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