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traction force microscopy

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https://www.readbyqxmd.com/read/28797233/full-l1-regularized-traction-force-microscopy-over-whole-cells
#1
Alejandro Suñé-Auñón, Alvaro Jorge-Peñas, Rocío Aguilar-Cuenca, Miguel Vicente-Manzanares, Hans Van Oosterwyck, Arrate Muñoz-Barrutia
BACKGROUND: Traction Force Microscopy (TFM) is a widespread technique to estimate the tractions that cells exert on the surrounding substrate. To recover the tractions, it is necessary to solve an inverse problem, which is ill-posed and needs regularization to make the solution stable. The typical regularization scheme is given by the minimization of a cost functional, which is divided in two terms: the error present in the data or data fidelity term; and the regularization or penalty term...
August 10, 2017: BMC Bioinformatics
https://www.readbyqxmd.com/read/28793219/stiff-substrates-increase-inflammation-induced-endothelial-monolayer-tension-and-permeability
#2
Rebecca Lownes Urbano, Christina Furia, Sarah Basehore, Alisa Morss Clyne
Arterial stiffness and inflammation are associated with atherosclerosis, and each have individually been shown to increase endothelial monolayer tension and permeability. The objective of this study was to determine if substrate stiffness enhanced endothelial monolayer tension and permeability in response to inflammatory cytokines. Porcine aortic endothelial cells were cultured at confluence on polyacrylamide gels of varying stiffness and treated with either tumor necrosis factor-α (TNFα) or thrombin. Monolayer tension was measured through vinculin localization at the cell membrane, traction force microscopy, and phosphorylated myosin light chain quantity and actin fiber colocalization...
August 8, 2017: Biophysical Journal
https://www.readbyqxmd.com/read/28753694/endogenous-sheet-averaged-tension-within-a-large-epithelial-cell-colony
#3
Sandeep P Dumbali, Lanju Mei, Shizhi Qian, Venkat Maruthamuthu
Epithelial cells form quasi-two-dimensional sheets that function as contractile media to effect tissue shape changes during development and homeostasis. Endogenously generated intrasheet tension is a driver of such changes, but has predominantly been measured in the presence of directional migration. The nature of epithelial cell-generated forces transmitted over supracellular distances, in the absence of directional migration, is thus largely unclear. In this report, we consider large epithelial cell colonies which are archetypical multicell collectives with extensive cell-cell contacts but with a symmetric (circular) boundary...
October 1, 2017: Journal of Biomechanical Engineering
https://www.readbyqxmd.com/read/28726917/engineering-micromyocardium-to-delineate-cellular-and-extracellular-regulation-of-myocardial-tissue-contractility
#4
Nethika R Ariyasinghe, Caitlin H Reck, Alyssa A Viscio, Andrew P Petersen, Davi M Lyra-Leite, Nathan Cho, Megan L McCain
Cardiovascular diseases are a leading cause of death, in part due to limitations of existing models of the myocardium. Myocardium consists of aligned, contractile cardiac myocytes interspersed with fibroblasts that synthesize extracellular matrix (ECM). The cellular demographics and biochemical and mechanical properties of the ECM remodel in many different cardiac diseases. However, the impact of diverse cellular and extracellular remodeling on the contractile output of the myocardium are poorly understood...
July 20, 2017: Integrative Biology: Quantitative Biosciences From Nano to Macro
https://www.readbyqxmd.com/read/28708434/epithelial-cells-induce-a-cyclooxygenase-1-dependent-endogenous-reduction-in-airway-smooth-muscle-contractile-phenotype
#5
Michael J O'Sullivan, Elizabeth Gabriel, Alice Panariti, Chan Y Park, Gijs Ijpma, Jeffrey J Fredberg, Anne-Marie Lauzon, James G Martin
Airway smooth muscle (ASM) cells are phenotypically regulated to exist in either a proliferative or a contractile state. However the influence of other airway structural cell types on ASM cell phenotype is largely unknown. Although epithelial cells are known to drive ASM proliferation, their effects on the contractile phenotype are uncertain. In the current study we tested the hypothesis that epithelial cells reduce the contractile phenotype of ASM cells. To do so, we measured force production by traction microscopy, gene and protein expression as well as calcium release by Fura-2 ratiometric imaging...
