Image Analysis and Processing with Applications in Proteomics and Medicine

This thesis introduces unsupervised image analysis algorithms for the segmentation of several types of images, with an emphasis on proteomics and medical images. Segmentation is a challenging task in computer vision with essential applications in biomedical engineering, remote sensing, robotics and automation. Typically, the target region is separated from the rest of image regions utilizing defining features including intensity, texture, color or motion cues. In this light, multiple segments are generated and the selection of the most significant segments becomes a controversial decision as it highly hinges on heuristic considerations. Moreover, the separation of the target regions is impeded by several daunting factors such as: background clutter, the presence of noise and artifacts as well as occlusions on multiple target regions. This thesis focuses on image segmentation using deformable models and specifically region-based Active Contours (ACs) because of their strong mathematical foundation and their appealing properties

Unsupervised level set parameterization using multi-scale filtering

This paper presents a novel framework for unsupervised level set parameterization using multi-scale filtering. A standard multi-scale, directional filtering algorithm is used in order to capture the orientation coherence in edge regions. The latter is encoded in entropy-based image `heatmaps', which are able to weight forces guiding level set evolution. Experiments are conducted on two large benchmark databases as well as on real proteomics images. The experimental results demonstrate that the proposed framework is capable of accelerating contour convergence, whereas it obtains a segmentation quality comparable to the one obtained with empirically optimized parameterization

Image Analysis and Processing With Applications in Proteomics and Medicine

Στην παρούσα διατριβή παρουσιάζονται αυτόματοι αλγόριθμοι ανάλυσης εικόνας για την κατάτμηση διαφόρων τύπων εικόνων, με έμφαση στις εικόνες πρωτεομικής και στις ιατρικές εικόνες. Οι προτεινόμενοι αλγόριθμοι βασίζονται στις αρχές των παραμορφώσιμων μοντέλων. Η διατριβή εστιάζει σε δύο κυρίως στόχους: 1) στην επίλυση του σημαντικού προβλήματος της αυτόματης παραμετροποίησης στην κατάτμηση εικόνας, 2) στην διατύπωση ενός ολοκληρωμένου μοντέλου κατάτμησης εικόνων πρωτεομικής. Η πρώτη συνεισφορά είναι ένα πρωτότυπο πλαίσιο αυτόματης παραμετροποίησης των ενεργών περιγραμμάτων περιοχής. Το πλαίσιο εμπλουτίζει τα αποτελέσματα με αντικειμενικότητα και απελευθερώνει τους τελικούς χρήστες από την επίπονη διαδικασία της εμπειρικής ρύθμισης. Εφαρμόζεται σε διάφορους τύπους ιατρικών εικόνων και παραμένει ανεπηρέαστο στις τροποποιήσεις των ρυθμίσεων των συσκευών λήψης των εικόνων αυτών. Τα πειραματικά αποτελέσματα καταδεικνύουν ότι το προτεινόμενο πλαίσιο διατηρεί υψηλή την ποιότητα κατάτμησης, συγκρίσιμη με εκείνη που επιτυγχάνεται με εμπειρική παραμετροποίηση. Η δεύτερη συνεισφορά είναι ένα αυτόματο μοντέλο βασιζόμενο στα ενεργά περιγράμματα για την κατάτμηση εικόνων πρωτεομικής. Το μοντέλο αντιμετωπίζει σημαντικά προβλήματα συμπεριλαμβανομένων των γραμμών, τεχνουργημάτων, αχνών και επικαλυπτομένων κηλίδων. Ακόμη, παρέχει εναλλακτική λύση στην επιρρεπή σε σφάλματα διαδικασία της χειρωνακτικής επεξεργασίας που απαιτείται στα υπάρχοντα πακέτα λογισμικού. Τα πειραματικά αποτελέσματα καταδεικνύουν ότι το προτεινόμενο μοντέλο υπερτερεί των υπαρχόντων πακέτων λογισμικού σε ποσοτικές μετρικές εντοπισμού και κατάτμησης.This thesis introduces unsupervised image analysis algorithms for the segmentation of several types of images, with an emphasis on proteomics and medical images. Τhe presented algorithms are tailored upon the principles of deformable models. Two objectives are pursued: 1) the core issue of unsupervised parameterization in image segmentation, 2) the formulation of a complete model for the segmentation of proteomics images. The first contribution is a novel framework for automated parameterization of region-based active contours. The presented framework endows segmentation results with objectivity and sets domain users free from the cumbersome process of empirical adjustment. It is applicable on various medical imaging modalities and remains insensitive on alterations in the settings of acquisition devices. The experimental results demonstrate that the presented framework maintains a high segmentation quality, comparable to the one obtained with empirical parameterization. The second contribution is an unsupervised active contour-based model for the segmentation of proteomics images. The presented model copes with crucial issues including streaks, artifacts, faint and overlapping spots. Moreover, it provides an alternate to the error-prone process of manual editing, required in state-of-the-art software packages. The experimental results demonstrate that the proposed model outperforms software packages in terms of detection and segmentation quantity metrics

