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3D tracking of extracellular vesicles by holographic fluorescence imaging.


ABSTRACT: Fluorescence microscopy is the method of choice in biology for its molecular specificity and super-resolution capabilities. However, it is limited to a narrow z range around one observation plane. Here, we report an imaging approach that recovers the full electric field of fluorescent light with single-molecule sensitivity. We expand the principle of digital holography to fast fluorescent detection by eliminating the need for phase cycling and enable three-dimensional (3D) tracking of individual nanoparticles with an in-plane resolution of 15 nm and a z-range of 8 mm. As a proof-of-concept biological application, we image the 3D motion of extracellular vesicles (EVs) inside live cells. At short time scales (<4 s), we resolve near-isotropic 3D diffusion and directional transport. For longer lag times, we observe a transition toward anisotropic motion with the EVs being transported over long distances in the axial plane while being confined in the horizontal dimension.

SUBMITTER: Liebel M 

PROVIDER: S-EPMC7673696 | biostudies-literature | 2020 Nov

REPOSITORIES: biostudies-literature

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3D tracking of extracellular vesicles by holographic fluorescence imaging.

Liebel Matz M   Ortega Arroyo Jaime J   Beltrán Vanesa Sanz VS   Osmond Johann J   Jo Ala A   Lee Hakho H   Quidant Romain R   van Hulst Niek F NF  

Science advances 20201104 45


Fluorescence microscopy is the method of choice in biology for its molecular specificity and super-resolution capabilities. However, it is limited to a narrow <i>z</i> range around one observation plane. Here, we report an imaging approach that recovers the full electric field of fluorescent light with single-molecule sensitivity. We expand the principle of digital holography to fast fluorescent detection by eliminating the need for phase cycling and enable three-dimensional (3D) tracking of ind  ...[more]

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