Cellular Resolution Panretinal Imaging of Optogenetic Probes Using a Simple Funduscope
| Autor: | Nairouz Farah, Yishay Falick, Inna Reutsky-Gefen, Shy Shoham, Limor Tsur, Adi Schejter |
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| Rok vydání: | 2012 |
| Předmět: |
Physics
0303 health sciences Pathology medicine.medical_specialty Retina genetic structures Biomedical Engineering Channelrhodopsin Optogenetics Fundus (eye) Retinal ganglion Article eye diseases Funduscope 03 medical and health sciences Ophthalmology 0302 clinical medicine medicine.anatomical_structure In vivo medicine Retinal imaging sense organs 030217 neurology & neurosurgery 030304 developmental biology Biomedical engineering |
| Zdroj: | Translational Vision Science & Technology |
| ISSN: | 2164-2591 |
| Popis: | PURPOSE To acquire and characterize cellular-resolved in vivo fluorescence images of optogenetic probes expressed in rodent retinal ganglion cells, by adapting a low-cost and simple fundus system based on a topical endoscope. METHODS A custom endoscope-based fundus system was constructed (adapted from the design of Paques and colleagues). Bright field and fluorescence images were acquired from head-fixed transgenic mice expressing Channelrhodopsin2-eYFP, and Sprague Dawley rats virally transfected with the optogenetic probe GCaMP3. Images were compared to in vitro images of the same structures and were analyzed. RESULTS The fundus system provides high-quality, high-resolution fluorescence images of the eye fundus that span the whole retina. The images allow resolving individual cells and axon bundles in the Channelrhodopsin2-eYFP mice and cellular-scale structures in the GCaMP3 expressing rats. The resolution in mouse eyes was estimated to be better than 20 μm (full width at half maximum) and is only marginally dependent on movement-related blurring. CONCLUSIONS The fluorescence-endoscopy fundus system provides a powerful yet simple and widely accessible tool for obtaining cellular resolved fluorescent images of optogenetic and other fluorescent probes. TRANSLATIONAL RELEVANCE The new system could prove to be a basic tool for non-invasive in vivo small animal retinal imaging in a wide array of translational vision applications, including the tracking of fluorescently tagged cells and the expression of gene-therapy and optogenetic vectors. |
| Databáze: | OpenAIRE |
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