This PDF file contains the front matter associated with SPIE Proceedings Volume 10045, including the Title Page, Copyright information, Table of Contents, and Conference Committee listing.

UV-A/riboflavin collagen cross-linking (UV-CXL) is a clinical treatment for keratoconus that stiffens mechanically degraded corneal tissue. On the other hand, the intraocular pressure (IOP) can also affect the measured cornea elasticity. However, the combined effects of CXL at different IOPs on the corneal biomechanical properties are not well understood. In this work, the feasibility of assessing the viscoelasticity of the porcine cornea before and after CXL at various IOPs was investigated by using a noncontact method of optical coherence elastography (OCE) and a modified Lamb wave model. The modified wave model was first verified by comparison with finite element modeling, and then utilized to quantify the viscoelasticity of porcine corneas in the whole eye-globe configuration before and after CXL treatment at various IOPs. The results show that the elasticity of the cornea increased after CXL and that corneal stiffness was linear as a function of IOP. At IOPs of 15, 20, 25, and 30 mmHg, the relative increase in Young’s modulus after CXL was ~109%, ~86%, ~64%, and ~79%, respectively, while the shear viscosity decreased by ~86%, ~84%, ~83%, and ~81%. The modified Lamb wave model and OCE show promise for quantifying corneal viscoelasticity, which could provide a basis for customized CXL therapies and accurate disease detection.

Many ocular diseases such as glaucoma and uveitis can lead to the elevation of intraocular pressure (IOP). Previous research implies a link between elevated IOP and lens disease. However, the relationship between IOP elevation and biomechanical properties of the crystalline lens has not been directly studied yet. In this work, we investigated the biomechanical properties of porcine lens as a function of IOP by acoustic radiation force optical coherence elastography.

Diseases such as keratoconus can alter the orientation of collagen fibrils in the cornea. Moreover, therapeutic interventions such as UV-A/riboflavin corneal collagen crosslinking (CXL) can alter the collagen fibril arrangement. Therefore, the anisotropic characteristics of the cornea can provide vital information about tissue integrity. In this work, we utilize noncontact elastic wave imaging optical coherence elastography (EWI-OCE) to assess the elastic anisotropy and hysteresis of in situ porcine corneas as various intraocular pressures (IOP). In addition, we evaluated the effects of CXL on the mechanical anisotropy and hysteresis. OCE measurements were made at stepped meridional angles, and a sliding window algorithm spatially mapped the elasticity. A modified planar anisotropy coefficient was utilized to quantify the elastic anisotropy of the corneas. The results show that the stiffness and elastic anisotropy of the corneas were significantly affected by CXL and IOP (P<0.001), but the hysteresis was not significant (P<0.05). Moreover, the changes in elasticity due to CXL were angle-dependent (P<0.005). However, the changes in mechanical anisotropy from CXL were not angle-dependent (P>0.05).

Fungal keratitis can lead to pain and impaired vision. Current treatment options include antifungal agents and therapeutic penetrating keratoplasty. An emerging option for the management of keratitis is photodynamic antimicrobial therapy (PDAT) which uses a photosensitizer rose bengal activated with green light. Utilizing a pulsed irradiation, rather than the standard continuous irradiation may have a similar antimicrobial effect with less total energy. This study is to compare pulsed and continuous rose bengal mediated PDAT for inhibition of six fungal isolates on agar plates: Fusarium solani, Fusarium keratoplasticum, Aspergillus fumigatus, Candida albicans, Paecilomyces variotti, and Pseudoallescheria boydii. Isolates were mixed with 0.1% rose bengal and exposed to three irradiation conditions: (1) 30-minute continuous (10.8J/cm2), (2) 15-minute continuous (5.4J/cm2), (3) 30-minute pulsed (5.4J/cm2). Plates were photographed at 72 hours and analyzed with custom software. At 72 hours, 30-minute continuous rose bengal mediated PDAT inhibited all six fungal species. Fungal inhibition was analogous between 30-minute continuous and 30-minute pulsed test groups, with the exception of A. fumigatus. The 15-minute continuous irradiation was less effective when compared to both 30-minute continuous and 30-minute pulsed groups. These in vitro results demonstrate the potential strength of pulsed rose bengal mediated PDAT as an adjunct treatment modality for fungal keratitis.

Micropulse modulation in retinal laser therapy was intended to confine tissue heating around the light-absorbing layers, such as RPE and choroid, while the transparent retina is heated less as a result of slow heat diffusion. Current implementations use micropulses of 100-300μs at 500Hz, with overall pulse envelope of 100-300ms. The effect of such modulation compared to continuous-wave (CW) is not well characterized and misleading comparisons are made in the literature between exposures of different average power or overall duration. In this study, we modeled and measured the retinal tissue response to pulse trains with duty cycles from 4% (80μs pulse at 500Hz) to CW at overall envelope of 200ms and 20ms. Three thresholds of tissue response were measured in Dutch-belted rabbits: immediate (<3s after laser delivery) and delayed (1-5min) ophthalmoscopic visibility of lesions corresponding to photoreceptor damage, as well as fluorescein angiography visibility indicating RPE damage. Both the model and experimental results show that tissue response to micropulse modulation with long pulse envelope (200ms) is not significantly different from CW exposures at the same average power and duration. Heat confinement is improved with lower duty cycle (2%) and shorter pulse envelope (20ms), however further decrease in exposure duration raises the temperature dangerously close to vaporization. Pulse modulation cannot improve the therapeutic range of non-damaging thermal therapy since it is defined by the Arrhenius integral, regardless of the time course of hyperthermia. However, it does allow greater thermal stress to the RPE and underlying choroid while avoiding damage to neural retina.

Retinal and choroidal neovascularization play a pivotal role in the leading causes of blindness including macular degeneration and diabetic retinopathy (DR). Current therapy by focal laser photocoagulation can damage the normal surrounding cells, such as the photoreceptor inner and outer segments which are adjacent to the retinal pigment epithelium (RPE), due to the use of high laser energy and millisecond pulse duration. Therapies with pharmaceutical agents involve systemic administration of drugs, which can cause adverse effects and patients may become drug-resistant. We have developed a noninvasive photo-mediated ultrasound therapy (PUT) technique as a localized antivascular method, and applied it to remove micro blood vessels in the retina. PUT takes advantage of the high native optical contrast among biological tissues, and has the unique capability to self-target microvessels without causing unwanted damages to the surrounding tissues. This technique promotes cavitation activity in blood vessels by synergistically applying nanosecond laser pulses and ultrasound bursts. Through the interaction between cavitation and blood vessel wall, blood clots in microvessels and vasoconstriction can be induced. As a result, microvessels can be occluded. In comparison with other techniques that involves cavitation, both laser and ultrasound energy needed in PUT is significantly lower, and hence improves the safety in therapy.

We will present our results of evaluating the feasibility of using the mouse eye as a window for non-invasive, long-term, optical investigation of xenograft models, using multimodal, cellular-resolution ocular imaging. As an “approachable part of the brain”, the retina allows examination of such issues as drug delivery across the blood retinal barrier (BRB) and blood brain barrier (BBB). Our custom-built wide-field SLO/OCT provided repeatable in vivo imaging over many weeks, allowing quantitative tracking of tumor growth, the delivery of theranostic nanoparticles, and the measurement of tumor microenvironment responses. Additionally, we were able to specifically control the spatial extent of light activated photodynamic therapy (PDT) and photothermal therapy (PTT) via efficient free radical and heat generation at the tumor site, respectively.

