In situ autofluorescence lifetime assay of a photoreceptor stimulus response in mouse retina and human retinal organoids
Year: 2022
Type: article
Abstract: Photoreceptors are the key functional cell types responsible for the initiation of vision in the retina. Phototransduction involves isomerization and conversion of vitamin A compounds, known as retinoids, and their recycling through the visual cycle. We demonstrate a functional readout of the visual cycle in photoreceptors within stem cell-derived retinal organoids and mouse retinal explants based on spectral and lifetime changes in autofluorescence of the visual cycle retinoids after exposure to light or chemical stimuli. We also apply a simultaneous two- and three-photon excitation method that provides specific signals and increases contrast between these retinoids, allowing for reliable detection of their presence and conversion within photoreceptors. This multiphoton imaging technique resolves the slow dynamics of visual cycle reactions and can enable high-throughput functional screening of retinal tissues and organoid cultures with single-cell resolution.
Source: Biomedical Optics Express
DOI: 10.1364/boe.455783
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: gold
Estimated APC paid: $1,913
Funders: Research to Prevent Blindness, Muskingum County Community Foundation, Carl Marshall and Mildred Almen Reeves Foundation, Retina Research Foundation, National Heart, Lung, and Blood Institute, National Eye Institute, National Cancer Institute, National Institute of Biomedical Imaging and Bioengineering, Eunice Kennedy Shriver National Institute of Child Health and Human Development
Awards: Single Cell Profiling To Define Biomarkers Of Photoreceptor Dysfunction After Gene Editing Within PSC-Derived Organoids, Mechanisms of Retinogenesis in Human Stem Cells, Waisman Intellectual and Developmental Disabilities Research Center, Optical imaging of pancreas cancer organoids for drug development and personalized treatment, Cellular level optical metabolic imaging to predict drug response in cancer, (PQ7) Quantitative in vivo optical imaging of tumor heterogeneity, Single Cell Profiling To Define Biomarkers Of Photoreceptor Dysfunction After Gene Editing Within PSC-Derived Organoids, Single Cell Profiling To Define Biomarkers Of Photoreceptor Dysfunction After Gene Editing Within PSC-Derived Organoids
OpenAlex ID: W4280501948