Automatic quantitative analysis of morphology of apoptotic HL-60 cells

Morphological identification is a widespread procedure to assess the presence of apoptosis by visual inspection of the morphological characteristics or the fluorescence images. The procedure is lengthy and results are observer dependent. A quantitative automatic analysis is objective and would greatly help the routine work. We developed an image processing and segmentation method which combined the Otsu thresholding and morphological operators for apoptosis study. An automatic determination method of apoptotic stages of HL-60 cells with fluorescence images was developed. Comparison was made between normal cells, early apoptotic cells and late apoptotic cells about their geometric parameters which were defined to describe the features of cell morphology. The results demonstrated that the parameters we chose are very representative of the morphological characteristics of apoptotic cells. Significant differences exist between the cells in different stages, and automatic quantification of the differences can be achieved

Extraction of Nucleolus Candidate Zone in White Blood Cells of Peripheral Blood Smear Images Using Curvelet Transform

The main part of each white blood cell (WBC) is its nucleus which contains chromosomes. Although white blood cells (WBCs) with giant nuclei are the main symptom of leukemia, they are not sufficient to prove this disease and other symptoms must be investigated. For example another important symptom of leukemia is the existence of nucleolus in nucleus. The nucleus contains chromatin and a structure called the nucleolus. Chromatin is DNA in its active form while nucleolus is composed of protein and RNA, which are usually inactive. In this paper, to diagnose this symptom and in order to discriminate between nucleoli and chromatins, we employ curvelet transform, which is a multiresolution transform for detecting 2D singularities in images. For this reason, at first nuclei are extracted by means of K-means method, then curvelet transform is applied on extracted nuclei and the coefficients are modified, and finally reconstructed image is used to extract the candidate locations of chromatins and nucleoli. This method is applied on 100 microscopic images and succeeds with specificity of 80.2% and sensitivity of 84.3% to detect the nucleolus candidate zone. After nucleolus candidate zone detection, new features that can be used to classify atypical and blast cells such as gradient of saturation channel are extracted

Automated Multi-Stage Segmentation of White Blood Cells Via Optimizing Color Processing

Segmentation of white blood cells (i.e. leukocytes) is a crucial step toward the development of haematological images analysis of peripheral blood smears due to the complex nature of the different types of white blood cells and their large variations in shape, texture, color, and density. This study addresses this issue and presents a single fully automatic segmentation framework for both nuclei and cytoplasm of the five classes of leukocytes in a microscope blood smears. The proposed framework integrates a priori information of enhanced nuclei color with Gram-Schmidt orthogonalization, and multi-scale morphological enhancement to localize the nuclei, whereas clustering-based seed extraction and watershed are utilized to segment the cytoplasm. The experimental results on two different datasets show that the proposed method works successfully for both nuclei and cytoplasm segmentation, and achieves more accurate segmentation results compared to the other methods in the literature

Unsupervised segmentation technique for acute leukemia cells using clustering algorithms

Leukaemia is a blood cancer disease that contributes to the increment of mortality rate in Malaysia each year.There are two main categories for leukaemia, which are acute and chronic leukaemia.The production and development of acute leukaemia cells occurs rapidly and uncontrollable. Therefore, if the identification of acute leukaemia cells could be done fast and effectively, proper treatment and medicine could be delivered. Due to the requirement of prompt and accurate diagnosis of leukaemia, the current study has proposed unsupervised pixel segmentation based on clustering algorithm in order to obtain a fully segmented abnormal white blood cell (blast) in acute leukaemia image.In order to obtain the segmented blast, the current study proposed three clustering algorithms which are k-means, fuzzy c-means and moving k-means algorithms have been applied on the saturation component image. Then, median filter and seeded region growing area extraction algorithms have been applied, to smooth the region of segmented blast and to remove the large unwanted regions from the image, respectively.Comparisons among the three clustering algorithms are made in order to measure the performance of each clustering algorithm on segmenting the blast area. Based on the good sensitivity value that has been obtained, the results indicate that moving k-means clustering algorithm has successfully produced the fully segmented blast region in acute leukaemia image. Hence, indicating that the resultant images could be helpful to haematologists for further analysis of acute leukaemia

Hematological image analysis for acute lymphoblastic leukemia detection and classification

