Damage to visual pathways can lead to irreversible blindness. Posterior visual pathways, located within a watershed area, are predisposed to hypoperfusion/reperfusion injury. In a novel rat model of bilateral common carotid artery occlusion (BCCAO), oxidative injury to the superior colliculus (SC), a major visual center within the watershed area was evaluated, in addition to its effects on retinal ganglion cells (RGCs). Nitrotyrosine, a footprint of peroxynitrite-mediated oxidative injury in the SC, and microtubule-associated protein 2, a dendrite marker in the retina, were assessed using immunofluorescence and confocal microscopy. Nitrotyrosine-immunoreactivity in the SC was increased 2 weeks after BCCAO compared to controls. Microtubule-associated protein 2-immunoreactivity in the central inner plexiform layer was reduced 3 weeks after BCCAO compared to controls. Global incomplete cerebral hypoperfusion/reperfusion induced oxidative injury in the SC and retrograde RGC dendritic changes. This suggests that cerebrovascular injury affecting the posterior visual pathways may contribute to vision loss in patients.

Inherited retinal degenerative diseases can affect various components of the retina leading to blindness. Five different mutant strains of chicken have been studied extensively as potential models for inherited retinal degeneration. The Smoky Joe (SJ) chicken is a sixth genetically blind strain of White Leghorns that shows various degrees of blindness at hatch and by 8 weeks post-hatch, have complete blindness for those that are homozygous. The objective of this study was to characterize the retinal degeneration in these birds by histology, both during embryonic and post-hatch development, and to the retinal function using electroretinograms (ERG). For both embryonic and post-hatch development, a significantly lower number of cells were found in the retina of blind birds compared to sighted (both p<0.0001). The significant contributor to cell number decrease was the loss of amacrine cells located in the inner nuclear layer. Photoreceptors were also found to potentially decrease in number, but at a later stage. ERG recordings revealed decreases in amplitudes of b-waves and oscillatory potentials in blind birds, but not in sighted. Both histology and ERG findings support the idea that the inner retinal cells are affected. The results indicate that degeneration in the Smoky Joe retina occurs mostly within the inner nuclear layer affecting amacrine cells. This hampers the functional capacity of the retina, causing blindness.

Adaptive optics (AO) systems make use of active optical elements, namely wavefront correctors, to improve the resolution of imaging systems by compensating for complex optical aberrations. Recently, magnetic fluid deformable mirrors (MFDM) were proposed as a novel type of wavefront correctors that offer cost and performance advantages over existing wavefront correctors. These mirrors are developed by coating the free surface of a magnetic fluid with a thin reflective film of nano-particles. The reflective surface of the mirrors can be deformed using a locally applied magnetic field and thus serves as a wavefront corrector. MFDMs have been found particularly suitable for ophthalmic imaging systems where they can be used to compensate for the complex aberrations in the eye that blur the images of the internal parts of the eye. However, their practical implementation in clinical devices is hampered by the lack of effective methods to control the shape of their deformable surface. The research work reported in this thesis presents solutions to the surface shape control problem in a MFDM that will make it possible for such devices to become integral components of retinal imaging AO systems. The first major contribution of this research is the development of an accurate analytical model of the dynamics of the mirror surface shape. The model is developed by analytically solving the coupled system of fluid-magnetic equations that govern the dynamics of the surface shape. The model is presented in state-space form and can be readily used in the development of surface shape control algorithms. The second major contribution of the research work is a novel, innovative design of the MFDM. The design change was prompted by the findings of the analytical work undertaken to develop the model mentioned above and is aimed at linearizing the response of the mirror surface. The proposed design also allows for mirror surface deflections that are many times higher than those provided by the conventional MFDM designs. A third contribution of this thesis involves the development of control algorithms that allowed the first ever use of a MFDM in a closed-loop adaptive optics system. A decentralized proportional-integral (PI) control algorithm developed based on the DC model of the wavefront corrector is presented to deal mostly with static or slowly time-varying aberrations. To improve the stability robustness of the closed-loop AO system, a decentralized robust proportional-integral-derivative (PID) controller is developed using the linear-matrix-inequalities (LMI) approach. To compensate for more complex dynamic aberrations, an Hinf controller is designed using the mixed-sensitivity Hinf design method. The proposed model, design and control algorithms are experimentally tested and validated.

The ability to measure the oxygen saturation, oximetry, of retinal blood both non-invasively and in-vivo has been a goal of eye research for years. Retinal oximetry can in principle be achieved from the measurement of the reflectance spectrum of the ocular fundus. Oximetry calculations are however complicated by the scattering of red blood cells, the different pathways of light through blood and the ocular tissues that light interacts with before exiting the eye. The goal of this thesis was to investigate the influence of red blood cell scattering for different light paths relevant to retinal oximetry. Results of in-vitro whole blood experiments found calculated oxygen saturation differences between blood samples measured under different retinal light paths, and these differences did not depend on the absorbance path length. We also showed that the calculated oxygen saturation value determined by a multiple linear regression Beer-Lambert absorbance model depended on the wavelength range chosen for analysis. The wavelength dependency on the calculated oxygen saturation value is due in part to the correlation that exists between the oxyhaemoglobin and deoxyhaemoglobin extinction coefficient spectra and to errors in the assumptions built into the Beer-Lambert absorbance model. A wavelength region with low correlation between the oxyhaemoglobin and deoxyhaemoglobin extinction coefficients was found that is hypothesized to be a good range to calculate oxygen saturation using a multiple linear regression approach.

