Automatic montaging of adaptive optics SLO retinal images based on graph theory.

We present a fully automatic montage pipeline for adaptive optics SLO retinal images. It contains a flexible module to estimate the translation between pairwise images. The user can change modules to accommodate the alignment of the dataset using the most appropriate alignment technique, provided that it estimates the translation between image pairs and provides a quantitative confidence metric for the match between 0 and 1. We use these pairwise comparisons and associated metrics to construct a graph where nodes represent frames and edges represent the overlap relations. We use a small diameter spanning tree to determine the best pairwise alignment for each image based on the entire set of image relations. The final stage of the pipeline is a blending module that uses dynamic programming to improve the smoothness of the transition between frames. Data sets ranging from 26 to 119 images were obtained from individuals aged 24 to 81 years with a mix of visually normal control eyes and eyes with glaucoma or diabetes. The resulting automatically generated montages were qualitatively and quantitatively compared to results from semi-automated alignment. Data sets were specifically chosen to include both high quality and medium quality data. The results obtained from the automatic method are comparable or better than results obtained by an experienced operator performing semi-automated montaging. For the plug-in pairwise alignment module, we tested a technique that utilizes SIFT + RANSAC, Normalized cross-correlation (NCC) and a combination of the two. This pipeline produces consistent results not only on outer retinal layers, but also on inner retinal layers such as a nerve fiber layer or images of the vascular complexes, even when images are not of excellent quality.

Retinal Arteriolar Wall Remodeling in Diabetes Captured With AOSLO.

Purpose: Adaptive optics scanning laser ophthalmoscopy (AOSLO) enables the visualization and measurement of the retinal microvasculature structure in humans. We investigated the hypothesis that diabetes mellitus (DM) induces remodeling to the wall structure in small retinal arterioles. These alterations may allow better understanding of vascular remodeling in DM. Methods: We imaged retinal arterioles in one eye of 48 participants (26 with DM and 22 healthy controls) with an AOSLO. Structural metrics of 274 arteriole segments (203 with DM and 71 healthy controls) ≤ 50 µm in outer diameter (OD) were quantified and we compared differences in wall thickness (WT), wall-to-lumen ratio (WLR), inner diameter (ID), OD, and arteriolar index ratio (AIR) between controls and participants with DM. We also compared the individual AIR (iAIR) in groups of individuals. Results: The WLR, WT, and AIRs were significantly different in the arteriole segments of DM participants (P < 0.001). The iAIR was significantly deviated in the DM group (P < 0.001) and further division of the participants with DM into groups revealed that there was an effect of the presence of diabetic retinopathy (DR) on the iAIR (P < 0.001). Conclusions: DM induces remodeling of wall structure in small retinal arterioles and in groups of individuals. The use of AIR allows us to assess remodeling independently of vessel size in the retina and to compute an index for each individual subject. Translational Relevance: High-resolution retinal imaging allows noninvasive assessment of small retinal vessel remodeling in DM that can improve our understanding of DM and DR in living humans.

Measuring Temporal and Spatial Variability of Red Blood Cell Velocity in Human Retinal Vessels.

Purpose: The retinal circulation regulates blood flow through various internal and external factors; however, it is unclear how locally these factors act within the retinal microcirculation. We measured the temporal and spatial variability of blood velocity in small retinal vessels using a dual-beam adaptive optics scanning laser ophthalmoscope. Methods: In young healthy subjects (n = 3), temporal blood velocity variability was measured in a local vascular region consisting of an arteriole, capillary, and venule repeatedly over 2 days. Data consisted of 10 imaging periods separated into two sessions: (1) five 6-minute image acquisition periods with 30-minute breaks, and (2) five 6-minute image acquisition periods with 10-minute breaks. In another group of young healthy subjects (n = 5), spatial distribution of velocity variability was measured by imaging three capillary segments during three 2-minute conditions: (1) baseline imaging condition (no flicker), (2) full-field flicker, and (3) no flicker condition again. Results: Blood velocities were measurable in all subjects with a reliability of about 2%. The coefficient of variation (CV) was used as an estimate of the physiological variability of each vessel. Over 2 days, the average CV in arterioles was 7% (±2%); in capillaries, it was 19% (±6%); and, in venules, it was 8% (±2%). During flicker stimulation, the average capillary CV was 16% during baseline, 15% during flicker stimulation, and 18% after flicker stimulation. Conclusions: Capillaries in the human retina exhibit spatial and temporal variations in blood velocity. This inherent variation in blood velocity places limits on studying the vascular regulation of individual capillaries, and the study presented here serves as a foundation for future endeavors.

Template free eye motion correction for scanning systems.