Microscopic analysis of peripheral blood smear is a critical step in detection of leukemia.However, this type of light microscopic assessment is time consuming, inherently subjective, and is governed by hematopathologists clinical acumen and experience. To circumvent such problems, an efficient computer aided methodology for quantitative analysis of peripheral blood samples is required to be developed. In this thesis, efforts are therefore made to devise methodologies for automated detection and subclassification of Acute Lymphoblastic Leukemia (ALL) using image processing and machine learning methods.Choice of appropriate segmentation scheme plays a vital role in the automated disease recognition process. Accordingly to segment the normal mature lymphocyte and malignant lymphoblast images into constituent morphological regions novel schemes have been proposed. In order to make the proposed schemes viable from a practical and real–time stand point, the segmentation problem is addressed in both supervised and unsupervised framework. These proposed methods are based on neural network,feature space clustering, and Markov random field modeling, where the segmentation problem is formulated as pixel classification, pixel clustering, and pixel labeling problem respectively. A comprehensive validation analysis is presented to evaluate the performance of four proposed lymphocyte image segmentation schemes against manual segmentation results provided by a panel of hematopathologists. It is observed that morphological components of normal and malignant lymphocytes differ significantly. To automatically recognize lymphoblasts and detect ALL in peripheral blood samples, an efficient methodology is proposed.Morphological, textural and color features are extracted from the segmented nucleus and cytoplasm regions of the lymphocyte images. An ensemble of classifiers represented as EOC3 comprising of three classifiers shows highest classification accuracy of 94.73% in comparison to individual members. The subclassification of ALL based on French–American–British (FAB) and World Health Organization (WHO) criteria is essential for prognosis and treatment planning. Accordingly two independent methodologies are proposed for automated classification of malignant lymphocyte (lymphoblast) images based on morphology and phenotype. These methods include lymphoblast image segmentation, nucleus and cytoplasm feature extraction, and efficient classification

Methodology for automatic classification of atypical lymphoid cells from peripheral blood cell images

Morphological analysis is the starting point for the diagnostic approach of more than 80% of the hematological diseases. However, the morphological differentiation among different types of abnormal lymphoid cells in peripheral blood is a difficult task, which requires high experience and skill. Objective values do not exist to define cytological variables, which sometimes results in doubts on the correct cell classification in the daily hospital routine. Automated systems exist which are able to get an automatic preclassification of the normal blood cells, but fail in the automatic recognition of the abnormal lymphoid cells. The general objective of this thesis is to develop a complete methodology to automatically recognize images of normal and reactive lymphocytes, and several types of neoplastic lymphoid cells circulating in peripheral blood in some mature B-cell neoplasms using digital image processing methods. This objective follows two directions: (1) with engineering and mathematical background, transversal methodologies and software tools are developed; and (2) with a view towards the clinical laboratory diagnosis, a system prototype is built and validated, whose input is a set of pathological cell images from individual patients, and whose output is the automatic classification in one of the groups of the different pathologies included in the system. This thesis is the evolution of various works, starting with a discrimination between normal lymphocytes and two types of neoplastic lymphoid cells, and ending with the design of a system for the automatic recognition of normal lymphocytes and five types of neoplastic lymphoid cells. All this work involves the development of a robust segmentation methodology using color clustering, which is able to separate three regions of interest: cell, nucleus and peripheral zone around the cell. A complete lymphoid cell description is developed by extracting features related to size, shape, texture and color. To reduce the complexity of the process, a feature selection is performed using information theory. Then, several classifiers are implemented to automatically recognize different types of lymphoid cells. The best classification results are achieved using support vector machines with radial basis function kernel. The methodology developed, which combines medical, engineering and mathematical backgrounds, is the first step to design a practical hematological diagnosis support tool in the near future.Los análisis morfológicos son el punto de partida para la orientación diagnóstica en más del 80% de las enfermedades hematológicas. Sin embargo, la clasificación morfológica entre diferentes tipos de células linfoides anormales en la sangre es una tarea difícil que requiere gran experiencia y habilidad. No existen valores objetivos para definir variables citológicas, lo que en ocasiones genera dudas en la correcta clasificación de las células en la práctica diaria en un laboratorio clínico. Existen sistemas automáticos que realizan una preclasificación automática de las células sanguíneas, pero no son capaces de diferenciar automáticamente las células linfoides anormales. El objetivo general de esta tesis es el desarrollo de una metodología completa para el reconocimiento automático de imágenes de linfocitos normales y reactivos, y de varios tipos de células linfoides neoplásicas circulantes en sangre periférica en algunos tipos de neoplasias linfoides B maduras, usando métodos de procesamiento digital de imágenes. Este objetivo sigue dos direcciones: (1) con una orientación propia de la ingeniería y la matemática de soporte, se desarrollan las metodologías transversales y las herramientas de software para su implementación; y (2) con un enfoque orientado al diagnóstico desde el laboratorio clínico, se construye y se valida un prototipo de un sistema cuya entrada es un conjunto de imágenes de células patológicas de pacientes analizados de forma individual, obtenidas mediante microscopía y cámara digital, y cuya salida es la clasificación automática en uno de los grupos de las distintas patologías incluidas en el sistema. Esta tesis es el resultado de la evolución de varios trabajos, comenzando con una discriminación entre linfocitos normales y dos tipos de células linfoides neoplásicas, y terminando con el diseño de un sistema para el reconocimiento automático de linfocitos normales y reactivos, y cinco tipos de células linfoides neoplásicas. Todo este trabajo involucra el desarrollo de una metodología de segmentación robusta usando agrupamiento por color, la cual es capaz de separar tres regiones de interés: la célula, el núcleo y la zona externa alrededor de la célula. Se desarrolla una descripción completa de la célula linfoide mediante la extracción de descriptores relacionados con el tamaño, la forma, la textura y el color. Para reducir la complejidad del proceso, se realiza una selección de descriptores usando teoría de la información. Posteriormente, se implementan varios clasificadores para reconocer automáticamente diferentes tipos de células linfoides. Los mejores resultados de clasificación se logran utilizando máquinas de soporte vectorial con núcleo de base radial. La metodología desarrollada, que combina conocimientos médicos, matemáticos y de ingeniería, es el primer paso para el diseño de una herramienta práctica de soporte al diagnóstico hematológico en un futuro cercano

