Detection and Classification of Diabetic Retinopathy Pathologies in Fundus Images

Diabetic Retinopathy (DR) is a disease that affects up to 80% of diabetics around the world. It is the second greatest cause of blindness in the Western world, and one of the leading causes of blindness in the U.S. Many studies have demonstrated that early treatment can reduce the number of sight-threatening DR cases, mitigating the medical and economic impact of the disease. Accurate, early detection of eye disease is important because of its potential to reduce rates of blindness worldwide. Retinal photography for DR has been promoted for decades for its utility in both disease screening and clinical research studies. In recent years, several research centers have presented systems to detect pathology in retinal images. However, these approaches apply specialized algorithms to detect specific types of lesion in the retina. In order to detect multiple lesions, these systems generally implement multiple algorithms. Furthermore, some of these studies evaluate their algorithms on a single dataset, thus avoiding potential problems associated with the differences in fundus imaging devices, such as camera resolution. These methodologies primarily employ bottom-up approaches, in which the accurate segmentation of all the lesions in the retina is the basis for correct determination. A disadvantage of bottom-up approaches is that they rely on the accurate segmentation of all lesions in order to measure performance. On the other hand, top-down approaches do not depend on the segmentation of specific lesions. Thus, top-down methods can potentially detect abnormalities not explicitly used in their training phase. A disadvantage of these methods is that they cannot identify specific pathologies and require large datasets to build their training models. In this dissertation, I merged the advantages of the top-down and bottom-up approaches to detect DR with high accuracy. First, I developed an algorithm based on a top-down approach to detect abnormalities in the retina due to DR. By doing so, I was able to evaluate DR pathologies other than microaneurysms and exudates, which are the main focus of most current approaches. In addition, I demonstrated good generalization capacity of this algorithm by applying it to other eye diseases, such as age-related macular degeneration. Due to the fact that high accuracy is required for sight-threatening conditions, I developed two bottom-up approaches, since it has been proven that bottom-up approaches produce more accurate results than top-down approaches for particular structures. Consequently, I developed an algorithm to detect exudates in the macula. The presence of this pathology is considered to be a surrogate for clinical significant macular edema (CSME), a sight-threatening condition of DR. The analysis of the optic disc is usually not taken into account in DR screening systems. However, there is a pathology called neovascularization that is present in advanced stages of DR, making its detection of crucial clinical importance. In order to address this problem, I developed an algorithm to detect neovascularization in the optic disc. These algorithms are based on amplitude-modulation and frequency-modulation (AM-FM) representations, morphological image processing methods, and classification algorithms. The methods were tested on a diverse set of large databases and are considered to be the state-of the art in this field

UNRAVELLING DIABETIC RETINOPATHY THROUGH IMAGE PROCESSING, NEURAL NETWORKS AND FUZZY LOGIC – A REVIEW

One of the main causes of blindness is diabetic retinopathy (DR) and it may affect people of any ages. In these days, both young and old ages are affected by diabetes, and the di abetes is the main cause of DR. Hence, it is necessary to have an automated system with good accuracy and less computation time to diagnose and treat DR, and the automated system can simplify the work of ophthalmologists. The objective is to present an overview of various works recently in detecting and segmenting the various lesions of DR. Papers were categorized based on the diagnosing tools and the methods used for detecting early and advanced stage lesions. The early lesions of DR are microaneurysms, hemorrhages, exudates, and cotton wool spots and in the advanced stage, new and fragile blood vessels can be grown. Results have been evaluated in terms of sensitivity, specificity, accuracy and receiver operating characteristic curve. This paper analyzed the various steps and different algorithms used recently for the detection and classification of DR lesions. A comparison of performances has been made in terms of sensitivity, specificity, area under the curve, and accuracy. Suggestions, future workand the area to be improved were also discussed.Keywords: Diabetic retinopathy, Image processing, Morphological operations, Neural network, Fuzzy logic.Â

