Generation of biocompatible droplets for in vivo and in vitro measurement of cell-generated mechanical stresses

Lucio, Adam; Ingber, Donald E.; Campàs, Otger

doi:10.1016/bs.mcb.2014.11.008

Cited by 16 publications

(19 citation statements)

References 34 publications

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“…Droplets were generated and coated as described in Lucio et al . () and mixed with an aqueous solution containing low melting point agarose at 1% (v/v). Temperature was lowered below the gelation point, immobilizing the droplets in the agarose gel.…”

Section: Analysis Of Experimental Datamentioning

confidence: 99%

“…After having characterized the performance of the algorithm with controlled, synthetic data, we tested its performance with experimental data. To do so, we first used fluorocarbon oil droplets with fluorescently labelled surfaces, as previously described (Campàs et al, 2014;Lucio et al, 2015). Droplets were generated and coated as described in Lucio et al (2015) and mixed with an aqueous solution containing low melting point agarose at 1% (v/v).…”

Section: Analysis Of Experimental Datamentioning

confidence: 99%

“…To do so, we first used fluorocarbon oil droplets with fluorescently labelled surfaces, as previously described (Campàs et al, 2014;Lucio et al, 2015). Droplets were generated and coated as described in Lucio et al (2015) and mixed with an aqueous solution containing low melting point agarose at 1% (v/v). Temperature was lowered below the gelation point, immobilizing the droplets in the agarose gel.…”

Section: Analysis Of Experimental Datamentioning

confidence: 99%

“…After having established that the developed algorithm is able to provide accurate segmentation and curvature maps of surfaces, we use it to characterize the geometry of oil droplets deformed by cellular forces. Fluorocarbon oil droplets similar in size to individual cells, coated with ligands (RGD) targeting integrin receptors, were generated as recently reported (Campàs et al, 2014;Lucio et al, 2015). These droplets were injected in multicellular aggregates of tooth mesenchymal cells and subsequently imaged using previously described protocols (Campàs et al, 2014).…”

Section: Analysis Of Experimental Datamentioning

confidence: 99%

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Geometrical characterization of fluorescently labelled surfaces from noisy 3D microscopy data

Shelton

Serwane

Campàs

2017

Journal of Microscopy

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Modern fluorescence microscopy enables fast 3D imaging of biological and inert systems alike. In many studies, it is important to detect the surface of objects and quantitatively characterize its local geometry, including its mean curvature. We present a fully automated algorithm to determine the location and curvatures of an object from 3D fluorescence images, such as those obtained using confocal or light-sheet microscopy. The algorithm aims at reconstructing surface labelled objects with spherical topology and mild deformations from the spherical geometry with high accuracy, rather than reconstructing arbitrarily deformed objects with lower fidelity. Using both synthetic data with known geometrical characteristics and experimental data of spherical objects, we characterize the algorithm's accuracy over the range of conditions and parameters typically encountered in 3D fluorescence imaging. We show that the algorithm can detect the location of the surface and obtain a map of local mean curvatures with relative errors typically below 2% and 20%, respectively, even in the presence of substantial levels of noise. Finally, we apply this algorithm to analyse the shape and curvature map of fluorescently labelled oil droplets embedded within multicellular aggregates and deformed by cellular forces.

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Section: Analysis Of Experimental Datamentioning

confidence: 99%

Section: Analysis Of Experimental Datamentioning

confidence: 99%

Section: Analysis Of Experimental Datamentioning

confidence: 99%

Section: Analysis Of Experimental Datamentioning

confidence: 99%

See 2 more Smart Citations

Geometrical characterization of fluorescently labelled surfaces from noisy 3D microscopy data

Shelton

Serwane

Campàs

2017

Journal of Microscopy

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“…Initially developed to measure the mechanical stresses generated by growing actin networks in vitro [225,226], this technique has recently been adapted to measure mechanical stresses within living embryonic tissues [227,228]. DS use cell-sized, biocompatible oil microdroplets, coated with ligands for cell adhesion receptors, as force transducers in vivo and in situ (and also ex vivo and in vitro ).…”

Section: Methodologies To Probe the Mechanics Of Living Embryonic mentioning

confidence: 99%

A toolbox to explore the mechanics of living embryonic tissues

Campàs

2016

Seminars in Cell & Developmental Biology

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128

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The sculpting of embryonic tissues and organs into their functional morphologies involves the spatial and temporal regulation of mechanics at cell and tissue scales. Decades of in vitro work, complemented by some in vivo studies, have shown the relevance of mechanical cues in the control of cell behaviors that are central to developmental processes, but the lack of methodologies enabling precise, quantitative measurements of mechanical cues in vivo have hindered our understanding of the role of mechanics in embryonic development. Several methodologies are starting to enable quantitative studies of mechanics in vivo and in situ, opening new avenues to explore how mechanics contributes to shaping embryonic tissues and how it affects cell behavior within developing embryos. Here we review the present methodologies to study the role of mechanics in living embryonic tissues, considering their strengths and drawbacks as well as the conditions in which they are most suitable.

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Generation, Transmission, and Regulation of Mechanical Forces in Embryonic Morphogenesis

Sutlive

Xiu

Chen

et al. 2021

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Embryonic morphogenesis is a biological process which depicts shape forming of tissues and organs during development. Unveiling the roles of mechanical forces generated, transmitted, and regulated in cells and tissues through these processes is key to understanding the biophysical mechanisms governing morphogenesis. To this end, it is imperative to measure, simulate, and predict the regulation and control of these mechanical forces during morphogenesis. This article aims to provide a comprehensive review of the recent advances on mechanical properties of cells and tissues, generation of mechanical forces in cells and tissues, the transmission processes of these generated forces during cells and tissues, the tools and methods used to measure and predict these mechanical forces in vivo, in vitro, or in silico, and to better understand the corresponding regulation and control of generated forces. Understanding the biomechanics and mechanobiology of morphogenesis will not only shed light on the fundamental physical mechanisms underlying these concerted biological processes during normal development, but also uncover new information that will benefit biomedical research in preventing and treating congenital defects or tissue engineering and regeneration.

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Generation of biocompatible droplets for in vivo and in vitro measurement of cell-generated mechanical stresses

Cited by 16 publications

References 34 publications

Geometrical characterization of fluorescently labelled surfaces from noisy 3D microscopy data

Geometrical characterization of fluorescently labelled surfaces from noisy 3D microscopy data

A toolbox to explore the mechanics of living embryonic tissues

Generation, Transmission, and Regulation of Mechanical Forces in Embryonic Morphogenesis

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