Computer aided diagnosis algorithms for digital microscopy

Automatic analysis and information extraction from an image is still a highly chal- lenging research problem in the computer vision area, attempting to describe the image content with computational and mathematical techniques. Moreover the in- formation extracted from the image should be meaningful and as most discrimi- natory as possible, since it will be used to categorize its content according to the analysed problem. In the Medical Imaging domain this issue is even more felt because many important decisions that affect the patient care, depend on the use- fulness of the information extracted from the image. Manage medical image is even more complicated not only due to the importance of the problem, but also because it needs a fair amount of prior medical knowledge to be able to represent with data the visual information to which pathologist refer. Today medical decisions that impact patient care rely on the results of laboratory tests to a greater extent than ever before, due to the marked expansion in the number and complexity of offered tests. These developments promise to improve the care of patients, but the more increase the number and complexity of the tests, the more increases the possibility to misapply and misinterpret the test themselves, leading to inappropriate diagnosis and therapies. Moreover, with the increased number of tests also the amount of data to be analysed increases, forcing pathologists to devote much time to the analysis of the tests themselves rather than to patient care and the prescription of the right therapy, especially considering that most of the tests performed are just check up tests and most of the analysed samples come from healthy patients. Then, a quantitative evaluation of medical images is really essential to overcome uncertainty and subjectivity, but also to greatly reduce the amount of data and the timing for the analysis. In the last few years, many computer assisted diagno- sis systems have been developed, attempting to mimic pathologists by extracting features from the images. Image analysis involves complex algorithms to identify and characterize cells or tissues using image pattern recognition technology. This thesis addresses the main problems associated to the digital microscopy analysis in histology and haematology diagnosis, with the development of algorithms for the extraction of useful information from different digital images, but able to distinguish different biological structures in the images themselves. The proposed methods not only aim to improve the degree of accuracy of the analysis, and reducing time, if used as the only means of diagnoses, but also they can be used as intermediate tools for skimming the number of samples to be analysed directly from the pathologist, or as double check systems to verify the correct results of the automated facilities used today

Computer aided diagnosis algorithms for digital microscopy

Automatic analysis and information extraction from an image is still a highly chal- lenging research problem in the computer vision area, attempting to describe the image content with computational and mathematical techniques. Moreover the in- formation extracted from the image should be meaningful and as most discrimi- natory as possible, since it will be used to categorize its content according to the analysed problem. In the Medical Imaging domain this issue is even more felt because many important decisions that affect the patient care, depend on the use- fulness of the information extracted from the image. Manage medical image is even more complicated not only due to the importance of the problem, but also because it needs a fair amount of prior medical knowledge to be able to represent with data the visual information to which pathologist refer. Today medical decisions that impact patient care rely on the results of laboratory tests to a greater extent than ever before, due to the marked expansion in the number and complexity of offered tests. These developments promise to improve the care of patients, but the more increase the number and complexity of the tests, the more increases the possibility to misapply and misinterpret the test themselves, leading to inappropriate diagnosis and therapies. Moreover, with the increased number of tests also the amount of data to be analysed increases, forcing pathologists to devote much time to the analysis of the tests themselves rather than to patient care and the prescription of the right therapy, especially considering that most of the tests performed are just check up tests and most of the analysed samples come from healthy patients. Then, a quantitative evaluation of medical images is really essential to overcome uncertainty and subjectivity, but also to greatly reduce the amount of data and the timing for the analysis. In the last few years, many computer assisted diagno- sis systems have been developed, attempting to mimic pathologists by extracting features from the images. Image analysis involves complex algorithms to identify and characterize cells or tissues using image pattern recognition technology. This thesis addresses the main problems associated to the digital microscopy analysis in histology and haematology diagnosis, with the development of algorithms for the extraction of useful information from different digital images, but able to distinguish different biological structures in the images themselves. The proposed methods not only aim to improve the degree of accuracy of the analysis, and reducing time, if used as the only means of diagnoses, but also they can be used as intermediate tools for skimming the number of samples to be analysed directly from the pathologist, or as double check systems to verify the correct results of the automated facilities used today

Computerised diagnosis of malaria

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