Computational Analysis of Fundus Images: Rule-Based and Scale-Space Models

Fundus images are one of the most important imaging examinations in modern ophthalmology because they are simple, inexpensive and, above all, noninvasive. Nowadays, the acquisition and storage of highresolution fundus images is relatively easy and fast. Therefore, fundus imaging has become a fundamental investigation in retinal lesion detection, ocular health monitoring and screening programmes. Given the large volume and clinical complexity associated with these images, their analysis and interpretation by trained clinicians becomes a timeconsuming task and is prone to human error. Therefore, there is a growing interest in developing automated approaches that are affordable and have high sensitivity and specificity. These automated approaches need to be robust if they are to be used in the general population to diagnose and track retinal diseases. To be effective, the automated systems must be able to recognize normal structures and distinguish them from pathological clinical manifestations. The main objective of the research leading to this thesis was to develop automated systems capable of recognizing and segmenting retinal anatomical structures and retinal pathological clinical manifestations associated with the most common retinal diseases. In particular, these automated algorithms were developed on the premise of robustness and efficiency to deal with the difficulties and complexity inherent in these images. Four objectives were considered in the analysis of fundus images. Segmentation of exudates, localization of the optic disc, detection of the midline of blood vessels, segmentation of the vascular network and detection of microaneurysms. In addition, we also evaluated the detection of diabetic retinopathy on fundus images using the microaneurysm detection method. An overview of the state of the art is presented to compare the performance of the developed approaches with the main methods described in the literature for each of the previously described objectives. To facilitate the comparison of methods, the state of the art has been divided into rulebased methods and machine learningbased methods. In the research reported in this paper, rulebased methods based on image processing methods were preferred over machine learningbased methods. In particular, scalespace methods proved to be effective in achieving the set goals. Two different approaches to exudate segmentation were developed. The first approach is based on scalespace curvature in combination with the local maximum of a scalespace blob detector and dynamic thresholds. The second approach is based on the analysis of the distribution function of the maximum values of the noise map in combination with morphological operators and adaptive thresholds. Both approaches perform a correct segmentation of the exudates and cope well with the uneven illumination and contrast variations in the fundus images. Optic disc localization was achieved using a new technique called cumulative sum fields, which was combined with a vascular enhancement method. The algorithm proved to be reliable and efficient, especially for pathological images. The robustness of the method was tested on 8 datasets. The detection of the midline of the blood vessels was achieved using a modified corner detector in combination with binary philtres and dynamic thresholding. Segmentation of the vascular network was achieved using a new scalespace blood vessels enhancement method. The developed methods have proven effective in detecting the midline of blood vessels and segmenting vascular networks. The microaneurysm detection method relies on a scalespace microaneurysm detection and labelling system. A new approach based on the neighbourhood of the microaneurysms was used for labelling. Microaneurysm detection enabled the assessment of diabetic retinopathy detection. The microaneurysm detection method proved to be competitive with other methods, especially with highresolution images. Diabetic retinopathy detection with the developed microaneurysm detection method showed similar performance to other methods and human experts. The results of this work show that it is possible to develop reliable and robust scalespace methods that can detect various anatomical structures and pathological features of the retina. Furthermore, the results obtained in this work show that although recent research has focused on machine learning methods, scalespace methods can achieve very competitive results and typically have greater independence from image acquisition. The methods developed in this work may also be relevant for the future definition of new descriptors and features that can significantly improve the results of automated methods.As imagens do fundo do olho são hoje um dos principais exames imagiológicos da oftalmologia moderna, pela sua simplicidade, baixo custo e acima de tudo pelo seu carácter nãoinvasivo. A aquisição e