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Shy Shoham, Adi Schejter, Limor Tsur, Inna Reutsky-Gefen, Nairouz Farah; Combining holographic stimulation with cellular resolution imaging in the rodent eye. Invest. Ophthalmol. Vis. Sci. 2013;54(15):1039.
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Optical retinal prostheses for patients with outer-retinal degenerative diseases could interface directly with surviving retinal neurons in order to mimic the normal input obtained from photoreceptors in healthy retinas. Recently, we introduced an artificial photo-stimulation technique based on the projection of holographic patterns with high spatio-temporal resolution onto optogentic probes, to selectively control large retinal neuronal populations in the isolated retina. Here, we explore the ability to target single optogenetic-expressing retinal ganglion cells (RGCs) with holographic patterns at a cellular resolution, in-vivo.
For image-guided neuronal targeting, we constructed a system that combines precise spatiotemporal holographic photo-stimulation with high resolution fundus imaging. The system is also integrated with a multiphoton microscope to enable functional imaging of the responses to artificial stimulation.
The system was utilized to acquire both brightfield and fluorescence fundus images of mice and rats in-vivo, enabling the identification of single fluorescent RGCs for stimulation. Holographic patterns were projected onto the rodents’ retinas and imaged. The stimulation spot diameter is sufficient for cellular targeting using patterned photo-stimulation.
Our system enables single-cell resolved patterned holographic photo-stimulation of RGCs in-vivo which will enable the further development of a novel optical retinal prosthesis.
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