image analysis and processing with applications in proteomics and medicine

This thesis introduces unsupervised image analysis algorithms for the segmentation of several types of images, with an emphasis on proteomics and medical images. Τhe presented algorithms are tailored upon the principles of deformable models and more specific region-based active contours. Two different objectives are pursued. The first is the core issue of unsupervised parameterization in image segmentation, whereas the second is the formulation of a complete model for the segmentation of proteomics images, which is the first to exploit the appealing attributes of active contours. The first major contribution of this thesis is a novel framework for the automated parameterization of region-based active contours. The presented framework aims to endow segmentation results with objectivity and robustness as well as to set domain users free from the cumbersome and time-consuming process of empirical adjustment. It is applicable on various medical imaging modalities and remains insensitive on alterations in the settings of the acquisition devices. The experimental results demonstrate that the presented framework maintains a segmentation quality which is comparable to the one obtained with empirical parameterization. The second major contribution of this thesis is an unsupervised active contour-based model for the segmentation of proteomics images. The presented model copes with crucial issues in 2D-GE image analysis including streaks, artifacts, faint and overlapping spots. In addition, it provides an alternate to the laborious, error-prone process of manual editing, which is required in state-of-the-art 2D-GE image analysis software packages. The experimental results demonstrate that the presented model outperforms 2D-GE image analysis software packages in terms of detection and segmentation quantity metrics

Molecular Predictors of 3D Morphogenesis by Breast Cancer Cell Lines in 3D Culture

Correlative analysis of molecular markers with phenotypic signatures is the simplest model for hypothesis generation. In this paper, a panel of 24 breast cell lines was grown in 3D culture, their morphology was imaged through phase contrast microscopy, and computational methods were developed to segment and represent each colony at multiple dimensions. Subsequently, subpopulations from these morphological responses were identified through consensus clustering to reveal three clusters of round, grape-like, and stellate phenotypes. In some cases, cell lines with particular pathobiological phenotypes clustered together (e.g., ERBB2 amplified cell lines sharing the same morphometric properties as the grape-like phenotype). Next, associations with molecular features were realized through (i) differential analysis within each morphological cluster, and (ii) regression analysis across the entire panel of cell lines. In both cases, the dominant genes that are predictive of the morphological signatures were identified. Specifically, PPARγ has been associated with the invasive stellate morphological phenotype, which corresponds to triple-negative pathobiology. PPARγ has been validated through two supporting biological assays

Opportunities and challenges for deep learning in cell dynamics research

With the growth of artificial intelligence (AI), there has been an increase in the adoption of computer vision and deep learning (DL) techniques for the evaluation of microscopy images and movies. This adoption has not only addressed hurdles in quantitative analysis of dynamic cell biological processes, but it has also started supporting advances in drug development, precision medicine and genome-phenome mapping. Here we survey existing AI-based techniques and tools, and open-source datasets, with a specific focus on the computational tasks of segmentation, classification, and tracking of cellular and subcellular structures and dynamics. We summarise long-standing challenges in microscopy video analysis from the computational perspective and review emerging research frontiers and innovative applications for deep learning-guided automation for cell dynamics research