armazenamento de imagens do fundo do olho com alta resolução é também relativamente simples e rápida. Desta forma, as imagens do fundo do olho são um exame fundamental na identificação de alterações retinianas, monitorização da saúde ocular, e em programas de rastreio. Considerando o elevado volume e complexidade clínica associada a estas imagens, a análise e interpretação das mesmas por clínicos treinados tornase uma tarefa morosa e propensa a erros humanos. Assim, há um interesse crescente no desenvolvimento de abordagens automatizadas, acessíveis em custo, e com uma alta sensibilidade e especificidade. Estas devem ser robustas para serem aplicadas à população em geral no diagnóstico e seguimento de doenças retinianas. Para serem eficazes, os sistemas de análise têm que conseguir detetar e distinguir estruturas normais de sinais patológicos. O objetivo principal da investigação que levou a esta tese de doutoramento é o desenvolvimento de sistemas automáticos capazes de detetar e segmentar as estruturas anatómicas da retina, e os sinais patológicos retinianos associados às doenças retinianas mais comuns. Em particular, estes algoritmos automatizados foram desenvolvidos segundo as premissas de robustez e eficácia para lidar com as dificuldades e complexidades inerentes a estas imagens. Foram considerados quatro objetivos de análise de imagens do fundo do olho. São estes, a segmentação de exsudados, a localização do disco ótico, a deteção da linha central venosa dos vasos sanguíneos e segmentação da rede vascular, e a deteção de microaneurismas. De acrescentar que usando o método de deteção de microaneurismas, avaliouse também a capacidade de deteção da retinopatia diabética em imagens do fundo do olho. Para comparar o desempenho das metodologias desenvolvidas neste trabalho, foi realizado um levantamento do estado da arte, onde foram considerados os métodos mais relevantes descritos na literatura para cada um dos objetivos descritos anteriormente. Para facilitar a comparação entre métodos, o estado da arte foi dividido em metodologias de processamento de imagem e baseadas em aprendizagem máquina. Optouse no trabalho de investigação desenvolvido pela utilização de metodologias de análise espacial de imagem em detrimento de metodologias baseadas em aprendizagem máquina. Em particular, as metodologias baseadas no espaço de escalas mostraram ser efetivas na obtenção dos objetivos estabelecidos. Para a segmentação de exsudados foram usadas duas abordagens distintas. A primeira abordagem baseiase na curvatura em espaço de escalas em conjunto com a resposta máxima local de um detetor de manchas em espaço de escalas e limiares dinâmicos. A segunda abordagem baseiase na análise do mapa de distribuição de ruído em conjunto com operadores morfológicos e limiares adaptativos. Ambas as abordagens fazem uma segmentação dos exsudados de elevada precisão, além de lidarem eficazmente com a iluminação nãouniforme e a variação de contraste presente nas imagens do fundo do olho. A localização do disco ótico foi conseguida com uma nova técnica designada por campos de soma acumulativos, combinada com métodos de melhoramento da rede vascular. O algoritmo revela ser fiável e eficiente, particularmente em imagens patológicas. A robustez do método foi verificada pela sua avaliação em oito bases de dados. A deteção da linha central dos vasos sanguíneos foi obtida através de um detetor de cantos modificado em conjunto com filtros binários e limiares dinâmicos. A segmentação da rede vascular foi conseguida com um novo método de melhoramento de vasos sanguíneos em espaço de escalas. Os métodos desenvolvidos mostraram ser eficazes na deteção da linha central dos vasos sanguíneos e na segmentação da rede vascular. Finalmente, o método para a deteção de microaneurismas assenta num formalismo de espaço de escalas na deteção e na rotulagem dos microaneurismas. Para a rotulagem foi utilizada uma nova abordagem da vizinhança dos candidatos a microaneurismas. A deteção de microaneurismas permitiu avaliar também a deteção da retinopatia diabética. O método para a deteção de microaneurismas mostrou ser competitivo quando comparado com outros métodos, em particular em imagens de alta resolução. A deteção da retinopatia diabética exibiu um desempenho semelhante a outros métodos e a especialistas humanos. Os trabalhos descritos nesta tese mostram ser possível desenvolver uma abordagem fiável e robusta em espaço de escalas capaz de detetar diferentes estruturas anatómicas e sinais patológicos da retina. Além disso, os resultados obtidos mostram que apesar de a pesquisa mais recente concentrarse em metodologias de aprendizagem máquina, as metodologias de análise espacial apresentam resultados muito competitivos e tipicamente independentes do equipamento de aquisição das imagens. As metodologias desenvolvidas nesta tese podem ser importantes na definição de novos descritores e características, que podem melhorar significativamente o resultado de métodos automatizados

