An in situ autofluorescent assay of photoreceptor stimulus response in mouse retina and human retinal organoids

Kayvan Samimi; Bikash Pattnaik; Elizabeth E. Capowski; Katherine P. Mueller; Amr A. Abdeen; Krishanu Saha; David M. Gamm; Melissa C. Skala

doi:10.1117/12.2609771

7 March 2022 An in situ autofluorescent assay of photoreceptor stimulus response in mouse retina and human retinal organoids

Kayvan Samimi, Bikash Pattnaik, Elizabeth E. Capowski, Katherine P. Mueller, Amr A. Abdeen, Krishanu Saha, David M. Gamm, Melissa C. Skala

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Proceedings Volume PC11965, Multiphoton Microscopy in the Biomedical Sciences XXII; PC119650G (2022) https://doi.org/10.1117/12.2609771
Event: SPIE BiOS, 2022, San Francisco, California, United States

Abstract

Human pluripotent stem cell (hPSC)-derived retinal cell culture holds great promise for treatment of retinal diseases that cause blindness. However, high-throughput non-invasive functional assays are needed for rapid optimization and quality control of these stem cell culture systems. Here we use simultaneous two- and three-photon excited fluorescence lifetime imaging microscopy (FLIM) to characterize the autofluorescent signature of light response in photoreceptor cells that originates from the fluorescent vitamin A compounds (i.e., retinoids) of the visual cycle. This multiphoton microscopy technique resolves the dynamics of visual pigment photobleaching and recycling following light exposure, including conversion of different retinoids to one another.

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Kayvan Samimi, Bikash Pattnaik, Elizabeth E. Capowski, Katherine P. Mueller, Amr A. Abdeen, Krishanu Saha, David M. Gamm, and Melissa C. Skala "An in situ autofluorescent assay of photoreceptor stimulus response in mouse retina and human retinal organoids", Proc. SPIE PC11965, Multiphoton Microscopy in the Biomedical Sciences XXII, PC119650G (7 March 2022); https://doi.org/10.1117/12.2609771

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KEYWORDS

Retina

Luminescence

Absorption

Fluorescence lifetime imaging

Multiphoton microscopy

Pulsed laser operation

Stem cells

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