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

View ORCID ProfileMary Grace M. Velasco, Mengyang Zhang, View ORCID ProfileJacopo Antonello, Peng Yuan, View ORCID ProfileEdward S. Allgeyer, Dennis May, Ons M’Saad, Phylicia Kidd, View ORCID ProfileAndrew E. S. Barentine, Valentina Greco, Jaime Grutzendler, Martin J. Booth, View ORCID ProfileJoerg Bewersdorf
doi: https://doi.org/10.1101/790212
Mary Grace M. Velasco
1Department of Biomedical Engineering, Yale School of Engineering & Applied Science, CT 06520, USA
2Department of Cell Biology, Yale School of Medicine, CT 06520, USA
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  • ORCID record for Mary Grace M. Velasco
Mengyang Zhang
3Interdepartmental Neuroscience Program, Yale School of Medicine, CT 06520, USA
4Department of Neuroscience, Yale School of Medicine, CT 06520, USA
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Jacopo Antonello
5Department of Engineering Science, University of Oxford, Oxford OX1 3PJ, UK
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Peng Yuan
4Department of Neuroscience, Yale School of Medicine, CT 06520, USA
6Department of Neurology, Yale School of Medicine, CT 06520, USA
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Edward S. Allgeyer
2Department of Cell Biology, Yale School of Medicine, CT 06520, USA
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Dennis May
7Department of Genetics, Yale School of Medicine, CT 06520, USA
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Ons M’Saad
1Department of Biomedical Engineering, Yale School of Engineering & Applied Science, CT 06520, USA
2Department of Cell Biology, Yale School of Medicine, CT 06520, USA
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Phylicia Kidd
2Department of Cell Biology, Yale School of Medicine, CT 06520, USA
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Andrew E. S. Barentine
1Department of Biomedical Engineering, Yale School of Engineering & Applied Science, CT 06520, USA
2Department of Cell Biology, Yale School of Medicine, CT 06520, USA
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Valentina Greco
2Department of Cell Biology, Yale School of Medicine, CT 06520, USA
7Department of Genetics, Yale School of Medicine, CT 06520, USA
8Department of Dermatology, Yale Stem Cell Center, Yale Cancer Center, Yale School of Medicine, CT 06520, USA
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Jaime Grutzendler
3Interdepartmental Neuroscience Program, Yale School of Medicine, CT 06520, USA
4Department of Neuroscience, Yale School of Medicine, CT 06520, USA
6Department of Neurology, Yale School of Medicine, CT 06520, USA
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Martin J. Booth
5Department of Engineering Science, University of Oxford, Oxford OX1 3PJ, UK
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Joerg Bewersdorf
1Department of Biomedical Engineering, Yale School of Engineering & Applied Science, CT 06520, USA
2Department of Cell Biology, Yale School of Medicine, CT 06520, USA
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  • ORCID record for Joerg Bewersdorf
  • For correspondence: joerg.bewersdorf@yale.edu
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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 µm deep in fixed mouse brain tissue and 76 µm deep in the brain of a living mouse.

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Posted October 02, 2019.
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3D super-resolution deep-tissue imaging in living mice
Mary Grace M. Velasco, Mengyang Zhang, Jacopo Antonello, Peng Yuan, Edward S. Allgeyer, Dennis May, Ons M’Saad, Phylicia Kidd, Andrew E. S. Barentine, Valentina Greco, Jaime Grutzendler, Martin J. Booth, Joerg Bewersdorf
bioRxiv 790212; doi: https://doi.org/10.1101/790212
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3D super-resolution deep-tissue imaging in living mice
Mary Grace M. Velasco, Mengyang Zhang, Jacopo Antonello, Peng Yuan, Edward S. Allgeyer, Dennis May, Ons M’Saad, Phylicia Kidd, Andrew E. S. Barentine, Valentina Greco, Jaime Grutzendler, Martin J. Booth, Joerg Bewersdorf
bioRxiv 790212; doi: https://doi.org/10.1101/790212

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