Optical coherence tomography (OCT) has become standard in retinal imaging at the pre-clinical and clinical level by allowing non-invasive, three-dimensional imaging of the tissue structure. However, OCT lacks specificity to contrast agents that could be used for in vivo molecular imaging. We have performed in vivo photothermal optical coherence tomography (PT-OCT) of targeted gold nanorods in the mouse retina after the mice were injected systemically with the contrast agent. To our knowledge, we are the first to perform PT-OCT in the eye and image targeted gold nanorods with this technology. As a model of age-related wet macular degeneration, lesions were induced by laser photocoagulation in each mouse retina (n=12 eyes). Untargeted and targeted (anti-mouse CD102 antibody, labeling neovasculature) gold nanorods (peak absorption λ=750nm) were injected intravenously by tail-vein injection five days after lesion induction, and imaged the same day with PT-OCT. Our instrument is a spectral domain OCT system (λ=860nm) with a Titanium:Sapphire laser (λ=750nm) added to the beam path using a 50:50 coupler to heat the gold nanorods. We acquired PT-OCT volumes of one lesion per mouse eye. There was a significant increase in photothermal intensity per unit area of the lesion in the targeted gold nanorods group versus the saline control group and the untargeted gold nanorods group. This experiment demonstrates the feasibility of PT-OCT to image the distribution of molecular contrast agents in the mouse retina, including in highly scattering lesions. In the future we will use this method to identify new biomarkers linked with retinal disease.

We have recently reported observations of light-induced broadband fundus reflectance changes in two most commonly used strains of laboratory mice, C57Bl/6J (pigmented) and Balb/c (un- pigmented albino). The action spectrum of the reflectance increase corresponded to the absorption spectrum of mouse rhodopsin in situ. Spectral changes in mouse fundus reflectivity were calculated from measurements made by broadband spectrometer, interfaced with our mouse retinal SLO system, obtained before and after bleaching. This results were fitted with a model of mouse fundus reflectance, quantifying contributions from loss of rhodopsin absorption with bleaching, absorption by oxygenated hemoglobin (HbO2) in the choroid (Balb/c), and absorption by melanin (C57Bl/6J) additionally both mouse strains exhibited light-induced broadband reflectance changes explained as bleaching-induced reflectivity increases at photoreceptor inner segment/outer segment (IS/OS) junctions and OS tips. Here we present results investigating the kinetics of the increases in reflectivity with Optical Coherence Tomography operating in a 780-950 nm band.

To understand the pathogenesis of ophthalmic disease, utilizing small animal models such as mouse is necessary because of their ease of maintenance and availability. For identifying pathophysiology and drug development of retinal diseases in mouse model, optical coherence tomography angiography (OCTA) is promising imaging modality visualizing not only microstructure but also microvasculature. In this study, we serially imaged 3D structure and angiography of laser-induced choroidal neovascularization (CNV) in the mouse retina with/without anti-VEGF treatment. Also, the volume changes of CNV and avascular region in choroid layer are measured for identifying effects of anti-VEGF. A lab-built high-speed OCTA prototype using the wavelength-swept laser centered at 1040 nm with 230 kHz A-scan rate acquired 3-D volumetric data consisted of 1024 x 1024 x 3 A-scans. The OCTA scanned 1.7 mm x 1.7 mm area around ONH. For obtaining angiography, amplitude decorrelation from 3 consecutive B-scans at each position was generated. Seven days after the laser photocoagulation at mouse retina for generation of the laser-induced CNV, intravitreal administration of Fc and VEGF-Trap was given in the therapeutic arm. The OCTA were performed at 6, 14, 21 and 35 days after laser photocoagulation. Vasculatures of inner retina, outer retina and choroid layers were separately visualized after RPE flattening and layer segmentation. To investigate therapeutic effects of anti-VEGF treatment, the relative area and volume of CNV in outer retina layer is measured. Also, total volume of avascular zone surrounding the laser injury site in choroid layer is also analyzed.

Blood flow patterns and kinetics in the choriocapillaris are poorly understood owing to a lack of quantitative ophthalmic imaging techniques for studying microvascular flow in the eye. Compared with the proximal retinal vasculature, the more distal choroidal vasculature is relatively more challenging to probe. Magnetic Resonance Imaging and Doppler Ultrasound can assess the retina and choroid, but do not resolve the finer layers or microvasculature. While Optical Coherence Tomography (OCT) angiography produces high-quality choroidal images, attempts at quantification through Doppler-based methods have had mixed success. Here, we use a new technique called Dynamic Contrast OCT (DyC-OCT), which tracks the passage of an intravascular scattering contrast agent, to reveal laminar blood flow patterns in the retina and choroid in vivo. While conceptually similar to fluorescence angiography, DyC-OCT has the substantial benefit of depth resolution, which enables separation of retinal and choroidal microvasculature. The scattering contrast agent enables improved angiography of both macro- and microvasculature in the retina and choroid. Blood plasma transit times are measured in individual vessels, while flow and volume are quantified for each of the microvascular layers. As expected, the choriocapillaris had the highest volume and flow. Blood flow rates were estimated with an average retinal blood flow of 9.1 ± 4.3 μL/min and an average choroidal blood flow of 40 ± 18.3 μL/min in the rat eye. These rates are consistent with previous literature. DyC-OCT affords a new perspective on the poorly understood choriocapillaris blood flow and kinetics and may be useful for studying outer retinal diseases.

Visually evoked changes of retinal blood flow can serve as an important research tool to investigate eye disease such as glaucoma and diabetic retinopathy. In this study we used a combined, research-grade, high-resolution Doppler OCT+ERG system to study changes in the retinal blood flow (RBF) and retinal neuronal activity in response to visual stimuli of different intensities, durations and type (flicker vs single flash). Specifically, we used white light stimuli of 10 ms and 200 ms single flash, 1s and 2s for flickers stimuli of 20% duty cycle. The study was conducted in-vivo in pigmented rats. Both single flash (SF) and flicker stimuli caused increase in the RBF. The 10 ms SF stimulus did not generate any consistent measurable response, while the 200 ms SF of the same intensity generated ~4% change in the RBF peaking at ~1.5 s after the stimulus onset. Single flash stimuli introduced ~2x smaller change in RBF and ~30% earlier RBF peak response compared to flicker stimuli of the same intensity and duration. Doubling the intensity of SF or flicker stimuli increased the RBF peak magnitude by ~1.5x. Shortening the flicker stimulus duration by 2x increased the RBF recovery rate by 2x, however, had no effect on the rate of RBF change from baseline to peak.

Zebrafish have been identified as an ideal model for angiogenesis because of anatomical and functional similarities with other vertebrates. The scale and complexity of zebrafish assays are limited by the need to manually treat and serially screen animals, and recent technological advances have focused on automation and improving throughput. Here, we use optical coherence tomography (OCT) and OCT angiography (OCT-A) to perform noninvasive, in vivo imaging of retinal vasculature in zebrafish. OCT-A summed voxel projections were low pass filtered and skeletonized to create an en face vascular map prior to connectivity analysis. Vascular segmentation was referenced to the optic nerve head (ONH), which was identified by automatically segmenting the retinal pigment epithelium boundary on the OCT structural volume. The first vessel branch generation was identified as skeleton segments with branch points closest to the ONH, and subsequent generations were found iteratively by expanding the search space outwards from the ONH. Biometric parameters, including length, curvature, and branch angle of each vessel segment were calculated and grouped by branch generation. Despite manual handling and alignment of each animal over multiple time points, we observe distinct qualitative patterns that enable unique identification of each eye from individual animals. We believe this OCT-based retinal biometry method can be applied for automated animal identification and handling in high-throughput organism-level pharmacological assays and genetic screens. In addition, these extracted features may enable high-resolution quantification of longitudinal vascular changes as a method for studying zebrafish models of retinal neovascularization and vascular remodeling.