Scanning imaging systems are susceptible to image warping in the presence of target motion occurring within the time required to acquire an individual image frame. In this Letter, we introduce the use of a dual raster scanning approach to correct for motion distortion without the need for prior knowledge of the undistorted image. In the dual scanning approach, the target is imaged simultaneously with two imaging beams from the same imaging system. The two imaging beams share a common pupil but have a spatial shift between the beams on the imaging plane. The spatial shift can be used to measure high speed events, because it measures an identical region at two different times within the time required for acquisition of a single frame. In addition, it provides accurate spatial information, since two different regions on the target are imaged simultaneously, providing an undistorted estimate of the spatial relation between regions. These spatial and temporal relations accurately measure target motion. Data from adaptive optics scanning laser ophthalmoscope (AOSLO) imaging of the human retina are used to demonstrate this technique. We apply the technique to correct the shearing of retinal images produced by eye motion. Three control subjects were measured while imaging different retinal layers and retinal locations to qualify the effectiveness of the algorithm. Since the time shift between channels is readily adjustable, this method can be tuned to match different imaging situations. The major requirement is the need to separate the two images; in our case, we used different near infrared spectral regions and dichroic filters.

Full-field flicker evoked changes in parafoveal retinal blood flow.

When retinal activity is increased by exposure to dynamic visual stimuli, blood vessels dilate and the flow of blood within vessels increases to meet the oxygen and glucose demands of the neurons. This relationship is termed 'neurovascular coupling' and it is critical for regulating control of the human retinal vasculature. In this study, we used a recently developed technique based on a dual-beam adaptive optics scanning laser ophthalmoscope to measure changes in red blood cell velocities, vessel diameter, and flow in interconnected small parafoveal retinal vessels (< 50 µm) of nine healthy participants. A full-field flicker stimulus was presented onto the retina to induce a vascular response to neural activity. Flicker stimulation increased blood velocity, vessel diameter, and therefore flow in arterioles, capillaries, and venules in all nine subjects. ANOVA and post hoc t-test showed significant increases in velocity and flow in arterioles and venules. These measurements indicate that the mechanism of neurovascular coupling systematically affects the vascular response in small retinal vessels in order to maintain hemodynamic regulation in the retina when exposed to visual stimulation, in our case flicker. Our findings may provide insight into future investigations on the impairments of neurovascular coupling from vascular diseases such as diabetic mellitus.

High-Resolution, Adaptive Optics Imaging of the Human Trabecular Meshwork In Vivo.

PURPOSE: To image the human trabecular meshwork (TM) in vivo using adaptive optics gonioscopy (AOG) with approximately 2-µm lateral resolution. METHODS: An existing Indiana University adaptive optics scanning laser ophthalmoscope was altered by adding a 12-mm button lens to a clinical gonioscopic lens allowing high-resolution imaging of the human iridocorneal angle. First an anatomic model eye was used to refine the imaging technique and then nine participants (7 controls and 2 participants with pigment dispersion syndrome) were imaged. RESULTS: All nine participants were successfully imaged without adverse events. High-resolution imaging of the human TM was achieved allowing for visualization of the TM beams, and presumed endothelial cells. Uveal meshwork beams in controls averaged 25.5 µm (range, 15.2-44.7) in diameter with pores averaging 42.6 µm (range, 22.3-51.4) while the corneoscleral meshwork pores averaged 8.9 µm (range, 7.7-12.1). Differences in appearance of the uveal and corneoscleral meshwork were noted between the two participants with pigment dispersion syndrome and the controls. These included nearly absent spacing between the beams and enlarged endothelial cells with hyperreflective areas. CONCLUSIONS: AOG allows for near cellular level resolution of the human TM in vivo. This may allow for further understanding of age-related changes that occur as well as provide a deeper understanding of medical and surgical alterations for the treatment of glaucoma. TRANSLATIONAL RELEVANCE: Further development of this approach may allow for direct measurements at a micometer level in vivo of changes that occur in the human trabecular meshwork with glaucoma and therapeutic interventions.

Distances From Capillaries to Arterioles or Venules Measured Using OCTA and AOSLO.

Purpose: To investigate distances from retinal capillaries to arterioles or venules noninvasively. Methods: An adaptive optics scanning laser ophthalmoscope (AOSLO) and optical coherence tomography angiography (OCTA) imager acquired detailed maps of retinal vasculature. Using OCTA, we quantified the distance from the edge of an arteriole or venule to the middle of the nearest capillaries (periarteriole or perivenule capillary-free zones, respectively) within the superficial vascular plexus of 20 young healthy subjects with normal axial lengths. These distances were compared to AOSLO images for three subjects. We tested the relation between the peripheral capillary-free zones and FAZ horizontal, vertical, effective diameters, and asymmetry indices in the deep vascular plexus. We examined enlargement with OCTA of capillary-free zones in a type 2 diabetic patient. Results: The periarteriole capillary-free zone (67.2 ± 25.3 µm) was readily visible and larger than the perivenule capillary-free zone (42.7 ± 14.4 µm), F(1, 998) = 771, P < 0.0001. The distance from foveal center (P = 0.003) and diameter (P = 0.048) were predictive of perivenule capillary-free zone values. OCTA and AOSLO corresponded for arterioles. FAZ effective diameter was positively associated with asymmetry indices, r = 0.49, P = 0.028, but not peripheral capillary-free zones, although focal enlargements were found in a diabetic patient. Conclusions: For normal retinas, periarteriole and perivenule capillary-free zones are readily visible with OCTA and AOSLO. Periarteriole capillary-free zones were larger, consistent with arterioles carrying oxygen rich blood that diffuses to support the retina.