July 14, 2017: American Journal of Respiratory Cell and Molecular Biology
https://www.readbyqxmd.com/read/28674019/changes-in-e-cadherin-rigidity-sensing-regulate-cell-adhesion
#6
Caitlin Collins, Aleksandra K Denisin, Beth L Pruitt, W James Nelson
Mechanical cues are sensed and transduced by cell adhesion complexes to regulate diverse cell behaviors. Extracellular matrix (ECM) rigidity sensing by integrin adhesions has been well studied, but rigidity sensing by cadherins during cell adhesion is largely unexplored. Using mechanically tunable polyacrylamide (PA) gels functionalized with the extracellular domain of E-cadherin (Ecad-Fc), we showed that E-cadherin-dependent epithelial cell adhesion was sensitive to changes in PA gel elastic modulus that produced striking differences in cell morphology, actin organization, and membrane dynamics...
July 18, 2017: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/28655845/force-generation-by-groups-of-migrating-bacteria
#7
Benedikt Sabass, Matthias D Koch, Guannan Liu, Howard A Stone, Joshua W Shaevitz
From colony formation in bacteria to wound healing and embryonic development in multicellular organisms, groups of living cells must often move collectively. Although considerable study has probed the biophysical mechanisms of how eukaryotic cells generate forces during migration, little such study has been devoted to bacteria, in particular with regard to the question of how bacteria generate and coordinate forces during collective motion. This question is addressed here using traction force microscopy. We study two distinct motility mechanisms of Myxococcus xanthus, namely, twitching and gliding...
July 11, 2017: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/28633918/traction-force-and-its-regulation-during-cytokinesis-in-dictyostelium-cells
#8
Md Golam Sarowar Jahan, Shigehiko Yumura
Cytokinesis is the final stage of cell division. Dictyostelium cells have multiple modes of cytokinesis, including cytokinesis A, B and C. Cytokinesis A is a conventional mode, which depends on myosin II in the contractile ring. Myosin II null cells divide depending on substratum-attachment (cytokinesis B) or in a multi-polar fashion independent of the cell cycle (cytokinesis C). We investigated the traction stress exerted by dividing cells in the three different modes using traction force microscopy. In all cases, the traction forces were directed inward from both poles...
June 12, 2017: European Journal of Cell Biology
https://www.readbyqxmd.com/read/28627753/traction-force-microscopy-in-3-dimensional-extracellular-matrix-networks
#9
M Cóndor, J Steinwachs, C Mark, J M García-Aznar, B Fabry
Cell migration through a three-dimensional (3-D) matrix depends strongly on the ability of cells to generate traction forces. To overcome the steric hindrance of the matrix, cells need to generate sufficiently high traction forces but also need to distribute these forces spatially in a migration-promoting way. This unit describes a protocol to measure spatial maps of cell traction forces in 3-D biopolymer networks such as collagen, fibrin, or Matrigel. Traction forces are computed from the relationship between measured force-induced matrix deformations surrounding the cell and the known mechanical properties of the matrix...
June 19, 2017: Current Protocols in Cell Biology
https://www.readbyqxmd.com/read/28604737/optogenetic-control-of-rhoa-reveals-zyxin-mediated-elasticity-of-stress-fibres
#10
Patrick W Oakes, Elizabeth Wagner, Christoph A Brand, Dimitri Probst, Marco Linke, Ulrich S Schwarz, Michael Glotzer, Margaret L Gardel
Cytoskeletal mechanics regulates cell morphodynamics and many physiological processes. While contractility is known to be largely RhoA-dependent, the process by which localized biochemical signals are translated into cell-level responses is poorly understood. Here we combine optogenetic control of RhoA, live-cell imaging and traction force microscopy to investigate the dynamics of actomyosin-based force generation. Local activation of RhoA not only stimulates local recruitment of actin and myosin but also increased traction forces that rapidly propagate across the cell via stress fibres and drive increased actin flow...