Keratoconus causes progressive morphological changes in the corneal epithelium (EPI), Bowman’s membrane (BM) and anterior stroma. However, it is still not well understood if KC originates in the corneal epithelium and propagates to the anterior stroma through disruptions of the BM, or vice versa. In this study we used a sub-micrometer axial resolution OCT system to image in-vivo the cellular structure of the EPI layer and the fibrous structure of the BM and the anterior stroma in mild to advanced keratoconics, as well as healthy subjects. The imaging study was approved by the University of Waterloo Human Research Ethics Committee. The OCT system operates in the 800 nm spectral region at 34 kHz image acquisition rate and provides 0.95 um axial and < 2 um lateral resolution in corneal tissue, which is sufficient to visualize the cellular structure of the corneal epithelium and the fibrous structure of the BM. In some subjects, localized thinning and thickening of the EPI layer was observed, while there was no visible damage to the BM or anterior stroma. In other subjects, localized breakage of the stromal collagen fibrils was observed with no significant morphological changes of the corneal EPI.

Keratoconus is an eye disorder that features a reduced stiffness of the cornea and its consequent pathological deformation. Cross-Linking (CXL) treatment has proven useful in hindering the progression of keratoconus, offering a minimally-invasive alternative to corneal surgical transplantation. In this study, the biomechanical characteristics of a human keratoconic cornea were clinically examined in vivo soon before keratoplasty, and the morphological alterations of the collagen scaffold in the same cornea were examined ex vivo by means of Second-Harmonic Generation (SHG) microscopy. A healthy cornea and a CXL-treated keratoconus were compared. In particular, the lamellar organization in the three corneal samples was characterized in different stromal layers by detecting both forward- and backwardscattered SHG signal and then considering the forward/backward (F/B) ratio as parameter. The F/B ratio was used to characterize the morphological organization of collagen lamellae within different stromal layers, finding an increased disorder at the level of Bowman's membrane, opposed to a more regular organization within deeper stromal layers in all the examined samples. The organization of collagen lamellae in CXL-treated keratoconic samples was similar to that one found in healthy corneas, demonstrating that the CXL is able to rearrange the collagen scaffold and partially recover the properties of a healthy condition. The obtained results are in agreement with previous results obtained in studies aimed at monitoring the organization of fibrillar collagen using F/B SHG ratio. In conclusion, the proposed method might be useful for both diagnosing keratoconus as well as for monitoring the effects of the CXL treatment.

Full-Field Optical Coherence Tomography (FF-OCT) reveals submicrometric morphological details in retinal explants without the use of contrast agents. Notably, in the nerve fiber and ganglion cell layers, FF-OCT images reveal nerve fibers bundles, single axons, capillaries and even some ganglion cell bodies. Dynamic FF-OCT (D-FF-OCT) takes advantage of the temporal evolution of the local FF-OCT signal to reveal a movement-dependent contrast inside tissues. Notably, the D-FF-OCT signal depends on cellular motility and membrane fluctuations. Compared to regular FF-OCT images, the signal from stationary structures such as nerve fibers is reduced, and contrast inside cells is enhanced, revealing many more cells, as well as the position of nuclei, and cell metabolism. We used a multimodal D-FF-OCT and fluorescence microscope to compare and identify the structures observed in both FF-OCT and D-FF-OCT. In the ganglion cell and inner nuclear layers in both macaque and mouse, two different cell sizes could be measured, which correlated well with ganglion and amacrine cell diameters found in the literature for these two species. We could also detect cell bodies of the photoreceptors in the outer nuclear layer. To our knowledge, this is the first time that an OCT technique can reveal these cell bodies. Finally, to investigate post-mortem tissue changes, time series were acquired over periods of 24 hours and cell contrast was plotted in time to monitor the decrease in intracellular activity over time. It is anticipated that dynamic FF-OCT may be used to non-invasively monitor viability and functional changes in the retina.

Glaucoma is a progressive optic neuropathy, characterized by the selective loss of retinal ganglion cells (RGCs). Therefore, monitoring the change of number or morphology of RGC is essential for the early detection as well as investigation of pathophysiology of glaucoma. Since RGC layer is transparent and hyporeflective, the direct optical visualization of RGCs has not been successful so far. Therefore, glaucoma evaluation mostly depends on indirect diagnostic methods such as the evaluation of optic disc morphology or retinal nerve fiber layer thickness measurement by optical coherence tomography. We have previously demonstrated single photoreceptor cell imaging with differential interference contrast (DIC) microscopy. Herein, we successfully visualized single RGC using DIC microscopy. Since RGC layer is much less reflective than photoreceptor layer, various techniques including the control of light wavelength and bandwidth using a tunable band pass filter were adopted to reduce the chromatic aberration in z-axis for higher and clearer resolution. To verify that the imaged cells were the RGCs, the flat-mounted retina of Sprague-Dawley rat, in which the RGCs were retrogradely labeled with fluorescence, was observed by both fluorescence and DIC microscopies for direct comparison. We have confirmed that the cell images obtained by fluorescence microscopy were perfectly matched with cell images by DIC microscopy. As conclusion, we have visualized single RGC with DIC microscopy, and confirmed with fluorescence microscopy.

We evaluated intraretinal RPE migration in AMD using multimodal imaging including polarimetric images. Depolarized light images were computed using a PS-SLO. M-DOPU was calculated using multifunctional Jones-matrix OCT. RPE migration was detected in 59 of 155 eyes. Focal similarities could be confirmed among en-face projection images of minimum M-DOPU, depolarized light images, and NIR-AF images.

Purpose: To evaluate the effects of four anatomical parameters (angle between superior and inferior temporal retinal arteries [inter-artery angle, IAA], optic disc [OD] rotation, retinal curvature, and central retinal vessel trunk entry point location [CRVTL]) on retinal nerve fiber layer thickness (RNFLT) abnormality marks by OCT machines. Methods: Cirrus OCT circumpapillary RNFLT measurements and Humphrey visual fields (HVF 24-2) of 421 patients from a large glaucoma clinic were included. Ellipses were fitted to the OD borders. Ellipse rotation relative to the vertical axis defined OD rotation. CRVTL was manually marked on the horizontal axis of the ellipse on the OCT fundus image. IAA was calculated between manually marked retinal artery locations at the 1.73mm radius around OD. Retinal curvature was determined by the inner limiting membrane on the horizontal B-scan closest to the OD center. For each location on the circumpapillary scanning area, logistic regression was used to determine if each of the four parameters had a significant impact on RNFLT abnormality marks independent of disease severity. The results are presented on spatial maps of the entire scanning area. Results: Variations in IAA significantly influenced abnormality marks on 38.8% of the total scanning area, followed by CRVTL (19.2%) and retinal curvature (18.7%). The effect of OD rotation was negligible (<1%). Conclusions: A natural variation in IAA, retinal curvature, and CRVTL can affect OCT abnormality ratings, which may bias clinical diagnosis. Our spatial maps may help OCT manufacturers to introduce location specific norms to ensure that abnormality marks indicate ocular disease instead of variations in eye anatomy.

The present study aimed to elucidate if comparison of angular segments of Pigment epithelium central limit- Inner limit of the retina Minimal Distance, measured over 2π radians in the frontal plane (PIMD-2π) between visits of a patient, renders sufficient precision for detection of loss of nerve fibers in the optic nerve head. An optic nerve head raster scanned cube was captured with a TOPCON 3D OCT 2000 (Topcon, Japan) device in one early to moderate stage glaucoma eye of each of 13 patients. All eyes were recorded at two visits less than 1 month apart. At each visit, 3 volumes were captured. Each volume was extracted from the OCT device for analysis. Then, angular PIMD was segmented three times over 2π radians in the frontal plane, resolved with a semi-automatic algorithm in 500 equally separated steps, PIMD-2π. It was found that individual segmentations within volumes, within visits, within subjects can be phase adjusted to each other in the frontal plane using cross-correlation. Cross correlation was also used to phase adjust volumes within visits within subjects and visits to each other within subjects. Then, PIMD-2π for each subject was split into 250 bundles of 2 adjacent PIMDs. Finally, the sources of variation for estimates of segments of PIMD-2π were derived with analysis of variance assuming a mixed model. The variation among adjacent PIMDS was found very small in relation to the variation among segmentations. The variation among visits was found insignificant in relation to the variation among volumes and the variance for segmentations was found to be on the order of 20 % of that for volumes. The estimated variances imply that, if 3 segmentations are averaged within a volume and at least 10 volumes are averaged within a visit, it is possible to estimate around a 10 % reduction of a PIMD-2π segment from baseline to a subsequent visit as significant. Considering a loss rate for a PIMD-2π segment of 23 μm/yr., 4 visits per year, and averaging 3 segmentations per volume and 3 volumes per visit, a significant reduction from baseline can be detected with a power of 80 % in about 18 months. At higher loss rate for a PIMD-2π segment, a significant difference from baseline can be detected earlier. Averaging over more volumes per visit considerably decreases the time for detection of a significant reduction of a segment of PIMD-2π. Increasing the number of segmentations averaged per visit only slightly reduces the time for detection of a significant reduction. It is concluded that phase adjustment in the frontal plane with cross correlation allows high precision estimates of a segment of PIMD-2π that imply substantially shorter followup time for detection of a significant change than mean deviation (MD) in a visual field estimated with the Humphrey perimeter or neural rim area (NRA) estimated with the Heidelberg retinal tomograph.

