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Large scale interrogation of retinal cell functions by 1-photon light-sheet microscopy

View ORCID ProfileSuva Roy, Depeng Wang, View ORCID ProfileAndra M. Rudzite, Benjamin Perry, View ORCID ProfileMiranda L. Scalabrino, View ORCID ProfileMishek Thapa, View ORCID ProfileYiyang Gong, View ORCID ProfileAlexander Sher, View ORCID ProfileGreg D. Field
doi: https://doi.org/10.1101/2022.09.26.508527
Suva Roy
1Department of Neurobiology, Duke University School of Medicine, Durham, NC, USA
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  • For correspondence: suva.roy@duke.edu
Depeng Wang
2Department of Biomedical Engineering, Duke University, Durham, NC, USA
3College of Energy and Power Engineering, Nanjing University of Aeronautics and Astronautics, Nanjing, China
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Andra M. Rudzite
1Department of Neurobiology, Duke University School of Medicine, Durham, NC, USA
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Benjamin Perry
2Department of Biomedical Engineering, Duke University, Durham, NC, USA
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Miranda L. Scalabrino
1Department of Neurobiology, Duke University School of Medicine, Durham, NC, USA
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Mishek Thapa
1Department of Neurobiology, Duke University School of Medicine, Durham, NC, USA
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Yiyang Gong
2Department of Biomedical Engineering, Duke University, Durham, NC, USA
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Alexander Sher
4Santa Cruz Institute for Particle Physics, University of California, Santa Cruz, Santa Cruz, CA, USA
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Greg D. Field
1Department of Neurobiology, Duke University School of Medicine, Durham, NC, USA
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Abstract

Visual processing in the retina depends on the collective activity of large ensembles of neurons organized in different layers. Current techniques for measuring activity of layer-specific neural ensembles rely on expensive pulsed infrared lasers to drive 2-photon activation of calcium-dependent fluorescent reporters. Here, we present a 1-photon light-sheet imaging system that can measure the activity in hundreds of ex vivo retinal neurons over a large field of view while simultaneously presenting visual stimuli. This allowed for a reliable functional classification of different retinal ganglion cell types. We also demonstrate that the system has sufficient resolution to image calcium entry at individual synaptic release sites across the axon terminals of dozens of simultaneously imaged bipolar cells. The simple design, a large field of view, and fast image acquisition, make this a powerful system for high-throughput and high-resolution measurements of retinal processing at a fraction of the cost of alternative approaches.

Competing Interest Statement

The authors have declared no competing interest.

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The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. All rights reserved. No reuse allowed without permission.
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Posted September 28, 2022.
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Large scale interrogation of retinal cell functions by 1-photon light-sheet microscopy
Suva Roy, Depeng Wang, Andra M. Rudzite, Benjamin Perry, Miranda L. Scalabrino, Mishek Thapa, Yiyang Gong, Alexander Sher, Greg D. Field
bioRxiv 2022.09.26.508527; doi: https://doi.org/10.1101/2022.09.26.508527
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Large scale interrogation of retinal cell functions by 1-photon light-sheet microscopy
Suva Roy, Depeng Wang, Andra M. Rudzite, Benjamin Perry, Miranda L. Scalabrino, Mishek Thapa, Yiyang Gong, Alexander Sher, Greg D. Field
bioRxiv 2022.09.26.508527; doi: https://doi.org/10.1101/2022.09.26.508527

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