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

• Published in: bioRxiv
• Main Authors: Velasco, Mary Grace M, Zhang, Mengyang, Antonello, Jacopo, Yuan, Peng, Allgeyer, Edward S, May, Dennis, Ons M'saad, Kidd, Phylicia, Barentine, Andrew E S, Greco, Valentina, Grutzendler, Jaime, Booth, Martin J, Bewersdorf, Joerg
• Format: Paper
• Language: English
• Published: Cold Spring Harbor Cold Spring Harbor Laboratory Press 02.10.2019; Cold Spring Harbor Laboratory
• Edition: 1.1
• Subjects: Biophysics; Light scattering; Neuroimaging; Optics
• ISSN: 2692-8205; 2692-8205
• DOI: 10.1101/790212

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 2-photon excitation, adaptive optics, far-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.