June 12, 2017: Nature Communications
https://www.readbyqxmd.com/read/28532442/an-integrated-enhancement-and-reconstruction-strategy-for-the-quantitative-extraction-of-actin-stress-fibers-from-fluorescence-micrographs
#11
Zhen Zhang, Shumin Xia, Pakorn Kanchanawong
BACKGROUND: The stress fibers are prominent organization of actin filaments that perform important functions in cellular processes such as migration, polarization, and traction force generation, and whose collective organization reflects the physiological and mechanical activities of the cells. Easily visualized by fluorescence microscopy, the stress fibers are widely used as qualitative descriptors of cell phenotypes. However, due to the complexity of the stress fibers and the presence of other actin-containing cellular features, images of stress fibers are relatively challenging to quantitatively analyze using previously developed approaches, requiring significant user intervention...
May 22, 2017: BMC Bioinformatics
https://www.readbyqxmd.com/read/28521203/3d-full-field-quantification-of-cell-induced-large-deformations-in-fibrillar-biomaterials-by-combining-non-rigid-image-registration-with-label-free-second-harmonic-generation
#12
Alvaro Jorge-Peñas, Hannelore Bové, Kathleen Sanen, Marie-Mo Vaeyens, Christian Steuwe, Maarten Roeffaers, Marcel Ameloot, Hans Van Oosterwyck
To advance our current understanding of cell-matrix mechanics and its importance for biomaterials development, advanced three-dimensional (3D) measurement techniques are necessary. Cell-induced deformations of the surrounding matrix are commonly derived from the displacement of embedded fiducial markers, as part of traction force microscopy (TFM) procedures. However, these fluorescent markers may alter the mechanical properties of the matrix or can be taken up by the embedded cells, and therefore influence cellular behavior and fate...
May 10, 2017: Biomaterials
https://www.readbyqxmd.com/read/28476967/in-situ-fabrication-of-zirconium-titanium-nano-composite-and-its-coating-on-ti-6al-4v-for-biomedical-applications
#13
Manickam Chellappa, Uthirapathy Vijayalakshmi
In this study, nanocomposite powder consisting of zirconia and titania (Zr-Ti) have been synthesised by sol-gel method, with the aim of protecting Ti-6Al-4V surface. A simple and low cost electrophoretic deposition (EPD) technique has been employed for coating the nanocomposite material on Ti-6Al-4V. The prepared nanocomposite powder was characterised for its functional groups, phase purity, surface topography by Fourier transform infrared spectroscopy, powder X-ray diffraction and scanning electron microscopy...
February 2017: IET Nanobiotechnology
https://www.readbyqxmd.com/read/28458726/effect-of-m1-m2-polarization-on-the-motility-and-traction-stresses-of-primary-human-macrophages
#14
Laurel E Hind, Emily B Lurier, Micah Dembo, Kara L Spiller, Daniel A Hammer
Macrophages become polarized by cues in their environment and this polarization causes a functional change in their behavior. Two main subsets of polarized macrophages have been described. M1, or "classically activated" macrophages, are pro-inflammatory and M2, or "alternatively activated" macrophages, are anti-inflammatory. In this study, we investigated the motility and force generation of primary human macrophages polarized down the M1 and M2 pathways using chemokinesis assays and traction force microscopy on polyacrylamide gels...
September 2016: Cellular and Molecular Bioengineering
https://www.readbyqxmd.com/read/28378710/calcium-oscillations-in-wounded-fibroblast-monolayers-are-spatially-regulated-through-substrate-mechanics
#15
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
#16
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
#17
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
#18
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
#19
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
#20
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
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