Two-Photon Autofluorescence Imaging Reveals Cellular Structures Throughout the Retina of the Living Primate Eye

Investigative Ophthalmology & Visual Science
Robin SharmaJennifer J Hunter

Abstract

Although extrinsic fluorophores can be introduced to label specific cell types in the retina, endogenous fluorophores, such as NAD(P)H, FAD, collagen, and others, are present in all retinal layers. These molecules are a potential source of optical contrast and can enable noninvasive visualization of all cellular layers. We used a two-photon fluorescence adaptive optics scanning light ophthalmoscope (TPF-AOSLO) to explore the native autofluorescence of various cell classes spanning several layers in the unlabeled retina of a living primate eye. Three macaques were imaged on separate occasions using a custom TPF-AOSLO. Two-photon fluorescence was evoked by pulsed light at 730 and 920 nm excitation wavelengths, while fluorescence emission was collected in the visible range from several retinal layers and different locations. Backscattered light was recorded simultaneously in confocal modality and images were postprocessed to remove eye motion. All retinal layers yielded two-photon signals and the heterogeneous distribution of fluorophores provided optical contrast. Several structural features were observed, such as autofluorescence from vessel walls, Müller cell processes in the nerve fibers, mosaics of cells in the ganglion cell ...Continue Reading

Citations

Feb 19, 2017·Vision Research·Susana MarcosLawrence C Sincich
Jan 24, 2017·Progress in Retinal and Eye Research·Melissa K JonesShaomei Wang
Dec 18, 2018·Investigative Ophthalmology & Visual Science·Christina SchwarzJennifer J Hunter
Dec 21, 2018·Investigative Ophthalmology & Visual Science·Jia QinJacque L Duncan
Sep 17, 2019·Annual Review of Vision Science·Jennifer J HunterJesse B Schallek
Feb 19, 2019·Biomedical Optics Express·Sarah WaltersWilliam H Merigan
Aug 29, 2019·Biomedical Engineering Letters·Harsh RanawatNirmal Mazumder
Jul 23, 2016·Biomedical Optics Express·Nathan S AlexanderKrzysztof Palczewski
Jan 5, 2017·Proceedings of the National Academy of Sciences of the United States of America·Ethan A RossiDavid R Williams
Jun 20, 2018·Cells·Timothy E YapMaria Francesca Cordeiro
Jul 20, 2019·Alzheimer's Research & Therapy·Jurre den HaanKeir X X Yong
Aug 31, 2019·Biomedical Optics Express·Gopal Swamy JayabalanJoseph T Fan
Dec 27, 2016·Biomedical Optics Express·Christina SchwarzJennifer J Hunter
Dec 2, 2020·Eye·Engin AkyolAndrew J Lotery
Mar 25, 2021·Investigative Ophthalmology & Visual Science·Zhuolin LiuDaniel X Hammer
Jun 11, 2021·Translational Vision Science & Technology·Kira H LinAla Moshiri
Dec 1, 2021·The Journal of Clinical Investigation·Jakub BoguslawskiMaciej Wojtkowski

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Methods Mentioned

BETA
conventional imaging
Fluorescence

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