Spatiotemporal three-dimensional transport dynamics of endocytic cargos and their physical regulations in cells

Jiang, Chao; Yang, Mingcheng; Li, Wei; Dou, S. X.; Wang, Peng‐Ye; Li, Hui

doi:10.1016/j.isci.2022.104210

Cited by 13 publications

(20 citation statements)

References 71 publications

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“… (F) Fraction of confined (a < 0.5), subdiffusive (0.5<a < 1), and superdiffusive (1<a < 2) motions in the two different regions. Figure (D–F) is reproduced from Jiang et al. (2022) .…”

Section: Step-by-step Methods Detailsmentioning

confidence: 99%

“…This protocol can be used to characterize the intracellular diffusion and trafficking of macromolecules, nanoparticles, and endocytic vesicles in adherent cells. For complete details on the use and execution of this protocol, please refer to Jiang et al. (2022) .…”

mentioning

confidence: 99%

“…For complete details on the use and execution of this protocol, please refer to Jiang et al. (2022) .…”

mentioning

confidence: 99%

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Quantifying transport dynamics with three-dimensional single-particle tracking in adherent cells

Jiang¹,

Dou²,

Wang³

et al. 2022

STAR Protocols

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“… (F) Fraction of confined (a < 0.5), subdiffusive (0.5<a < 1), and superdiffusive (1<a < 2) motions in the two different regions. Figure (D–F) is reproduced from Jiang et al. (2022) .…”

Section: Step-by-step Methods Detailsmentioning

confidence: 99%

mentioning

confidence: 99%

See 1 more Smart Citation

Quantifying transport dynamics with three-dimensional single-particle tracking in adherent cells

Jiang¹,

Dou²,

Wang³

et al. 2022

STAR Protocols

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“…Deciphering the kinetics, transport, and localization of individual molecules is specifically important as it often uncovers mechanisms veiled in ensemble experiments . Fluorescence microscopy (FM) comes with the advantage of noninvasive imaging, and higher specificities have aided numerous cellular and molecular studies, including investigations of EVs. , 3D single-molecule tracking helped to reveal intercellular transport of transferrin; it also was used to investigate spatiotemporal transport dynamics of endocytic nanoparticles in cells . Two-color quantum-dot tracking was used for quantification of receptor dimerization rates .…”

Section: Introductionmentioning

confidence: 99%

“… 17 , 24 3D single-molecule tracking helped to reveal intercellular transport of transferrin; 25 it also was used to investigate spatiotemporal transport dynamics of endocytic nanoparticles in cells. 25 Two-color quantum-dot tracking was used for quantification of receptor dimerization rates. 26 There have been also numerous methodological advances in the study of EVs, particularly in relation to their uptake and internalization.…”

Section: Introductionmentioning

confidence: 99%

Purification Analysis, Intracellular Tracking, and Colocalization of Extracellular Vesicles Using Atomic Force and 3D Single-Molecule Localization Microscopy

et al. 2023

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Extracellular vesicles (EVs) play a key role in cell− cell communication and thus have great potential to be utilized as therapeutic agents and diagnostic tools. In this study, we implemented single-molecule microscopy techniques as a toolbox for a comprehensive characterization as well as measurement of the cellular uptake of HEK293T cell-derived EVs (eGFP-labeled) in HeLa cells. A combination of fluorescence and atomic force microscopy revealed a fraction of 68% fluorescently labeled EVs with an average size of ∼45 nm. Two-color single-molecule fluorescence microscopy analysis elucidated the 3D dynamics of EVs entering HeLa cells. 3D colocalization analysis of two-color direct stochastic optical reconstruction microscopy (dSTORM) images revealed that 25% of EVs that experienced uptake colocalized with transferrin, which has been linked to early recycling of endosomes and clathrin-mediated endocytosis. The localization analysis was combined with stepwise photobleaching, providing a comparison of protein aggregation outside and inside the cells.

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Interplay Between Intracellular Transport Dynamics and Liquid‒Liquid Phase Separation

Zhang,

Niu

et al. 2024

Advanced Science

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Liquid‒liquid phase separation (LLPS) is a ubiquitous process in which proteins, RNA, and biomolecules assemble into membrane‐less compartments, playing important roles in many biological functions and diseases. The current knowledge on the biophysical and biochemical principles of LLPS is largely from in vitro studies; however, the physiological environment in living cells is complex and not at equilibrium. The characteristics of intracellular dynamics and their roles in physiological LLPS remain to be resolved. Here, by using single‐particle tracking of quantum dots and dynamic monitoring of the formation of stress granules (SGs) in single cells, the spatiotemporal dynamics of intracellular transport in cells undergoing LLPS are quantified. It is shown that intracellular diffusion and active transport are both reduced. Furthermore, the formation of SG droplets contributes to increased spatial heterogeneity within the cell. More importantly, the study demonstrated that the LLPS of SGs can be regulated by intracellular dynamics in two stages: the reduced intracellular diffusion promotes SG assembly and the microtubule‐associated transport facilitates SG coalescences. The work on intracellular dynamics not only improves the understanding of the mechanism of physiology phase separations occurring in nonequilibrium environments but also reveals an interplay between intracellular dynamics and LLPS.

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Spatiotemporal three-dimensional transport dynamics of endocytic cargos and their physical regulations in cells

Cited by 13 publications

References 71 publications

Quantifying transport dynamics with three-dimensional single-particle tracking in adherent cells

Quantifying transport dynamics with three-dimensional single-particle tracking in adherent cells

Purification Analysis, Intracellular Tracking, and Colocalization of Extracellular Vesicles Using Atomic Force and 3D Single-Molecule Localization Microscopy

Interplay Between Intracellular Transport Dynamics and Liquid‒Liquid Phase Separation

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