Abstract
Oblique scanning laser ophthalmoscopy (oSLO) is a recently developed technique to provide three-dimensional volumetric fluorescence imaging in retinas over a large field of view, without the need for depth sectioning. In this study, we present volumetric fluorescein angiography (vFA) at 200 B-scans per second in mouse retina in vivo by oSLO. By using a low-cost industrial CMOS camera, imaging speed was improved to 2 volumes per second, ∼10 times more than our previous results. Enabled by the volumetric imaging, we visualized hemodynamics at single capillary level in a depth-dependent manner, and provided methods to quantify capillary hematocrit, absolute capillary blood flow speed, and detection of capillary flow stagnancy and stalling at different vascular layers. The quantitative metrics for capillary hemodynamics enhanced by volumetric imaging can offer valuable insight into vision science and retinal pathologies.
Highlights
Fluorescein angiography (FA) is a major imaging method in ophthalmology, for care of retinal vascular diseases such as diabetic retinopathy (DR) and age-related macular degeneration (AMD)
For the first time to our knowledge, we demonstrated in vivo volumetric fluorescein angiography (vFA) imaging on mouse retina by Oblique scanning laser ophthalmoscopy (oSLO) with significantly increased the imaging speed, achieving vFA at 2 volumes per second
In order to analyze vFA images at different vascular layers, we referenced the depth to outer plexiform layer (OPL) where the deep capillary plexus is located
Summary
Fluorescein angiography (FA) is a major imaging method in ophthalmology, for care of retinal vascular diseases such as diabetic retinopathy (DR) and age-related macular degeneration (AMD). An oblique imaging system is aligned such that a camera sensor is conjugated with the oblique light sheet to capture a cross-sectional FA image [8,9,10,11,12]. This mechanism uniquely enabled vFA by only one raster scan, without the need for depth sectioning. The improved high-speed oSLO allows us to capture and quantify hemodynamics at individual capillary level in 3D
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