Long-term Brillouin imaging of live cells with reduced absorption-mediated damage at 660nm wavelength

Nikolić, Miloš; Scarcelli, Giuliano

doi:10.1101/535419

Cited by 2 publications

(3 citation statements)

References 33 publications

(31 reference statements)

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“…To ensure that mechanical heterogeneities were not driving cell migration to the microwell edge via durotaxis (41,42), we mapped the local longitudinal moduli of the hydrogels using Brillouin microscopy (43,44). The Brillouin frequency shift is sensitive to changes in material stiffness and has previously been used to characterize several types of soft materials, including gelatin-based hydrogels (45).…”

Section: The Topography and Mechanical Properties Of The Hydrogel Microwells Are Uniformmentioning

confidence: 99%

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Directing multicellular organization by varying the aspect ratio of soft hydrogel microwells

Pahapale¹,

Tao

Nikolić

et al. 2021

Preprint

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Multicellular organization with precise spatial definition is an essential step in a wide range of biological processes, including morphogenesis, development, and healing. Gradients and patterns of chemoattractants are well-described guides of multicellular organization, but the influences of three-dimensional geometry of soft hydrogels on multicellular organization are less well defined. Here, we report the discovery of a new mode of self-organization of endothelial cells in ring-like patterns on the perimeters of hydrogel microwells that is independent of protein or chemical patterning and is driven only by geometry and substrate stiffness. We observe quantitatively striking influences of both the microwell aspect ratio (ε = perimeter/depth) and the hydrogel modulus. We systematically investigate the physical factors of cells and substrates that drive this multicellular behavior and present a mathematical model that explains the multicellular organization based upon balancing extracellular and cytoskeletal forces. These forces are determined in part by substrate stiffness, geometry, and cell density. The force balance model predicts the direction and distance of translational cell migration based on the dynamic interaction between tangential cytoskeletal tension and cell-cell and cell-substrate adhesion. We further show that the experimental observations can be leveraged to drive customized multicellular self-organization. Our observation of this multicellular behavior demonstrates the importance of the combinatorial effects of geometry and stiffness in complex biological processes. It also provides a new methodology for direction of cell organization that may facilitate the engineering of bionics and integrated model organoid systems.

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Section: The Topography and Mechanical Properties Of The Hydrogel Microwells Are Uniformmentioning

confidence: 99%

“…We measured the Brillouin shift of the hydrogels using a twostage Virtually Imaged Phase Array (VIPA) spectrometer as described previously (44,67). In brief, the sample was illuminated with 660 nm laser light through a 60x, 0.7NA objective (Olympus).…”

Section: Brillouin Microscopy Measurementsmentioning

confidence: 99%

Directing multicellular organization by varying the aspect ratio of soft hydrogel microwells

Pahapale¹,

Tao

Nikolić

et al. 2021

Preprint

Self Cite

View full text Add to dashboard Cite

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“…However, it also decreases the signal intensity due to the inverse relationship with the forth power of incident wavelength (as well as reducing the frequency shift). Nikolic and Scarcelli [8] showed that the use of 660 nm wavelength enables live cells to be irradiated with 82 times more energy than at 532 nm, thus shortening the acquisition time (by ca. 34 times).…”

Section: Introductionmentioning

confidence: 99%

Brillouin Light Scattering Microspectroscopy for Biomedical Research and Applications: introduction to feature issue

Elsayad¹,

Palombo²,

Dehoux³

et al. 2019

Biomed. Opt. Express

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There has been a marked revival of interest in brillouin light scattering spectroscopy/microscopy over the last decade in regards to applications related to all optically studying the mechanical problems associated with systems of biological and medical interest. This revival has been driven by advancements in spectrometer design, together with mounting evidence of the critical role that mechanical properties can play in biological processes as well as the onset of diverse diseases. This feature issue contains a series of papers spanning some of the latest developments in the field of Brillouin light scattering spectroscopy and microscopy as applied to systems of biomedical interest.

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Long-term Brillouin imaging of live cells with reduced absorption-mediated damage at 660nm wavelength

Cited by 2 publications

References 33 publications

Directing multicellular organization by varying the aspect ratio of soft hydrogel microwells

Directing multicellular organization by varying the aspect ratio of soft hydrogel microwells

Brillouin Light Scattering Microspectroscopy for Biomedical Research and Applications: introduction to feature issue

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