Glaucoma is a group of eye diseases which results in optic nerve damage and vision loss. Optical coherence tomography (OCT) has been widely used to investigate geometric risk factor of glaucoma. However, material properties of ONH are also important to understand intra-ocular pressure related stress. We developed Jones-matrix based multifunctional posterior eye OCT (JM-OCT), which uses 1-μm band swept-source with a 100-kHz A-line rate. It provides three different optical properties, attenuation coefficient (AC), local birefringence (LB), and optical coherence angiography (OCA). We investigated the utility those properties for the investigation of normal ONH cases. 3 mm x 3 mm area around ONH was scanned for each eye, and biometric parameters were measured in hospital. Statistical analyses were performed with the mean values of above parameters at the regions of prelamina, lamina cribrosa, peripapillary sclera, and peripapillary nerve fiber layer, and biometric parameters of age, axial eye length, refractive error, and intraocular pressure. In qualitative observation, the lamina cribrosa generally shows more hyper signals in AC, LB, and OCA than prelamina. In t-test, AC, LB, and OCA showed significant difference (p < 0.05) between prelamina and lamina cribrosa, while conventional OCT did not. In correlation test, axial eye length is positively correlated with LB and AC in lamina cribrosa. And these LB and AC are also negatively correlated with the refractive error. Age was found to be negatively correlated with OCA in lamina cribrosa.

BACKGROUND: Vitreous opacities and posterior vitreous detachment (PVD) disturb vision by degrading contrast sensitivity (AJO 172:7-12, 2016). Increased light scattering is the presumed mechanism. To test this hypothesis, dynamic light scattering (DLS) was performed on excised vitreous of patients with clinically significant floaters, and compared to macular pucker controls. METHODS: Undiluted, unfixed vitreous was procured during 25-gauge vitrectomy in 14 subjects (age = 59 ± 6.6 years) with clinically significant vitreous floaters, and 6 controls (age = 66.5 ± 8.7 years; P = 0.10) with macular pucker. Total protein concentration was determined by fluorescent Quant-iT^TM protein assay kit (Invitrogen/Molecular Probes, Eugene, OR) with bovine serum albumin (0500 ng/ml) as a standard. Fluorescence (excitation at 470 nm and emission at 570 nm) was measured using a Gemini XPS Dual-Scanning Microplate Spectrofluorometer and data analyzed using SoftMax Pro software (Molecular Devices, Sunnyvale, CA). DLS (NS300, Malvern Instruments, Westborough, MA) measurements were performed in each specimen after 10-fold dilution in phosphate buffered saline to optimize concentration in each specimen and determine the mean number of particles, the particle size distributions, and the average particle sizes. RESULTS: Total protein concentration in vitreous specimens trended higher in macular pucker controls (1037 ± 1038 μg/mL) than eyes with vitreous floaters (353.7 ± 141.1 μg/mL; P = 0.08). When normalized to total protein concentration, the number of particles in vitreous from floater eyes was more than 2-fold greater than controls (P < 0.04). Particle size distributions were similarly two-fold greater in vitreous from floater subjects as compared to controls (P < 0.05). The average particle size in vitreous from floater eyes was 315.8 ± 194.6 nm, compared to 147.7 ± 129.3 nm in macular pucker controls (P = 0.039). CONCLUSIONS: Vitreous from eyes with clinically significant floaters contains more particles of larger sizes as compared to controls, likely accounting for the degradation of contrast sensitivity previously found in these patients (Retina 34:1062-8, 2014; IOVS 56:1611–7, 2015; AJO 172:7-12, 2016). DLS could elucidate the underlying molecular abnormalities in patients afflicted with bothersome vitreous floaters and help develop clinical tools to better measure vitreous floaters as well as test the efficacy of various therapies.

A motion - corrected en face optical coherence tomography angiography (OCT - A) imaging method is presented in this paper. This method was designed to correct eye motion artifacts in en face OCT - A images automatically using a software. A modified Lissajous scanning pattern, which is compatible with OCT - A, was adopted as the scanning protocol for the optical coherence tomography (OCT) machine to obtain the OCT - A data. The OCT - A data was then processed using motion correction algorithm that was tailored for the modified Lissajous scanning pattern to correct the eye motion artifacts in the en face plane. The OCT - A slab between the inner limiting membrane and outer plexiform layer was segmented to provide an enhanced motion - corrected en face OCT - A image of the inner retinal flow. The comparison of the en face OCT - A images with and without motion correction show that our motion correction algorithm corrected the eye motion artifacts effectively. A motion - corrected en face OCT - A image of target layer is presented to confirm our method.

To investigate the application of wide field OCT angiography (OCTA) in living human eye. Normal and pathologic eyes were recruited and imaged by a 1060 nm swept source OCTA system with A-line speed of 100 kHz provided by Carl Zeiss Meditec. Inc.. Wide field OCTA images were generated in a single scan within 5 seconds based on the tracking capability installed in the system with 9 x 9 mm² and 12 x 12 mm² field of view and sampled by 500 A-lines x 500 Bframes with 2 repetitions in the same location for one 3D data. Complex optical microangiography (OMAG) algorithm was used to extract the blood flow information. The en face maximum projection provided by the device was used to generate 2-dimensional angiograms of different layers and color-code images. Wide field en face OCTA images of different macular diseases showed a great agreement with fluorescein angiography (FA). Meanwhile, OCTA provides depth-resolved information and detailed vascular images of venous occlusion and DR patients in far peripheral region, and choroidal vessels imaging in serpiginous choroidopathy patient, providing a better visualization of vascular network compared to FA.

Using a commercial available 200K swept source laser, we demonstrated high resolution wide field angiographic imaging of human retinal. 8mm by 8mm and 10mm by 6mm retina areas were imaged in a single scan within 4 seconds. By montaging four 10 x 6mm scan, 10 x 20mm wide field OCT angiography images were demonstrated.

Optical coherence tomography (OCT) allows for micron scale imaging of the human retina and cornea. Previous research and commercial intraoperative OCT prototypes have been limited to live B-scan imaging because they were based on previous-generation spectral domain OCT systems. Our group has developed and reported on an intraoperative microscope integrated OCT system based on a 100 kHz commercial swept source laser. This system is capable of live 4D imaging, and with a heads up display allows for dynamic intraoperative visualization of retinal structures, tool tissue interaction, and surgical maneuvers. OCT angiography (OCTA) is an emerging OCT technology that allows for imaging of retinal vasculature without the use of potentially harmful contrast agents. This structural information can provide insights into the state and development of a wide range of ophthalmic pathologies. The addition of OCTA into intraoperative OCT could allow for monitoring of changes in retinal vasculature during surgery and imaging of traditionally non-compliant patients. In this work we provide a brief update of intraoperative 4D MIOCT across a range of pathologies, and demonstrate intraoperative OCTA for the first time. To the best of knowledge, this is the first report of intraoperative OCTA, as well as the first OCTA images ever acquired in an infant.

