Molecular force sensors to measure stress in cells

Prabhune, Meenakshi; Rehfeldt, Florian; Schmidt, Christoph F.

doi:10.1088/1361-6463/aa6e1e

Cited by 17 publications

(15 citation statements)

References 126 publications

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“…This offers the opportunity to use such functionalized beads as probes to quantify mechanical stress acting on them at the cellular scale. While molecular scale stress sensors are now frequently used, most commonly in the form of FRET dyes attached to calibrated molecules that unfold when being part of a molecular force chain, cell-scale stress sensors are still not used widely (89). Sensors based on oil droplets have been used to measure stresses in tissue, but with such incompressible liquids isotropic and shear certified by peer review) is the author/funder.…”

Section: Beads As Passive Cell-scale Stress Sensorsmentioning

confidence: 99%

Standardized microgel beads as elastic cell mechanical probes

et al. 2018

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Section: Beads As Passive Cell-scale Stress Sensorsmentioning

confidence: 99%

Standardized microgel beads as elastic cell mechanical probes

et al. 2018

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Section: Beads As Passive Cell-scale Stress Sensorsmentioning

confidence: 99%

Standardized microgel beads as elastic cell mechanical probes

Girardo

Traeber

Wagner

et al. 2018

Preprint

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Cell mechanical measurements are gaining increasing interest in biological and biomedical studies. However, there are no standardized calibration particles available that permit the cross-comparison of different measurement techniques operating at different stresses and time-scales. Here we present the rational design, production, and comprehensive characterization of poly-acylamide (PAAm) microgel beads mimicking biological cells. We produced mono-disperse beads at rates of 20 -60 kHz by means of a microfluidic droplet generator, where the pre-gel composition was adjusted to tune the beads' elasticity in the range of cell and tissue relevant mechanical properties. We verified bead homogeneity by optical diffraction tomography and Brillouin microscopy. Consistent elastic behavior of microgel beads at different shear rates was confirmed by AFM-enabled nanoindentation and real-time deformability cytometry (RT-DC). The remaining inherent variability in elastic modulus was rationalized using polymer theory and effectively reduced by sorting based on forward-scattering using conventional flow cytometry. Our results show that PAAm microgel beads can be standardized as mechanical probes, to serve not only for validation and calibration of cell mechanical measurements, but also as cell-scale stress sensors. Significance StatementOften vastly different cell mechanical properties are reported even for the same cell type when employing different measurement techniques. This discrepancy shows the urgent need for standardized calibration particles to cross-compare and validate techniques. Microgel beads can serve this purpose, but they have to fulfil specific requirements such as homogeneity, sizes and elasticities in the range of the cells, and they have to provide comparable results independent of the method applied. Here we demonstrate the standardized production of polyacrylamide microgel beads with all the features an elastic cell-mimic should have. These can not only be used as method calibration particles, but can also serve as cell-scale sensors to quantify normal and shear stresses exerted by other cells and inside tissues, enabling many new applications.

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“…We do not detail the latest technical developments in the field of intracellular mechanics and refer to Refs. [6][7][8][9][10][11][12] for recent reviews on the technical aspects.…”

Section: Introductionmentioning

confidence: 99%

Intracellular mechanics: connecting rheology and mechanotransduction

Mathieu

Manneville

2019

Current Opinion in Cell Biology

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Cell mechanics is crucial for a wide range of cell functions, including proliferation, polarity, migration and differentiation. Cells sense external physical cues and translate them into a cellular response. While force sensing occurs in the vicinity of the plasma membrane, forces can reach deep in the cell interior and to the nucleus. We review here the recent developments in the field of intracellular mechanics. We focus first on intracellular rheology, the study of the mechanical properties of the cell interior, and recapitulate the contribution of active mechanisms, the cytoskeleton and intracellular organelles to cell rheology. We then discuss how forces are transmitted inside the cell during mechanotransduction events, through direct force transmission and biochemical signaling, and how intracellular rheology and mechanotransduction are connected.

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Molecular force sensors to measure stress in cells

Cited by 17 publications

References 126 publications

Standardized microgel beads as elastic cell mechanical probes

Standardized microgel beads as elastic cell mechanical probes

Standardized microgel beads as elastic cell mechanical probes

Intracellular mechanics: connecting rheology and mechanotransduction

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