Abstract
Two-photon excited fluorescence (TPEF) imaging of the retina is a developing technique that provides non-invasive compound-specific measurements from the retina. In this report, we demonstrate high-resolution TPEF imaging of the mouse retina using sensorless adaptive optics (SAO) and optical coherence tomography (OCT). A single near-infrared light source was used for simultaneous multi-modal imaging with OCT and TPEF. The image-based SAO could be performed using the en face OCT or the TPEF for aberration correction. Our results demonstrate OCT and TPEF for angiography. Also, we demonstrate non-invasive cellular-resolution imaging of fluorescently labelled cells and the Retinal Pigment Epithelium (RPE) mosaic.
Highlights
Non-invasive retinal imaging is a valuable tool that is used in both clinical and preclinical vision research to aid the development of novel therapies for preventing irreversible vision loss
The murine vasculature of the inner retina is stacked in three distinct layers, including the Outer Plexiform Layer (OPL), Inner Plexiform Layer (IPL), and Nerve Fiber Layer (NFL)
We demonstrate the ability to image GFP labelled retinal ganglion cells (RGC) of a transgenic mouse strain (Tg(Thy1-EGFP)MJrs/J) with sensorless adaptive optics (SAO)-Two-Photon Excited Fluorescence (TPEF) and we compared the images to single photon excited fluorescence (SPEF) with SAO
Summary
Non-invasive retinal imaging is a valuable tool that is used in both clinical and preclinical vision research to aid the development of novel therapies for preventing irreversible vision loss. More sensitive assessment of the physiological and biochemical processes within the retina could be used to detect earlier signs of disease in order to preserve sight [1,2]. Fluorescence can be used to image many biomarkers, since fluorophores can be added to the retina to provide contrast or fluorophores that are intrinsic to the retina can be used for measurements. For many fluorophores intrinsic to the retina, the single-photon excitation is in the ultraviolet (UV) range and the fluorescence cannot be excited through the eye of many species due to the ocular transmission window [6,7]. Imaging the retina non-invasively with Two-Photon Excited Fluorescence (TPEF) could enable novel in vivo studies of disease and retinal physiology [6–9]. Imaging the retina with near-infrared (NIR) light has advantages since the retina is less sensitive to NIR than visible light. NIR light is less scattered within biological tissue than the equivalent visible light required to excite the same fluorophores [10,11]
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