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

利用单光子光片显微镜对视网膜细胞功能进行大规模研究

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作者:Suva Roy ,Depeng Wang ,Andra M Rudzite ,Benjamin Perry ,Miranda L Scalabrino ,Mishek Thapa ,Yiyang Gong ,Alexander Sher ,Greg D Field

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. We present a 1-photon light-sheet imaging system that can measure the activity in hundreds of neurons in the ex vivo retina over a large field of view while presenting visual stimuli. This allows for a reliable functional classification of different retinal 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, 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. Keywords: GCaMP; bipolar cells; calcium imaging; cell type; functional classification; immunohistochemistry; light-sheet imaging; retina; retinal ganglion cells; synapse.

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