Retinal pigment epithelium (RPE) is a principal site of pathogenesis in age-related macular de-generation (AMD). AMD is a main source of vision loss even blindness in the elderly and there is no effective treatment right now. Our aim is to describe the relationship between the morphology of RPE cells and the age and genotype of the eyes. We use principal component analysis (PCA) or functional principal component method (FPCA), support vector machine (SVM), and random forest (RF) methods to analyze the morphological data of RPE cells in mouse eyes to classify their age and genotype. Our analyses show that amongst all morphometric measures of RPE cells, cell shape measurements (eccentricity and solidity) are good for classification. But combination of cell shape and size (perimeter) provide best classification.

Introduction and Purpose: Vascular pathologies play an important role in the etiology and progression of number of ocular diseases. Many instruments are developed to monitor retinal hemodynamics, including the Canon Laser Blood Flowmeter (CLBF), in an attempt to better understand the pathophysiology of the disease (Chapter 2). The purpose of this thesis is to determine the impact of light scatter on retinal arteriolar hemodynamic measurement assessed by the CLBF as intraocular light scatter is an inevitable consequence of ageing and particularly cataract. Methodology: Chapter 4 – Artificial light scatter model: One eye from each of 10 healthy young subjects between the ages of 18 and 30 (23.6 ± 3.4) was randomly selected. To simulate light scatter, cells comprising a plastic collar and two plano lenses were filled with solutions of differing concentrations of polystyrene microspheres (Polysciences Inc., USA). 0.002%, 0.004%, 0.006%, 0.008% were prepared, as well as distilled water only. After a preliminary screening to confirm subject eligibility, seven arteriolar hemodynamic measurements were taken by randomly placing the cells between the CLBF objective lens and the subjects’ cornea. Chapter 5 – Ten patients scheduled for extracapsular cataract extraction using phacoemulsification and intraocular lens implantation between the ages of 61 and 84 (mean age 73 years, SD ± 8) were prospectively recruited. Two visits were required to complete the study; One prior to the surgery and one at least six weeks after the surgery to allow for full post-operative recovery. The severity of cataract was documented using the Lens Opacity Classification System (LOCS, III) at the first visit. Each subject underwent visual function assessment at both visits using logMAR Early Treatment Diabetic Retinopathy Study (ETDRS) visual acuity charts and the Brightness Acuity Tester (BAT). Retinal arteriolar hemodynamics were measured at both visits using the high intensity setting of the Canon Laser Blood Flowmeter. Results: Chapter 4: Our light scatter model resulted in an artifactual increase of retinal arteriolar diameter (p<0.0001) and thereby increased retinal blood flow (p<0.0001). The 0.006% and 0.008% microsphere concentrations produced significantly higher diameter and flow values than baseline. Centerline blood velocity, however, was not affected by light scatter. Retinal arteriolar diameter values were significantly less with the high intensity laser than with the low intensity laser (p=0.0007). Chapter 5: Group mean retinal arteriolar diameter and blood flow were reduced following extracapsular cataract extraction (Wilcoxon signed-rank test, p=0.022 and p=0.028 respectively); however, centerline blood velocity was unchanged (Wilcoxon signed-rank test, p=0.074). Conclusions: Using an artificial light scatter model (Chapter 3), we demonstrated that the densitometry assessment of vessel diameter is increasingly impacted as the magnitude of artificial light scatter increases; this effect can be partially negated by increasing laser intensity. We showed similar results in the presence of cataract (Chapter 4) by measuring the retinal arteriolar hemodynamics before and after removal of cataract. Care needs to be exercised in the interpretation of studies of retinal vessel diameter that use similar densitometry techniques as cataract is an inevitable consequence of aging.

