3D super-resolution deep-tissue imaging in living mice

Stimulated emission depletion (STED) microscopy enables the three-dimensional (3D) visualization of dynamic nanoscale structures in living cells, offering unique insights into their organization. However, 3D-STED imaging deep inside biological tissue is obstructed by optical aberrations and light sc...

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Published inOptica Vol. 8; no. 4; pp. 442 - 450
Main Authors Velasco, Mary Grace M, Zhang, Mengyang, Antonello, Jacopo, Yuan, Peng, Allgeyer, Edward S, May, Dennis, M'Saad, Ons, Kidd, Phylicia, Barentine, Andrew E S, Greco, Valentina, Grutzendler, Jaime, Booth, Martin J, Bewersdorf, Joerg
Format Journal Article
LanguageEnglish
Published United States Optical Society of America 20.04.2021
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Summary:Stimulated emission depletion (STED) microscopy enables the three-dimensional (3D) visualization of dynamic nanoscale structures in living cells, offering unique insights into their organization. However, 3D-STED imaging deep inside biological tissue is obstructed by optical aberrations and light scattering. We present a STED system that overcomes these challenges. Through the combination of two-photon excitation, adaptive optics, red-emitting organic dyes, and a long-working-distance water-immersion objective lens, our system achieves aberration-corrected 3D super-resolution imaging, which we demonstrate 164 µm deep in fixed mouse brain tissue and 76 µm deep in the brain of a living mouse.
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ISSN:2334-2536
2334-2536
DOI:10.1364/optica.416841