The AOSLO can resolve the smallest foveal cones in certain eyes. Although myopia causes retinal stretching in the fovea, its effect within the foveola is confounded by factors other than cone density that have high levels of intersubject variability.
It is often difficult to find consistent changes in the retinal microvasculature due to large intersubject variability. However, with a novel application of AOSLO imaging, it is possible to visualize parafoveal capillaries and identify AV channels noninvasively. AV channels are disrupted in type 2 diabetes, even before the onset of diabetic retinopathy.
We demonstrate the capability of a new generation adaptive optics scanning laser ophthalmoscope (AOSLO) to resolve cones and rods in normal subjects, and confirm our findings by comparing cone and rod spacing with published histology measurements. Cone and rod spacing measurements are also performed on AOSLO images from two different diseased eyes, one affected by achromatopsia and the other by acute zonal occult outer retinopathy (AZOOR). The potential of AOSLO technology in the study of these and other retinal diseases is illustrated.
We demonstrate projection of highly stabilized, aberration-corrected stimuli directly onto the retina by means of real-time retinal image motion signals in combination with high speed modulation of a scanning laser. In three subjects with good fixation stability, stimulus location accuracy averaged 0.26 arcminutes or approximately 1.3 microns, which is smaller than the cone-to-cone spacing at the fovea. We also demonstrate real-time correction for image distortions in adaptive optics scanning laser ophthalmoscope (AOSLO) with an intraframe accuracy of about 7 arcseconds.
We demonstrate a high-speed, image-based tracking scanning laser ophthalmoscope (TSLO) that can
provide high fidelity structural images, real-time eye tracking and targeted stimulus delivery. The
system was designed for diffraction-limited performance over an 8° field of view (FOV) and
operates with a flexible field of view of 1°–5.5°. Stabilized videos of the
retina were generated showing an amplitude of motion after stabilization of 0.2 arcmin or less
across all frequencies. In addition, the imaging laser can be modulated to place a stimulus on a
targeted retinal location. We show a stimulus placement accuracy with a standard deviation less than
1 arcmin. With a smaller field size of 2°, individual cone photoreceptors were clearly
visible at eccentricities outside of the fovea.
Adaptive Optics Scanning Laser Ophthalmoscopy was used to noninvasively acquire videos of single-file flow through live human retinal parafoveal capillaries. Videos were analyzed offline to investigate capillary flow dynamics. Certain capillaries accounted for a clear majority of leukocyte traffic (Leukocyte-Preferred-Paths, LPPs), while other capillaries primarily featured plasma gap flow (Plasma-Gap-Capillaries, PGCs). LPPs may serve as a protective mechanism to prevent inactivated leukocytes from entering exchange capillaries, and PGCs may serve as relief valves to minimize flow disruption due to the presence of a leukocyte in a neighboring LPP.
A special challenge arises when pursuing multi-wavelength imaging of retinal tissue in vivo, because the eye’s optics must be used as the main focusing elements, and they introduce significant chromatic dispersion. Here we present an image-based method to measure and correct for the eye’s transverse chromatic aberrations rapidly, non-invasively, and with high precision. We validate the technique against hyperacute psychophysical performance and the standard chromatic human eye model. In vivo correction of chromatic dispersion will enable confocal multi-wavelength images of the living retina to be aligned, and allow targeted chromatic stimulation of the photoreceptor mosaic to be performed accurately with sub-cellular resolution.
Purpose
To establish adaptive optics scanning laser ophthalmoscopy (AOSLO) as a method to detect and characterize microscopic signs of diabetic retinopathy, in capillaries and cone photoreceptors in the parafovea.
Methods
Recently, AOSLO has enabled noninvasive assessment of photoreceptors, capillaries, and leukocytes in the retinas of live human subjects. Repeated application of AOSLO imaging along with comparison to fluorescein angiography was used to track individual capillaries near the foveal avascular zone (FAZ) from one eye affected with severe non-proliferative diabetic retinopathy. Fluorescein angiography was used to identify clinical signs of diabetic retinopathy, such as microaneurysms and intra-retinal microvascular abnormalities (IRMAs), and corresponding regions were imaged and assessed using the AOSLO. In addition, the structural integrity of photoreceptors and the spatial distribution of leukocytes around the parafoveal capillary network were quantitatively assessed.
Results
Capillaries and cone photoreceptors were visualized using the AOSLO, without the use of injected contrast agents. Although the majority of capillaries were stable over a period of 16 months, one capillary at the edge of the FAZ dropped out, leading to a small, but significant increase in FAZ size. Longitudinal assessment of the capillaries also showed microaneurysm formation and disappearance, as well as the formation of tiny capillary bends similar in appearance to IRMAs. The leukocytes in the capillary network were found to preferentially travel through the same routes in all four visits, suggesting that these channels are robust against small changes to the surrounding capillaries. In this eye, cone photoreceptor spacing was increased in the fovea when compared to normal data, but stable across all visits.
Conclusions
AOSLO imaging can be used to longitudinally track capillaries, leukocytes and photoreceptors in diabetic retinopathy. Capillary changes that can be detected include dropout of individual capillaries, as well as formation and disappearance of microaneurysms.
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