We demonstrate OCT angiography (OCTA) and Doppler OCT imaging of the choroid in the eyes of two healthy volunteers and in a geographic atrophy case. We show that visualization of specific choroidal layers requires selection of appropriate OCTA methods. We investigate how imaging speed, B-scan averaging and scanning density influence visualization of various choroidal vessels. We introduce spatial power spectrum analysis of OCT en face angiographic projections as a method of quantitative analysis of choroicapillaris morphology. We explore the possibility of Doppler OCT imaging to provide information about directionality of blood flow in choroidal vessels. To achieve these goals, we have developed OCT systems utilizing an FDML laser operating at 1.7 MHz sweep rate, at 1060 nm center wavelength, and with 7.5 μm axial imaging resolution. A correlation mapping OCA method was implemented for visualization of the vessels. Joint Spectral and Time domain OCT (STdOCT) technique was used for Doppler OCT imaging.

Adaptive optics optical coherence tomography (AO-OCT) systems capable of 3D high resolution imaging have been applied to posterior eye imaging in order to resolve the fine morphological features in the retina. Human cone photoreceptors have been extensively imaged and studied for the investigation of retinal degeneration resulting in photoreceptor cell death. However, there are still limitations of conventional approaches to AO in the clinic, such as relatively small field-of-view (FOV) and the complexities in system design and operation. In this research, a recently developed phase-resolved Sensorless AO Swept Source based OCT (SAO-SS-OCT) system which is compact in size and easy to operate is presented. Owing to its lens-based system design, wide-field imaging can be performed up to 6° on the retina. A phase stabilization unit was integrated with the OCT system. With the phase stabilized OCT signal, we constructed retinal micro-vasculature image using a phase variance technique. The retinal vasculature image was used to align and average multiple OCT volumes acquired sequentially. The contrast-enhanced photoreceptor projection image was then extracted from the averaged volume, and analyzed based on its morphological features through a novel photoreceptor structure evaluation algorithm. The retinas of twelve human research subjects (10 normal and 2 pathological cases) were measured in vivo. Quantitative parameters used for evaluating the cone photoreceptor mosaic such as cell density, cell area, and mosaic regularity are presented and discussed. The SAO-SS-OCT system and the proposed photoreceptor evaluation method has significant potential to reveal early stage retinal diseases associated with retinal degeneration.

Retinal pigment epithelium (RPE) cells are vital to health of the outer retina, however, are often compromised in ageing and ocular diseases that lead to blindness. Early manifestation of RPE disruption occurs at the cellular level, but while in vivo biomarkers at this scale hold considerable promise, RPE cells have proven extremely challenging to image in the living human eye. Recently we addressed this problem by using organelle motility as a novel contrast agent to enhance the RPE cell in conjunction with 3D resolution of adaptive optics-optical coherence tomography (AO-OCT) to section the RPE layer. In this study, we expand on the central novelty of our method – organelle motility – by characterizing the dynamics of the motility in individual RPE cells, important because of its direct link to RPE physiology. To do this, AO-OCT videos of the same retinal patch were acquired at approximately 1 min intervals or less, time stamped, and registered in 3D with sub-cellular accuracy. Motility was quantified by an exponential decay time constant, the time for motility to decorrelate the speckle field across an RPE cell. In two normal subjects, we found the decay time constant to be just 3 seconds, thus indicating rapid motility in normal RPE cells.

Combined adaptive optics (AO) optical coherence tomography (OCT) scanning laser ophthalmoscopy (SLO) imaging allows simultaneous en face and cross sectional views of the retina. We describe improvements to our AO-OCT-SLO system and highlight its resolution capability and clinical utility by presenting results from 3 control and 4 dry agerelated macular degeneration (AMD) subjects. From a group of subjects with healthy eyes, OCT A-scans were grouped as originating from cones or rods and were averaged. The resulting reflectance profiles were then used to identify the location of cone and rod segments. Results for rods and cones were compared, with the focus on inner segment (IS) and outer segment (OS) structures and where these cells embed into the retinal pigment epithelium (RPE). In the AMD patients, cone IS and OS lengths were measured over and around drusen for two retinal regions (fovea–2° and 2°–4°), and those results were correlated to drusen height. For the fovea–2° region, the drusen height that caused statistically significant shortening of cone ISL and OSL compared to the unaffected adjacent area were 40 μm and 50 μm respectively (p = 0.009, and p < 0.001, respectively). For the 2°–4° region, the equivalent drusen heights that caused significant shortening of segment length were 60 μm for IS (p = 0.017) and 80 μm for OS (p < 0.001)

Absorption of light by photoreceptors initiates vision, but also leads to accumulation of toxic photo-oxidative compounds in the photoreceptor outer segment (OS). To prevent this buildup, small packets of OS discs are periodically pruned from the distal end of the OS, a process called disc shedding. Unfortunately dysfunction in any part of the shedding event can lead to photoreceptor and RPE dystrophy, and has been implicated in numerous retinal diseases, including age related macular degeneration and retinitis pigmentosa. While much is known about the complex molecular and signaling pathways that underpin shedding, all of these advancements have occurred in animal models using postmortem eyes. How these translate to the living retina and to humans remain major obstacles. To that end, we have recently discovered the optical signature of cone OS disc shedding in the living human retina, measured noninvasively using optical coherence tomography equipped with adaptive optics in conjunction with post processing methods to track and monitor individual cones in 4D. In this study, we improve on this method in several key areas: increasing image acquisition up to MHz A-scan rates, improving reliability to detect disc shedding events, establishing system precision, and developing cone tracking for use across the entire awake cycle. Thousands of cones were successfully imaged and tracked over the 17 hour period in two healthy subjects. Shedding events were detected in 79.5% and 77.4% of the tracked cones. Similar to previous animal studies, shedding prevalence exhibited a diurnal rhythm. But we were surprised to find that for these two subjects shedding occurred across the entire day with broad, elevated frequency in the morning and decreasing frequency as the day progressed. Consistent with this, traces of the average cone OS length revealed shedding dominated in the morning and afternoon and renewal in the evening.

In this report, we demonstrate that in a coherence revival (CR) based swept source optical coherence tomography (SSOCT) set-up, real-time cross-sectional long-range images can be produced via the Master Slave (MS) method. The total tolerance of the MS method to nonlinear tuning, to dispersion in the interferometer and to dispersion due to the laser cavity, makes the MS ideally suited to the practice of coherence revival. In addition, enhanced versatility is allowed by the MS method in displaying shorter axial range images than that determined by the digital sampling of the data. This brings an immediate improvement in the speed of displaying cross-sectional images at high rates without the need of extra hardware such as graphics processing units or field programmable gate arrays. The long axial range of the coherence revival regime is proven with images of the anterior segment of healthy human eye.

According to the World Health Organization (WHO), corneal diseases alongside with cataract and retinal diseases are major causes of blindness worldwide. For the 95.5% of corneal blindness cases, prevention or rehabilitation could have been possible without negative consequences for vision, provided that disease is diagnosed early. However, diagnostics at the early stage requires cellular-level resolution, which is not achieved with routinely used Slit-lamp and OCT instruments. Confocal microscopy allows examination of the cornea at a resolution approaching histological detail, however requires contact with a patient’s eye. The recently developed full-field OCT technique, in which 2D en face tangential optical slices are directly recorded on a camera, was successfully applied for ex vivo eye imaging. However, in vivo human eye imaging has not been demonstrated yet. Here we present a novel non-contact full-field OCT system, which is capable of imaging in air and, therefore, shows potential for in vivo cornea imaging in patients. The first cellular-level resolution ex vivo images of cornea, obtained in a completely non-contact way, were demonstrated. We were able to scan through the entire cornea (400 µm) and resolve epithelium, Bowman’s layer, stroma and endothelium. FFOCT images of the human cornea in vivo were obtained for the first time. The epithelium structures and stromal keratocyte cells were distinguishable. Both ex vivo and in vivo images were acquired with a large (1.26 mm x 1.26 mm) field of view. Cellular details in obtained images make this device a promising candidate for realization of high-resolution in vivo cornea imaging.