Automatic analysis of retinal images to aid in the diagnosis and grading of diabetic retinopathy

Diabetic retinopathy (DR) is the most common complication of diabetes mellitus and one of the leading causes of preventable blindness in the adult working population. Visual loss can be prevented from the early stages of DR, when the treatments are effective. Therefore, early diagnosis is paramount. However, DR may be clinically asymptomatic until the advanced stage, when vision is already affected and treatment may become difficult. For this reason, diabetic patients should undergo regular eye examinations through screening programs. Traditionally, DR screening programs are run by trained specialists through visual inspection of the retinal images. However, this manual analysis is time consuming and expensive. With the increasing incidence of diabetes and the limited number of clinicians and sanitary resources, the early detection of DR becomes non-viable. For this reason, computed-aided diagnosis (CAD) systems are required to assist specialists for a fast, reliable diagnosis, allowing to reduce the workload and the associated costs. We hypothesize that the application of novel, automatic algorithms for fundus image analysis could contribute to the early diagnosis of DR. Consequently, the main objective of the present Doctoral Thesis is to study, design and develop novel methods based on the automatic analysis of fundus images to aid in the screening, diagnosis, and treatment of DR. In order to achieve the main goal, we built a private database and used five retinal public databases: DRIMDB, DIARETDB1, DRIVE, Messidor and Kaggle. The stages of fundus image processing covered in this Thesis are: retinal image quality assessment (RIQA), the location of the optic disc (OD) and the fovea, the segmentation of RLs and EXs, and the DR severity grading. RIQA was studied with two different approaches. The first approach was based on the combination of novel, global features. Results achieved 91.46% accuracy, 92.04% sensitivity, and 87.92% specificity using the private database. We developed a second approach aimed at RIQA based on deep learning. We achieved 95.29% accuracy with the private database and 99.48% accuracy with the DRIMDB database. The location of the OD and the fovea was performed using a combination of saliency maps. The proposed methods were evaluated over the private database and the public databases DRIVE, DIARETDB1 and Messidor. For the OD, we achieved 100% accuracy for all databases except Messidor (99.50%). As for the fovea location, we also reached 100% accuracy for all databases except Messidor (99.67%). The joint segmentation of RLs and EXs was accomplished by decomposing the fundus image into layers. Results were computed per pixel and per image. Using the private database, 88.34% per-image accuracy (ACCi) was reached for the RL detection and 95.41% ACCi for EX detection. An additional method was proposed for the segmentation of RLs based on superpixels. Evaluating this method with the private database, we obtained 84.45% ACCi. Results were validated using the DIARETDB1 database. Finally, we proposed a deep learning framework for the automatic DR severity grading. The method was based on a novel attention mechanism which performs a separate attention of the dark and the bright structures of the retina. The Kaggle DR detection dataset was used for development and validation. The International Clinical DR Scale was considered, which is made up of 5 DR severity levels. Classification results for all classes achieved 83.70% accuracy and a Quadratic Weighted Kappa of 0.78. The methods proposed in this Doctoral Thesis form a complete, automatic DR screening system, contributing to aid in the early detection of DR. In this way, diabetic patients could receive better attention for their ocular health avoiding vision loss. In addition, the workload of specialists could be relieved while healthcare costs are reduced.La retinopatía diabética (RD) es la complicación más común de la diabetes mellitus y una de las principales causas de ceguera prevenible en la población activa adulta. El diagnóstico precoz es primordial para prevenir la pérdida visual. Sin embargo, la RD es clínicamente asintomática hasta etapas avanzadas, cuando la visión ya está afectada. Por eso, los pacientes diabéticos deben someterse a exámenes oftalmológicos periódicos a través de programas de cribado. Tradicionalmente, estos programas están a cargo de especialistas y se basan de la inspección visual de retinografías. Sin embargo, este análisis manual requiere mucho tiempo y es costoso. Con la creciente incidencia de la diabetes y la escasez de recursos sanitarios, la detección precoz de la RD se hace inviable. Por esta razón, se necesitan sistemas de diagnóstico asistido por ordenador (CAD) que ayuden a los especialistas a realizar un diagnóstico rápido y fiable, que permita reducir la carga de trabajo y los costes asociados. El objetivo principal de la presente Tesis Doctoral es estudiar, diseñar y desarrollar nuevos métodos basados en el análisis automático de retinografías para ayudar en el cribado, diagnóstico y tratamiento de la RD. Las etapas estudiadas fueron: la evaluación de la calidad de la imagen retiniana (RIQA), la localización del disco óptico (OD) y la fóvea, la segmentación de RL y EX y la graduación de la severidad de la RD. RIQA se estudió con dos enfoques diferentes. El primer enfoque se basó en la combinación de características globales. Los resultados lograron una precisión del 91,46% utilizando la base de datos privada. El segundo enfoque se basó en aprendizaje profundo. Logramos un 95,29% de precisión con la base de datos privada y un 99,48% con la base de datos DRIMDB. La localización del OD y la fóvea se realizó mediante una combinación de mapas de saliencia. Los métodos propuestos fueron evaluados sobre la base de datos privada y las bases de datos públicas DRIVE, DIARETDB1 y Messidor. Para el OD, logramos una precisión del 100% para todas las bases de datos excepto Messidor (99,50%). En cuanto a la ubicación de la fóvea, también alcanzamos un 100% de precisión para todas las bases de datos excepto Messidor (99,67%). La segmentación conjunta de RL y EX se logró descomponiendo la imagen del fondo de ojo en capas. Utilizando la base de datos privada, se alcanzó un 88,34% de precisión por imagen (ACCi) para la detección de RL y un 95,41% de ACCi para la detección de EX. Se propuso un método adicional para la segmentación de RL basado en superpíxeles. Evaluando este método con la base de datos privada, obtuvimos 84.45% ACCi. Los resultados se validaron utilizando la base de datos DIARETDB1. Finalmente, propusimos un método de aprendizaje profundo para la graduación automática de la gravedad de la DR. El método se basó en un mecanismo de atención. Se utilizó la base de datos Kaggle y la Escala Clínica Internacional de RD (5 niveles de severidad). Los resultados de clasificación para todas las clases alcanzaron una precisión del 83,70% y un Kappa ponderado cuadrático de 0,78. Los métodos propuestos en esta Tesis Doctoral forman un sistema completo y automático de cribado de RD, contribuyendo a ayudar en la detección precoz de la RD. De esta forma, los pacientes diabéticos podrían recibir una mejor atención para su salud ocular evitando la pérdida de visión. Además, se podría aliviar la carga de trabajo de los especialistas al mismo tiempo que se reducen los costes sanitarios.Escuela de DoctoradoDoctorado en Tecnologías de la Información y las Telecomunicacione