Adaptive optics imaging of the human retina.

Adaptive Optics (AO) retinal imaging has provided revolutionary tools to scientists and clinicians for studying retinal structure and function in the living eye. From animal models to clinical patients, AO imaging is changing the way scientists are approaching the study of the retina. By providing cellular and subcellular details without the need for histology, it is now possible to perform large scale studies as well as to understand how an individual retina changes over time. Because AO retinal imaging is non-invasive and when performed with near-IR wavelengths both safe and easily tolerated by patients, it holds promise for being incorporated into clinical trials providing cell specific approaches to monitoring diseases and therapeutic interventions. AO is being used to enhance the ability of OCT, fluorescence imaging, and reflectance imaging. By incorporating imaging that is sensitive to differences in the scattering properties of retinal tissue, it is especially sensitive to disease, which can drastically impact retinal tissue properties. This review examines human AO retinal imaging with a concentration on the use of the Adaptive Optics Scanning Laser Ophthalmoscope (AOSLO). It first covers the background and the overall approaches to human AO retinal imaging, and the technology involved, and then concentrates on using AO retinal imaging to study the structure and function of the retina.

Enhanced retinal vasculature imaging with a rapidly configurable aperture.

In adaptive optics scanning laser ophthalmoscope (AOSLO) systems, capturing multiply scattered light can increase the contrast of the retinal microvasculature structure, cone inner segments, and retinal ganglion cells. Current systems generally use either a split detector or offset aperture approach to collect this light. We tested the ability of a spatial light modulator (SLM) as a rapidly configurable aperture to use more complex shapes to enhance the contrast of retinal structure. Particularly, we varied the orientation of a split detector aperture and explored the use of a more complex shape, the half annulus, to enhance the contrast of the retinal vasculature. We used the new approach to investigate the influence of scattering distance and orientation on vascular imaging.

Alterations to the Foveal Cone Mosaic of Diabetic Patients.

Purpose: We measured localized changes occurring in the foveal cone photoreceptors and related defects in the cone mosaic to alterations in the nearby retinal vasculature. Methods: The central 4° of the retina of 54 diabetic (53.7 ± 12.5 years) and 85 control (35.8 ± 15.2 years) participants were imaged with the Indiana adaptive optics scanning laser ophthalmoscope. Foveal cones and overlying retinal capillaries were imaged and infrared scanning laser ophthalmoscopy (IR SLO) images and optical coherence tomography (OCT) B-scans were obtained. Follow-up imaging sessions were performed with intervals from 4 to 50 months for 22 of the 54 diabetic participants. Results: The foveal cone mosaics of 49 of 54 diabetic participants were of sufficient quality to assess the absence or presence of small localized defects in the cone mosaic. In 13 of these 49 diabetic participants we found localized defects, visualized as sharp-edged areas of cones with diminished reflectivity. These small, localized areas ranged in size from 10 × 10 µm to 75 × 30 µm. Of these 13 participants with cone defects, 11 were imaged over periods from 4 to 50 months and the defects remained relatively stable. These dark regions were not shadows of overlying retinal vessels, but all participants with these localized defects had alterations in the juxtafoveal capillary network. Conclusions: The foveal cone mosaic can show localized areas of dark cones that persist over time, that apparently correspond to either missing or nonreflecting cones, and may be related to local retinal ischemia.

SD-OCT and Adaptive Optics Imaging of Outer Retinal Tubulation.

PURPOSE: To investigate outer retinal tubulation (ORT) using spectral domain optical coherence tomography (SD-OCT) and an adaptive optics scanning laser ophthalmoscope (AOSLO). To document the frequency of ORT in atrophic retinal conditions and quantify ORT dimensions versus adjacent retinal layers. METHODS: SD-OCT images were reviewed for the presence of retinal atrophy, scarring, and/or exudation. The greatest width of each ORT was quantified. Inner and outer retinal thicknesses adjacent to and within the area of ORT were measured for 18 patients. AOSLO imaged ORTs in five subjects with direct and scattered light imaging. RESULTS: ORT was identified in 47 of 76 subjects (61.8%) and in 65 eyes via SD-OCT in a wide range of conditions and ages, and in peripapillary atrophy. ORTs appeared as finger-like projections in atrophy, seen in the en face images. AOSLO showed some ORTs with bright cones that guide light within atrophic areas. Multiply scattered light mode AOSLO visualized variegated lines (18-35 µm) radiating from ORTs. The ORTs' width on OCT b-scan images varied from 70 to 509 µm. The inner retina at the ORT was significantly thinner than the adjacent retina, 135 vs.170 µm (P = .004), whereas the outer retina was significantly thicker, 115 vs. 80 µm (P = .03). CONCLUSIONS: ORTs are quite common in eyes with retinal atrophy in various disorders. ORTs demonstrate surviving photoreceptors in tubular structures found within otherwise nonsupportive atrophic areas that lack retinal pigment epithelium and choriocapillaris.