Optical coherence tomography (OCT) has revolutionized clinical observation of the eye and is an indispensable part of the modern ophthalmic practice. Unlike many other ophthalmic imaging techniques, OCT provides three-dimensional information about the imaged eye. However, conventional clinical OCT systems image only the anterior or the posterior eye during a single acquisition. Newer OCT systems have begun to image both during the same acquisition but with compromises such as limited field of view in the posterior eye or requiring rapid switching between the anterior and posterior eye during the scan. We describe here the development and demonstration of an OCT system with truly simultaneous imaging of both the anterior and posterior eye capable of imaging the full anterior chamber width and 50° on the retina (macula, optic nerve, and arcades). The whole eye OCT system was developed using custom optics and optomechanics. Polarization was utilized to separate the imaging channels. We utilized a 200kHz swept-source laser (Axsun Technologies) centered at 1040±50nm of bandwidth. The clock signal generated by the laser was interpolated 4x to generate 5504 samples per laser sweep. With the whole eye OCT system, we simultaneously acquired anterior and posterior segments with repeated B-scans as well as three-dimensional volumes from seven healthy volunteers (other than refractive error). On three of these volunteers, whole eye OCT and partial coherence interferometry (LenStar PCI, Haag-Streit) were used to measure axial eye length. We measured a mean repeatability of ±47µm with whole eye OCT and a mean difference from PCI of -68µm.

Conventional scanning laser ophthalmoscopy (SLO) utilizes a finite collection pinhole at a retinal conjugate plane to strongly reject out-of-focus light while primarily transmitting the in-focus, retinal backscattered signal. However, to improve lateral resolution, a sub-Airy disk collection pinhole is necessary, which drastically reduces the signal-to-noise ratio (SNR) of the system and is thus not commonly employed. Recently, an all-optical, super-resolution microscopy technique known as optical photon reassignment (OPRA) microscopy (also known as re-scan confocal microscopy) has been developed to bypass this fundamental tradeoff between resolution and SNR in confocal microscopy. We present a methodology and system design for obtaining super resolution in retinal imaging by combining the concepts of SLO and OPRA microscopy. The resolution improvement of the system was quantified using a 1951 USAF target at a telecentric intermediate image plane. Retinal images from human volunteers were acquired with this system both with and without using the OPRA technique to demonstrate the resolution improvement when imaging parafoveal cone photoreceptors. Finally, we quantified the resolution improvement in the retina by analyzing the radially averaged power spectrum of the retinal images.

Scanning laser ophthalmoscopy (SLO) and optical coherence tomography (OCT) enable noninvasive in vivo diagnostic imaging and provide complementary en face and depth-resolved visualization of ophthalmic structures, respectively. We previously demonstrated concurrent multimodal swept-source spectrally encoded scanning laser ophthalmoscopy and OCT (SS-SESLO-OCT) at 1060 nm using a swept-source and double clad fiber coupler. Here, we present system enhancements and novel designs for a modular SS-SESLO-OCT scan-head that can be coupled to ophthalmic surgical microscope-integrated and slit-lamp imaging optics. Multimodal SS-SESLO-OCT was demonstrated using a custom-built swept-source OCT engine with a 200 kHz 1060 nm source that was optically buffered for concurrent SESLO and OCT imaging at 100% duty cycle and 400 kHz sweep-rate. A shared optical relay and fast-axis galvanometer ensured inherent co-registration between SESLO and OCT field-of-views and concurrent acquisition of an en face SESLO image with each OCT cross-section. SESLO and OCT frames were acquired at 200 fps with 2560 x 2000 pix. (spectral x lateral). We show in vivo human ophthalmic imaging data using surgical microscope-integrated and slit-lamp imaging relays to demonstrate the utility of our SS-SESLO-OCT design. Our self-contained modular scan-head can be used for either intraoperative guidance or clinical diagnostics and reduces the complexity, cost, and maintenance required for clinical translation of these technologies. We believe concurrent multimodal SS-SESLO-OCT may benefit 1) intraoperative imaging by allowing for real-time surgical feedback, instrument tracking, and overlays of computationally extracted image-based surrogate biomarkers of disease, and 2) slit-lamp imaging by enabling aiming, image registration, and multi-field mosaicking.

Age-related changes in the crystalline lens shape and refractive index gradient produce changes in dioptric power and high-order aberrations that influence the optics of the whole eye and contribute to a decrease in overall visual quality. Despite their key role, the changes in lens shape and refractive index gradient with age and accommodation and their effects on high-order aberrations are still not well understood. The goal of this project was to develop a combined laser ray tracing (LRT) and optical coherence tomography (OCT) system to measure high-order aberrations, shape and refractive index gradient in non-human primate and human lenses. A miniature motorized lens stretching system was built to enable imaging and aberrometry of the lens during simulated accommodation. A positioning system was also built to enable on- and off-axis OCT imaging and aberrometry for characterization of the peripheral defocus of the lens. We demonstrated the capability of the LRT-OCT system to produce OCT images and aberration measurements of crystalline lens with age and accommodation in vitro. In future work, the information acquired with the LRT-OCT system will be used to develop an accurate age-dependent lens model to predict the role of the lens in the development of refractive error and aberrations of the whole eye.

Purpose: To determine the dynamic interaction between ciliary muscle and lens during accommodation and disaccommodation through synchronous imaging of ciliary muscle and lens response to pulse stimulus Methods: The ciliary muscle and lens were imaged simultaneously in a 33 year old subject responding to a 4D pulse stimulus (accommodative stimulus at 1.7 s, disaccommodative stimulus at 7.7 s) using an existing imaging system (Ruggeri et al, 2016) consisting of an Anterior Segment Optical Coherence Tomography system, Ciliary Muscle Optical Coherence Tomography system, and custom-built accommodation module. OCT images were recorded at an effective frame rate of 13.0 frames per second for a total scan time of 11.5 s. An automated segmentation algorithm was applied to images of the anterior segment to detect the boundaries of the cornea and lens, from which lens thickness was extracted. Segmentation of the ciliary muscle was performed manually and then corrected for distortion due to refraction of the beam to obtain measurements of thicknesses at the apex and fixed distances from the scleral spur. Results: The dynamic biometric response to a pulse stimulus at 4D was determined for both the ciliary muscle and lens, suggesting the ciliary muscle and lens interact differently in accommodation and disaccommodation. Conclusions: The study introduces new data and analyses of the ciliary muscle and lens interaction during a complete accommodative response from the relaxed to the accommodated state and back, providing insight into the interplay between individual elements in the accommodative system and how their relationships may change with age.

For ametropic eyes, LASIK is a common surgical procedure to correct the refractive error. However, the correction of hyperopia is more difficult than that of myopia because the increase of the central corneal curvature by excimer ablation is only possible by intrastromal tissue removal within a ring-like zone in the corneal periphery. For high hyperopia, the ring-shaped indentation leads to problems with the stability and reproducibility of the correction due to epithelial regrowth. Recently, it was shown that the correction of hyperopia can be achieved by implanting intracorneal inlays into a laser-dissected intrastromal pocket. In this paper we demonstrate the feasibility of a new approach in which a transparent, and biocompatible liquid filler material is injected into a laser-dissected corneal pocket, and the refractive change is monitored via OCT. This technique allows for a precise and adjustable change of the corneal curvature. Precise cutting of the intrastromal pocket was achieved by focusing UV laser picosecond pulses from a microchip laser system into the cornea. After laser dissection, the transparent filler material was injected into the pocket. The increase of the refractive power by filler injection was evaluated by taking OCT images from the cornea. With this novel technique, it is possible to precisely correct hyperopia of up to 10 diopters. An astigmatism correction is also possible by using ellipsoidal intrastromal pockets.

