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

Velasco, M. G. M.; Zhang, M.; Antonello, J.; Yuan, P.; Allgeyer, E. S.; May, D.; M'Saad, O.; Kidd, P.; Barentine, A. E. S.; Greco, V.; Grutzendler, J.; Booth, M. J.; Bewersdorf, J.

doi:10.1101/790212

bioRxiv · 10.1101/790212

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

Velasco, M. G. M.·Zhang, M.·Antonello, J.·Yuan, P.·Allgeyer, E. S.·May, D.·M'Saad, O.·Kidd, P.·Barentine, A. E. S.·Greco, V.·Grutzendler, J.·Booth, M. J.·Bewersdorf, J.

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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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Velasco, M. G. M., Zhang, M., Antonello, J., Yuan, P., Allgeyer, E. S., May, D., M'Saad, O., Kidd, P., Barentine, A. E. S., Greco, V., Grutzendler, J., Booth, M. J., Bewersdorf, J.. 2019-10-02. 3D super-resolution deep-tissue imaging in living mice. https://doi.org/10.1101/790212

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