate the intrinsic effect of the treatment on migration, since the treatment may also affect other cellular behavior, such as proliferation or death...
2016: International Journal of Nanomedicine
https://www.readbyqxmd.com/read/27990827/coordination-between-intra-and-extracellular-forces-regulates-focal-adhesion-dynamics
#18
Bibhu Ranjan Sarangi, Mukund Gupta, Bryant L Doss, Nicolas Tissot, France Lam, René-Marc Mège, Nicolas Borghi, Benoît Ladoux
Focal adhesions (FAs) are important mediators of cell-substrate interactions. One of their key functions is the transmission of forces between the intracellular acto-myosin network and the substrate. However, the relationships between cell traction forces, FA architecture, and molecular forces within FAs are poorly understood. Here, by combining Förster resonance energy transfer (FRET)-based molecular force biosensors with micropillar-based traction force sensors and high-resolution fluorescence microscopy, we simultaneously map molecular tension across vinculin, a key protein in FAs, and traction forces at FAs...
December 23, 2016: Nano Letters
https://www.readbyqxmd.com/read/27990663/%C3%AE-v%C3%AE-3-integrins-negatively-regulate-cellular-forces-by-phosphorylation-of-its-distal-npxy-site
#19
Rachel Milloud, Olivier Destaing, Richard de Mets, Ingrid Bourrin-Reynard, Christiane Oddou, Antoine Delon, Irène Wang, Corinne Albigès-Rizo, Martial Balland
BACKGROUND INFORMATION: Integrins are key receptors that allow cells to sense and respond to their mechanical environment. Although they bind the same ligand, β1 and β3 integrins have distinct and cooperative roles in mechanotransduction. RESULTS: Using traction force microscopy on unconstrained cells, we show that deleting β3 causes traction forces to increase, whereas the deletion of β1 integrin results in a strong decrease of contractile forces. Consistently, loss of β3 integrin also induces an increase in β1 integrin activation...
March 2017: Biology of the Cell
https://www.readbyqxmd.com/read/27892536/investigation-of-cell-substrate-adhesion-properties-of-living-chondrocyte-by-measuring-adhesive-shear-force-and-detachment-using-afm-and-inverse-fea
#20
Trung Dung Nguyen, YuanTong Gu
It is well-known that cell adhesion is important in many biological processes such as cell migration and proliferation. A better understanding of the cell adhesion process will shed insight into these cellular biological responses as well as cell adhesion-related diseases treatment. However, there is little research which has attempted to investigate the process of cell adhesion and its mechanism. Thus, this paper aims to study the time-dependent adhesion properties of single living chondrocytes using an advanced coupled experimental-numerical approach...
November 28, 2016: Scientific Reports
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