Glaucoma is the leading cause of irreversible blindness worldwide. Due to its wide prevalence, effective screening tools are necessary. The purpose of this project is to design and evaluate a system that enables portable, cost effective, smartphone based visual field screening based on frequency doubling technology. The system is comprised of an Android smartphone to display frequency doubling stimuli and handle processing, a Bluetooth remote for user input, and a virtual reality headset to simulate the exam. The LG Nexus 5 smartphone and BoboVR Z3 virtual reality headset were used for their screen size and lens configuration, respectively. The system is capable of running the C-20, N-30, 24-2, and 30-2 testing patterns. Unlike the existing system, the smartphone FDT tests both eyes concurrently by showing the same background to both eyes but only displaying the stimulus to one eye at a time. Both the Humphrey Zeiss FDT and the smartphone FDT were tested on five subjects without a history of ocular disease with the C-20 testing pattern. The smartphone FDT successfully produced frequency doubling stimuli at the correct spatial and temporal frequency. Subjects could not tell which eye was being tested. All five subjects preferred the smartphone FDT to the Humphrey Zeiss FDT due to comfort and ease of use. The smartphone FDT is a low-cost, portable visual field screening device that can be used as a screening tool for glaucoma.

Purpose: Objective pupilloperimetry in healthy subjects and patients with Best’s Vitelliform Macular Dystrophy using a chromatic multifocal pupillometer (CMP). Methods: A CMP (Accutome Inc) was used to record pupillary responses (PR) of 17 healthy subjects and 5 Best’s patients. Red and blue light stimuli were presented at 76 locations of a 16.2 degree VF. The PR of patients were compared with their findings on Humphrey's 24-2 perimetry, Optical Coherence Tomography (OCT) and with the PR of healthy subjects. Percentage of Pupil Constriction (PPC), maximal constriction velocity (MCV) and the latency of MCV (LMCV) were determined. Results: In response to red light, Best’s patients demonstrated reduced PPC and slower MCV compared with healthy subjects in nearly all test point locations. Severity of defects in PPC in responses to red light correlated with reduced thickness of photoreceptor layer as determined by OCT in the central, superior, nasal and temporal areas of the central retina (Pearson's r= -0.93, -0.91, -0.92, -0.85, respectively). By contrast, in response to blue light stimuli, the PPC and MCV of patients were lower than normal only in several central points. Surprisingly, the latency of MCV was shorter in patients compared with healthy subjects in response to red and blue stimuli. Conclusions: This study demonstrates the potential feasibility of using pupillometer-based chromatic perimetry for objectively assessing VF defects in patients with Best’s macular dystrophy. Our findings also suggest that chromatic pupilloperimetry may differentiate between PR mediated by cones or rods, and can specifically detect defects in macular cones.

Purpose: To assess whether modeling of central vision loss (CVL) due to glaucoma by optical coherence tomography (OCT) retinal nerve fiber (RNF) layer thickness (RNFLT) can be improved by including the location of the major inferior temporal retinal artery (ITA), a known correlate of individual RNF geometry. Methods: Pat- tern deviations of the two locations of the Humphrey 24-2 visual field (VF) known to be specifically vulnerable to glaucomatous CVL and OCT RNFLT on the corresponding circumpapillary sector around the optic nerve head within the radius of 1.73mm were retrospectively selected from 428 eyes of 428 patients of a large clinical glaucoma service. ITA was marked on the 1.73mm circle by a trained observer. Linear regression models were fitted with CVL as dependent variable and VF mean deviation (MD) plus either of (1) RNFLT, (2) ITA, and (3) their combination, respectively, as regressors. To assess CVL over all levels of glaucoma severity, the three models were compared to a null model containing only MD. A Baysian model comparison was performed with the Bayes Factor (BF) as measure of strength of evidence (BF<3: no evidence, 3-20: positive evidence, >20: strong evidence over null model). Results: Neither RNFLT (BF=0.9) nor ITA (BF=1.4) alone provided positive evidence over the null model, but their combination resulted in a model with strong evidence (BF=21.4). Conclusion: While the established circumpapillary RNFLT sector, based on population statistics, could not satisfactorily model CVL, the inclusion of a retinal parameter related to individual eye anatomy yielded a strong structure-function model.

Intraocular lenses (IOL) have experienced an expanding application over the last decades. Not only they can be used to cure cataract caused blindness, but they are also appointed to ease visual impairments (e.g. -18 – 10 dioptre or astigmatism).[1] These phake IOL require materials with very high refractive indices due to the limited space at the implanting position in the eye of the patient. This enables less invasive operations and such with smaller incisions.[2] Quinolinone derivates, like carbostyril, are currently known from drug design and as a main structural component of several antibiotics.[3] Although they show high refractive indices and good dispersions they have not yet been used in materials for ophthalmic applications. We synthesized and characterized novel high refractive index polymers containing quinolinones as the main refractive unit of the structure.[4] We showed that it was possible to build quinolinone polymers with high refractive indices up to 1.685 at 589 nm. Using this material it would theoretically be possible to reduce the lens thickness of an IOL to under 40 percent compared to a commercial hydrogel lens with a refractive index of 1.470. We also used the synthesized quinolinone acrylates to create hydrophobic copolymers with improved physical properties and high transmission in the visible spectral range. Besides the good lightfastness these copolymers also showed very low tendencies of glistening. In conclusion quinolinones show attractive performances for the usage as a component in acrylic copolymers. If the requirements for IOL keep rising in the coming years these monomers could be used to boost the refractive index of ophthalmic polymer compositions.

According to previous studies, the measurement of the human iris pigmentation can be exploited to detect certain eye pathological conditions in their early stage. In this paper, we propose an instrument and a method to perform hyperspectral quantitative measurements of the iris spectral reflectance. The system is based on a simple imaging setup, which includes a monochrome camera mounted on a standard ophthalmic microscope movement controller, a monochromator, and a flashing LED-based slit lamp. To assure quantitative measurements, the system is properly calibrated against a NIST reflectance standard. Iris reflectance images can be obtained in the spectral range 495-795 nm with a resolution of 25 nm. Each image consists of 1280 x 1024 pixels having a spatial resolution of 18 μm. Reflectance spectra can be calculated both from discrete areas of the iris and as the average of the whole iris surface. Preliminary results suggest that hyperspectral imaging of the iris can provide much more morphological and spectral information with respect to conventional qualitative colorimetric methods.

The unique organization of the corneal stromal collagen is responsible for mechanical strength and optical clarity of the eye. However, factors and reasons on formation of the corneal stroma is still not fully understood. Second-harmonic generation (SHG) is a nonlinear second order optical process occurring in noncentrosymmetric systems with a large hyperpolarizability. Through the combination of the second harmonic generation (SHG) microcopy and optimized Fourier-transform analysis, mature chicken corneas are investigated to probe the depth-dependent collagen organization of the corneal stroma. Our results show that the anterior stroma behaves like a fan-like distribution of successively and counterclockwisely rotated fibrous lamellae for paired corneas from the same chicken. However, the posterior stroma maintains a non-rotating pattern while increasing in depth. Surprisingly, the thickness of the anterior stroma remains almost constant throughout the temporal-nasal direction, but the posterior stroma does not behave the same. Through quantitative analysis, the natural transition of the anterior and posterior stroma is also determined. These findings enhance our understanding of the collagen-rich tissue in the chicken cornea model. Moreover, the Fourier-transformbased modality, in combination with SHG microscopy, serves as a promising tool to determine collagen alignment in embryonic development, tissue engineering and corneal diseases.

