Pavan Tiruveedhula

Observation of cone and rod photoreceptors in normal subjects and patients using a new generation adaptive optics scanning laser ophthalmoscope

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.

Intersubject Variability of Foveal Cone Photoreceptor Density in Relation to Eye Length

PURPOSE Adaptive optics scanning laser ophthalmoscopy (AOSLO) under optimized wavefront correction allows for routine imaging of foveal cone photoreceptors. The intersubject variability of foveal cone density was measured and its relation to eye length evaluated. METHODS AOSLO was used to image 18 healthy eyes with axial lengths from 22.86 to 28.31 mm. Ocular biometry and an eye model were used to estimate the retinal magnification factor. Individual cones in the AOSLO images were labeled, and the locations were used to generate topographic maps representing the spatial distribution of density. Representative retinal (cones/mm(2)) and angular (cones/deg(2)) cone densities at specific eccentricities were calculated from these maps. RESULTS The entire foveal cone mosaic was resolved in four eyes, whereas the cones within 0.03 mm eccentricity remained unresolved in most eyes. The preferred retinal locus deviated significantly (P < 0.001) from the point of peak cone density for all except one individual. A significant decrease in retinal density (P < 0.05) with increasing axial length was observed at 0.30 mm eccentricity but not closer. Longer, more myopic eyes generally had higher angular density near the foveal center than the shorter eyes, but by 1°, this difference was nullified by retinal expansion, and so angular densities across all eyes were similar. CONCLUSIONS 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.

Retinally stabilized cone-targeted stimulus delivery

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.

Disruption of the Retinal Parafoveal Capillary Network in Type 2 Diabetes before the Onset of Diabetic Retinopathy

PURPOSE To establish, using adaptive optics scanning laser ophthalmoscopy (AOSLO), that the retinal parafoveal capillary network is altered before the onset of diabetic retinopathy in adult patients with type 2 diabetes. METHODS AOSLO videos were acquired in the parafoveal region of one eye from control subjects and from patients with type 2 diabetes and no retinopathy. Detailed images of the parafoveal capillary network were generated with custom motion contrast enhancement algorithms. The combination of AOSLO images and videos enabled the simultaneous assessment of several features of the parafoveal capillary network. Arteriovenous (AV) channels were identified by finding the least tortuous capillary channels connecting terminal arterioles to postcapillary venules. Measures of capillary dropout and capillary hemodynamics were also quantified. RESULTS The average tortuosity of AV channels was 26% higher in patients with type 2 diabetes when compared with controls, even though there were no signs of diabetic retinopathy in any of the eyes that were assessed (P < 0.05). In addition, the metrics of capillary dropout showed small changes (between 3% and 7%), leukocyte speed 14% lower, and pulsatility 25% higher, but none of these differences was statistically significant. CONCLUSIONS 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.

Resolving single cone inputs to visual receptive fields.

With the current techniques available for mapping receptive fields, it is impossible to resolve the contribution of single cone photoreceptors to the response of central visual neurons. Using adaptive optics to correct for ocular aberrations, we delivered micron-scale spots of light to the receptive field centers of neurons in the macaque lateral geniculate nucleus. Parvocellular LGN neurons mapped in this manner responded with high reliability to stimulation of single cones.

Characterization of single-file flow through human retinal parafoveal capillaries using an adaptive optics scanning laser ophthalmoscope.

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.

Real-time eye motion correction in phase-resolved OCT angiography with tracking SLO

In phase-resolved OCT angiography blood flow is detected from phase changes in between A-scans that are obtained from the same location. In ophthalmology, this technique is vulnerable to eye motion. We address this problem by combining inter-B-scan phase-resolved OCT angiography with real-time eye tracking. A tracking scanning laser ophthalmoscope (TSLO) at 840 nm provided eye tracking functionality and was combined with a phase-stabilized optical frequency domain imaging (OFDI) system at 1040 nm. Real-time eye tracking corrected eye drift and prevented discontinuity artifacts from (micro)saccadic eye motion in OCT angiograms. This improved the OCT spot stability on the retina and consequently reduced the phase-noise, thereby enabling the detection of slower blood flows by extending the inter-B-scan time interval. In addition, eye tracking enabled the easy compounding of multiple data sets from the fovea of a healthy volunteer to create high-quality eye motion artifact-free angiograms. High-quality images are presented of two distinct layers of vasculature in the retina and the dense vasculature of the choroid. Additionally we present, for the first time, a phase-resolved OCT angiogram of the mesh-like network of the choriocapillaris containing typical pore openings.

Subclinical capillary changes in non-proliferative diabetic retinopathy.

Purpose. To establish adaptive optics scanning laser ophthalmoscopy as a method to detect and characterize microscopic signs of diabetic retinopathy in capillaries and cone photoreceptors in the parafovea. Methods. Recently, adaptive optics scanning laser ophthalmoscope (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 intraretinal microvascular abnormalities, 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 intraretinal microvascular abnormalities. 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 with 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.

High-speed, image-based eye tracking with a scanning laser ophthalmoscope.

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.

Real-time eye motion compensation for OCT imaging with tracking SLO

Fixational eye movements remain a major cause of artifacts in optical coherence tomography (OCT) images despite the increases in acquisition speeds. One approach to eliminate the eye motion is to stabilize the ophthalmic imaging system in real-time. This paper describes and quantifies the performance of a tracking OCT system, which combines a phase-stabilized optical frequency domain imaging (OFDI) system and an eye tracking scanning laser ophthalmoscope (TSLO). We show that active eye tracking minimizes artifacts caused by eye drift and micro saccades. The remaining tracking lock failures caused by blinks and large saccades generate a trigger signal which signals the OCT system to rescan corrupted B-scans. Residual motion artifacts in the OCT B-scans are reduced to 0.32 minutes of arc (~1.6 µm) in an in vivo human eye enabling acquisition of high quality images from the optic nerve head and lamina cribrosa pore structure.