Automatic Segmentation of Optic Disc in Eye Fundus Images: A Survey

Optic disc detection and segmentation is one of the key elements for automatic retinal disease screening systems. The aim of this survey paper is to review, categorize and compare the optic disc detection algorithms and methodologies, giving a description of each of them, highlighting their key points and performance measures. Accordingly, this survey firstly overviews the anatomy of the eye fundus showing its main structural components along with their properties and functions. Consequently, the survey reviews the image enhancement techniques and also categorizes the image segmentation methodologies for the optic disc which include property-based methods, methods based on convergence of blood vessels, and model-based methods. The performance of segmentation algorithms is evaluated using a number of publicly available databases of retinal images via evaluation metrics which include accuracy and true positive rate (i.e. sensitivity). The survey, at the end, describes the different abnormalities occurring within the optic disc region

Automated microaneurysm detection algorithms applied to diabetic retinopathy retinal images

Diabetic retinopathy is the commonest cause of blindness in working age people. It is characterised and graded by the development of retinal microaneurysms, haemorrhages and exudates. The damage caused by diabetic retinopathy can be prevented if it is treated in its early stages. Therefore, automated early detection can limit the severity of the disease, improve the follow-up management of diabetic patients and assist ophthalmologists in investigating and treating the disease more efficiently. This review focuses on microaneurysm detection as the earliest clinically localised characteristic of diabetic retinopathy, a frequently observed complication in both Type 1 and Type 2 diabetes. Algorithms used for microaneurysm detection from retinal images are reviewed. A number of features used to extract microaneurysm are summarised. Furthermore, a comparative analysis of reported methods used to automatically detect microaneurysms is presented and discussed. The performance of methods and their complexity are also discussed

A review of feature-based retinal image analysis

Retinal imaging is a fundamental tool in ophthalmic diagnostics. The potential use of retinal imaging within screening programs, with consequent need to analyze large numbers of images with high throughput, is pushing the digital image analysis field to find new solutions for the extraction of specific information from the retinal image. The aim of this review is to explore the latest progress in image processing techniques able to recognize specific retinal image features. and potential features of disease. In particular, this review aims to describe publically available retinal image databases, highlight different performance evaluators commonly used within the field, outline current approaches in feature-based retinal image analysis, and to map related trends. This review found two key areas to be addressed for the future development of automatic retinal image analysis: fundus image quality and the affect image processing may impose on relevant clinical information within the images. Performance evaluators of the algorithms reviewed are very promising, however absolute values are difficult to interpret when validating system suitability for use within clinical practice

A Multi-Anatomical Retinal Structure Segmentation System For Automatic Eye Screening Using Morphological Adaptive Fuzzy Thresholding