Introduction and Purpose: Vascular pathologies play an important role in the etiology and progression of number of ocular diseases. Many instruments are developed to monitor retinal hemodynamics, including the Canon Laser Blood Flowmeter (CLBF), in an attempt to better understand the pathophysiology of the disease (Chapter 2). The purpose of this thesis is to determine the impact of light scatter on retinal arteriolar hemodynamic measurement assessed by the CLBF as intraocular light scatter is an inevitable consequence of ageing and particularly cataract. Methodology: Chapter 4 – Artificial light scatter model: One eye from each of 10 healthy young subjects between the ages of 18 and 30 (23.6 ± 3.4) was randomly selected. To simulate light scatter, cells comprising a plastic collar and two plano lenses were filled with solutions of differing concentrations of polystyrene microspheres (Polysciences Inc., USA). 0.002%, 0.004%, 0.006%, 0.008% were prepared, as well as distilled water only. After a preliminary screening to confirm subject eligibility, seven arteriolar hemodynamic measurements were taken by randomly placing the cells between the CLBF objective lens and the subjects’ cornea. Chapter 5 – Ten patients scheduled for extracapsular cataract extraction using phacoemulsification and intraocular lens implantation between the ages of 61 and 84 (mean age 73 years, SD ± 8) were prospectively recruited. Two visits were required to complete the study; One prior to the surgery and one at least six weeks after the surgery to allow for full post-operative recovery. The severity of cataract was documented using the Lens Opacity Classification System (LOCS, III) at the first visit. Each subject underwent visual function assessment at both visits using logMAR Early Treatment Diabetic Retinopathy Study (ETDRS) visual acuity charts and the Brightness Acuity Tester (BAT). Retinal arteriolar hemodynamics were measured at both visits using the high intensity setting of the Canon Laser Blood Flowmeter. Results: Chapter 4: Our light scatter model resulted in an artifactual increase of retinal arteriolar diameter (p<0.0001) and thereby increased retinal blood flow (p<0.0001). The 0.006% and 0.008% microsphere concentrations produced significantly higher diameter and flow values than baseline. Centerline blood velocity, however, was not affected by light scatter. Retinal arteriolar diameter values were significantly less with the high intensity laser than with the low intensity laser (p=0.0007). Chapter 5: Group mean retinal arteriolar diameter and blood flow were reduced following extracapsular cataract extraction (Wilcoxon signed-rank test, p=0.022 and p=0.028 respectively); however, centerline blood velocity was unchanged (Wilcoxon signed-rank test, p=0.074). Conclusions: Using an artificial light scatter model (Chapter 3), we demonstrated that the densitometry assessment of vessel diameter is increasingly impacted as the magnitude of artificial light scatter increases; this effect can be partially negated by increasing laser intensity. We showed similar results in the presence of cataract (Chapter 4) by measuring the retinal arteriolar hemodynamics before and after removal of cataract. Care needs to be exercised in the interpretation of studies of retinal vessel diameter that use similar densitometry techniques as cataract is an inevitable consequence of aging.

Retina located in the back of the eye is not only a vital part of human sight, but also contains valuable information that can be used in biometric security applications, or for the diagnosis of certain diseases. In order to analyze this information from retinal images, its features of blood vessels, microaneurysms and the optic disc require extraction and detection respectively. We propose a method to extract vessels called MF-FDOG. MF-FDOG consists of using two filters, Matched Filter (MF) and the first-order derivative of Gaussian (FDOG). The vessel map is extracted by applying a threshold to the response of MF, which is adaptively adjusted by the mean response of FDOG. This method allows us to better distinguish vessel objects from non-vessel objects. Microaneurysm (MA) detection is accomplished with two proposed algorithms, Multi-scale Correlation Filtering (MSCF) and Dictionary Learning (DL) with Sparse Representation Classifier (SRC). MSCF is hierarchical in nature, consisting of two levels: coarse level microaneurysm candidate detection and fine level true microaneurysm detection. In the first level, all possible microaneurysm candidates are found while the second level extracts features from each candidate and compares them to a discrimination table for decision (MA or non-MA). In Dictionary Learning with Sparse Representation Classifier, MA and non-MA objects are extracted from images and used to learn two dictionaries, MA and non-MA. Sparse Representation Classifier is then applied to each MA candidate object detected beforehand, using the two dictionaries to determine class membership. The detection result is further improved by adding a class discrimination term into the Dictionary Learning model. This approach is known as Centralized Dictionary Learning (CDL) with Sparse Representation Classifier. The optic disc (OD) is an important anatomical feature in retinal images, and its detection is vital for developing automated screening programs. Currently, there is no algorithm designed to automatically detect the OD in fundus images captured from Asians, which are larger and have thicker vessels compared to Caucasians. We propose such a method to complement current algorithms using two steps: OD vessel candidate detection and OD vessel candidate matching. The proposed extraction/detection approaches are tested in medical applications, specifically the case study of detecting diabetic retinopathy (DR). DR is a complication of diabetes that damages the retina and can lead to blindness. There are four stages of DR and is a leading cause of sight loss in industrialized nations. Using MF-FDOG, blood vessels were extracted from DR images, while DR images fed into MSCF and Dictionary and Centralized Dictionary Learning with Sparse Representation Classifier produced good microaneurysm detection results. Using a new database consisting of only Asian DR patients, we successfully tested our OD detection method. As part of future work we intend to improve existing methods such as enhancing low contrast microaneurysms and better scale selection. In additional, we will extract other features from the retina, develop a generalized OD detection method, apply Dictionary Learning with Sparse Representation Classifier to vessel extraction, and use the new image database to carry out more experiments in medical applications.

Thesis (Ph. D.)--University of Oklahoma. / Bibliography: leaves 150-172.

Thesis (Ph. D.)--University of Hong Kong, 2010. / Includes bibliographical references (p. 180-205). Also available in print.