Most medical pathologies cause optical properties variations of the affected tissue. Most of current triage methods are based on imaging of the affected tissue. A scatterometer measures a distribution of optical properties that enables deduction of a myriad of parameters otherwise undetectable. We have built a system comprising of a multi wavelength scatterometer and a refractometer. The refractometer was used to account for aberrations of the eye and compensate for them in the scatterometric measurement. Results include the scatterometric characteristics of different targets that show a wavelength dependent distinct signature for each target.

Purpose: Optic disc tilt defined over 3D optic disc morphology has been shown to be associated with the location of initial glaucomatous damages. In this work, we study the impact of optic cup depth (OCD) on spatial patterns of visual field loss in glaucoma. Methods: Pairs of reliable Cirrus OCT scans around optic disc and Humphrey visual fields of glaucoma patients without visually significant cataract and age-related macular degeneration were selected. The most recent visit of a randomly selected eye of each patient was chosen. The OCD was automatically calculated on the superior-inferior cross sectional image passing through the optic disc center. The correlations between the mean pattern deviation (PD) of each sector in glaucoma hemifield test (GHT) and Garway-Heath scheme and OCD were evaluated for all severities glaucoma and mild glaucoma (mean deviation ≥ -5 dB), respectively. Results: 424 eyes of 424 patients passed the data reliability criteria with 346 mild glaucoma patients. For all severities glaucoma, there was no significant correlation between the mean sector PD and OCD. For mild glaucoma, OCD was uniquely correlated to the mean PD of the inferior pericentral sector (r=-0.18, p=0.01) in GHT, which was independent of mean deviation and retinal nerve fiber layer thickness (p<0.001 for both). Conclusion: OCD was uniquely correlated to the vision loss of the inferior pericentral sector in GHT and Garway- Health scheme for mild glaucoma. Future advancement of OCT imaging techniques may provide better clinical diagnosis for early glaucoma by focusing on 3D morphological variation of the optic disc.

In conventional fundus imaging devices, transpupillary illumination is used for illuminating the inside of the eye. In this method, the illumination light is directed into the posterior segment of the eye through the cornea and passes the pupillary area. As a result of sharing the pupillary area for the illumination beam and observation path, pupil dilation is typically necessary for wide-angle fundus examination, and the field of view is inherently limited. An alternative approach is to deliver light from the sclera. It is possible to image a wider retinal area with transcleral-illumination. However, the requirement of physical contact between the illumination probe and the sclera is a drawback of this method. We report here trans-palpebral illumination as a new method to deliver the light through the upper eyelid (palpebra). For this study, we used a 1.5 mm diameter fiber with a warm white LED light source. To illuminate the inside of the eye, the fiber illuminator was placed at the location corresponding to the pars plana region. A custom designed optical system was attached to a digital camera for retinal imaging. The optical system contained a 90 diopter ophthalmic lens and a 25 diopter relay lens. The ophthalmic lens collected light coming from the posterior of the eye and formed an aerial image between the ophthalmic and relay lenses. The aerial image was captured by the camera through the relay lens. An adequate illumination level was obtained to capture wide angle fundus images within ocular safety limits, defined by the ISO 15004-2: 2007 standard. This novel trans-palpebral illumination approach enables wide-angle fundus photography without eyeball contact and pupil dilation.

The pupil responses of 15 cognitively normal subjects (ages 60-74) were examined in response to 76 focal red and blue light stimuli using a chromatic multifocal pupillometer (CMP). Subjects with low cognitive scores as determined as by Montreal Cognitive Assessment testing, presented significantly weaker and sluggish pupil responses in peripheral and central locations of the visual field in response to red and blue light. Our findings suggests that the CMP may present a novel objective, non-invasive, low cost technique for early diagnosis of cognitive decline that may serve for Alzheimer Disease prevention and as sensitive outcome measure of therapeutic effects.

It is well established that major retinal diseases involve distortions of the retinal neural physiology and blood vascular structures. However, the details of distortions in retinal neurovascular coupling associated with major eye diseases are not well understood. In this study, a multi-modal optical coherence tomography (OCT) imaging system was developed to enable concurrent imaging of retinal neural activity and vascular hemodynamics. Flicker light stimulation was applied to mouse retinas to evoke retinal neural responses and hemodynamic changes. The OCT images were acquired continuously during the pre-stimulation, light-stimulation, and post-stimulation phases. Stimulus-evoked intrinsic optical signals (IOSs) and hemodynamic changes were observed over time in blood-free and blood regions, respectively. Rapid IOSs change occurred almost immediately after stimulation. Both positive and negative signals were observed in adjacent retinal areas. The hemodynamic changes showed time delays after stimulation. The signal magnitudes induced by light stimulation were observed in blood regions and did not show significant changes in blood-free regions. These differences may arise from different mechanisms in blood vessels and neural tissues in response to light stimulation. These characteristics agreed well with our previous observations in mouse retinas. Further development of the multimodal OCT may provide a new imaging method for studying how retinal structures and metabolic and neural functions are affected by age-related macular degeneration (AMD), glaucoma, diabetic retinopathy (DR), and other diseases, which promises novel noninvasive biomarkers for early disease detection and reliable treatment evaluations of eye diseases.

Recent work has shown that the biomechanical properties of tissues in the posterior eye have are critical for understanding the etiology and progression of ocular diseases. For instance, the primary risk for glaucoma is an elevated intraocular pressure (IOP). Weak tissues will deform under the large pressure, causing damage to vital tissues. In addition, scleral elasticity can influence the shape of the eye-globe, altering the axial length. In this work, we utilize a noncontact form of optical coherence elastography (OCE) to quantify the spatial distribution of biomechanical properties of the optic nerve, its surrounding tissues, and posterior sclera on the exterior of in situ porcine eyes in the whole eyeglobe configuration. The OCE measurements were taken at various IOPs to evaluate the biomechanical properties of the tissues as a function of IOP. The air-pulse induced dynamic response of the tissues was linked to Young’s modulus by a simple kinematic equation by quantified the damped natural frequency (DNF). The results show that the posterior sclera is not as stiff as the optic nerve and its surrounding tissues (~460 Hz and ~894 Hz at 10 mmHg IOP, respectively). Moreover, the scleral stiffness was generally unaffected by IOP (~460 Hz at 10 mmHg IOP as compared to ~516 Hz at 20 mmHg), whereas the optic nerve and its surrounding tissues stiffened as IOP was increased (~894 Hz at 10 mmHg to ~1221 Hz at 20 mmHg).

Diabetic retinopathy (DR) and age-related macular degeneration (AMD) are two of the leading causes of blindness and visual impairment in the world. Neovascularization results in severe vision loss in DR and AMD and, thus, there is an unmet need to identify mechanisms of pathogenesis and novel anti-angiogenic therapies. Zebrafish is a leading model organism for studying human disease pathogenesis, and the highly conserved drug activity between zebrafish and humans and their ability to readily absorb small molecules dissolved in water has benefited pharmaceutical discovery. Here, we use optical coherence tomography (OCT) and OCT angiography (OCT-A) to perform noninvasive, in vivo retinal imaging in a zebrafish model of vascular leakage. Zebrafish were treated with diethylaminobenzaldehyde (DEAB) to induce vascular leakage and imaged with OCT and OCT-A at six time points over two weeks: baseline one day before treatment and one, three, six, eight, and ten days post treatment. Longitudinal functional imaging showed significant vascular response immediately after DEAB treatment. Observed vascular changes included partial or complete vascular occlusion immediately after treatment and reperfusion during a two-week period. Increased vascular tortuosity several days post treatment indicated remodeling, and bifurcations and collateral vessel formation were also observed. In addition, significant treatment response variabilities were observed in the contralateral eye of the same animal. Anatomical and functional normalization was observed in most animals by ten days post treatment. These preliminary results motivate potential applications of OCT-A as a tool for studying pathogenesis and therapeutic screening in zebrafish models of retinal vascular disease.