Eye exam can be as efficacious as physical one in determining health concerns. Retina screening can be the very first clue to detecting a variety of hidden health issues including pre-diabetes and diabetes. Through the process of clinical diagnosis and prognosis; ophthalmologists rely heavily on the binary segmented version of retina fundus image; where the accuracy of segmented vessels, optic disc and abnormal lesions extremely affects the diagnosis accuracy which in turn affect the subsequent clinical treatment steps. This thesis proposes an automated retinal fundus image segmentation system composed of three segmentation subsystems follow same core segmentation algorithm. Despite of broad difference in features and characteristics; retinal vessels, optic disc and exudate lesions are extracted by each subsystem without the need for texture analysis or synthesis. For sake of compact diagnosis and complete clinical insight, our proposed system can detect these anatomical structures in one session with high accuracy even in pathological retina images. The proposed system uses a robust hybrid segmentation algorithm combines adaptive fuzzy thresholding and mathematical morphology. The proposed system is validated using four benchmark datasets: DRIVE and STARE (vessels), DRISHTI-GS (optic disc), and DIARETDB1 (exudates lesions). Competitive segmentation performance is achieved, outperforming a variety of up-to-date systems and demonstrating the capacity to deal with other heterogenous anatomical structures

Retinal Vascular Measurement Tools for Diagnostic Feature Extraction

The contributions of this work are in the development of new and state of the art algorithms for retinal image analysis including optic disc detection, tortuosity estimation, and cross-over abnormality detection. The retina is one of the only areas of the human body that blood vessels can be visualized noninvasively. Retinal imaging has become a standard in the ophthalmologist’s office because it is an easy and inexpensive way to monitor not just eye health, but also systemic vascular diseases. Changes to the retinal vasculature can be the early signs of diseases such as diabetic and hypertensive retinopathy, of which early detection can save vision, money, and improve overall health for the patient. When looking at the retinal vasculature, ophthalmologists generally rely on a qualitative assessment which can make comparisons over time or between different ophthalmologists difficult. Computer aided systems are now able to quantify what the ophthalmologist is qualitatively measuring in what they consider to be the most important features of the vasculature. These include, but are not limited to, tortuosity, arteriolar narrowing, cross-over abnormalities, and artery-vein (AV) ratio. The University of Padova has created a semi-automatic system for detecting and quantifying retinal vessels starting from optic disc detection, vessel segmentation, width estimation, tortuosity calculation, AV classification, and AV ratio. We propose a new method for optic disc detection that converts the retinal image into a graph and exploits vessel enhancement methods to calculate edge weights in finding the shortest path between pairs of points on the periphery of the image. The line segment with the maximum number of shortest paths is considered the optic disc location. The method was tested on three publicly available datasets: DRIVE, DIARETDB1, and Messidor consisting of 40, 89, and 1200 images and achieved an accuracy of 100, 98.88, and 99.42% respectively. The second contribution is a new algorithm for calculating abnormalities at AV crossing points. In retinal images, Gunn’s sign appears as a tapering of the vein at a crossing point, while Salus’s sign presents as an S-shaped curving. This work presents a method for the automatic quantification of these two signs once a crossover has been detected; combining segmentation, artery vein classification, and morphological feature extraction techniques to calculate vein widths and angles entering and exiting the crossover. Results on two datasets show separation between the two classes and that we can reliably detect and quantify these signs under the right conditions. The last contribution in tortuosity consists of two parts. A comparative study was performed on several of the most popular methods for tortuosity estimation on a new vessel dataset. Results show that several methods have good Cohen’s kappa agreement with both graders, while the tortuosity density metric has the highest single metric average agreement across vessel type and grader. The second is a new way to enhance curvature in segmented vessels based on a difference of Gabor filters to create a curvature enhanced image. The proposed method was tested on the RET-TORT database using several methods to calculate tortuosity, and had best Pearson’s correlation of .94 for arteries and .882 for veins, outperforming single mathematical formulations on the data. This held true after testing the method on the propose dataset as well, having higher correlation values across grader and vessel type compared with other tortuosity metrics. Summary of Results: The optic disc detection method was tested on three publicly available datasets: DRIVE, DIARETDB1, and Messidor consisting of 40, 89, and 1200 images and achieved an accuracy of 100, 98.88, and 99.42% respectively. The AV nicking quantification method was tested on a small dataset of 10 crossing provided by doctors at Papageorgiou Hospital, Aristotle University of Thessaloniki, Thessaloniki, Greece. Results showed separation between the normal and abnormal classes for both the Gunn and Salus sign. The method was then tested on a larger, publicly available dataset which showed good separation for the Gunn sign. The proposed tortuosity method was tested on the RET-TORT database using several methods to calculate tortuosity, and had best Pearson’s correlation of .94 for arteries and .882 for veins, outperforming single mathematical formulations on the data. It was then tested on the dataset proposed in this thesis, further corroborating the effectiveness of the method