bioRxivBiophysics
3D super-resolution deep-tissue imaging in living mice
M. G. M. Velasco, M. Zhang, J. Antonello, P. Yuan, E. S. Allgeyer, D. May, O. M'Saad, P. Kidd, A. E. S. Barentine, V. Greco, J. Grutzendler, M. J. Booth, J. Bewersdorf
Abstract
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 {micro}m deep in fixed mouse brain tissue and 76 {micro}m deep in the brain of a living mouse.
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