The Role of Ecological Interactions in Polymicrobial Biofilms and their Contribution to Multiple Antibiotic Resistance

The primary objectives of this research were to demonstrate that: 1.) antibiotic resistant bacteria can promote the survival of antibiotic sensitive organisms when grown simultaneously as biofilms in antibiotics, 2.) community-level multiple antibiotic resistance of polymicrobial consortia can lead to biofilm formation despite the presence of multiple antibiotics, and 3.) biofilms may benefit plasmid retention and heterologous protein production in the absence of selective pressure. Quantitative analyses of confocal data showed that ampicillin resistant organisms supported populations of ampicillin sensitive organisms in steady state ampicillin concentrations 13 times greater than that which would inhibit sensitive cells inoculated alone. The rate of reaction of the resistance mechanism influenced the degree of protection. Spectinomycin resistant organisms did not support their sensitive counterparts, although flow cytometry indicated that GFP production by the sensitive strain was improved. When both organisms were grown in both antibiotics, larger numbers of substratum-attached pairs at 2 hours resulted in greater biofilm formation at 48 hours. For biofilms grown in both antibiotics, a benefit to spectinomycin resistant organism’s population size was detectable, but the only benefit to ampicillin resistant organisms was in terms of GFP production. Additionally, an initial attachment ratio of 5 spectinomycin resistant organisms to 1 ampicillin resistant organism resulted in optimal biofilm formation at 48 hours. Biofilms also enhanced the stability of high-copy number plasmids and heterologous protein production. In the absence of antibiotic selective pressure, plasmid DNA was not detected after 48 hours in chemostats, where the faster growth rate of plasmid-free cells contributed to the washout of plasmid retaining cells. The plasmid copy number per cell in biofilms grown without antibiotic selective pressure steadily increased over a six day period. Flow cytometric monitoring of bacteria grown in biofilms indicated that 95 percent of the population was producing GFP at 48 hours. This research supports the idea that ecological interactions between bacteria contribute to biofilm development in the presence of antibiotics, and demonstrates that community-level multiple antibiotic resistance is a factor in biofilm recalcitrance against antibiotics. Additionally, biofilms may provide an additional tool for stabilizing high copy number plasmids used for heterologous protein production

Characterising pattern asymmetry in pigmented skin lesions

Abstract. In clinical diagnosis of pigmented skin lesions asymmetric pigmentation is often indicative of melanoma. This paper describes a method and measures for characterizing lesion symmetry. The estimate of mirror symmetry is computed first for a number of axes at different degrees of rotation with respect to the lesion centre. The statistics of these estimates are the used to assess the overall symmetry. The method is applied to three different lesion representations showing the overall pigmentation, the pigmentation pattern, and the pattern of dermal melanin. The best measure is a 100% sensitive and 96% specific indicator of melanoma on a test set of 33 lesions, with a separate training set consisting of 66 lesions

Machine learning applications in proteomics research: How the past can boost the future

Machine learning is a subdiscipline within artificial intelligence that focuses on algorithms that allow computers to learn solving a (complex) problem from existing data. This ability can be used to generate a solution to a particularly intractable problem, given that enough data are available to train and subsequently evaluate an algorithm on. Since MS-based proteomics has no shortage of complex problems, and since publicly available data are becoming available in ever growing amounts, machine learning is fast becoming a very popular tool in the field. We here therefore present an overview of the different applications of machine learning in proteomics that together cover nearly the entire wet- and dry-lab workflow, and that address key bottlenecks in experiment planning and design, as well as in data processing and analysis.acceptedVersio

Improving cancer subtype diagnosis and grading using clinical decision support system based on computer-aided tissue image analysis

This research focuses towards the development of a clinical decision support system (CDSS) based on cellular and tissue image analysis and classification system that improves consistency and facilitates the clinical decision making process. In a typical cancer examination, pathologists make diagnosis by manually reading morphological features in patient biopsy images, in which cancer biomarkers are highlighted by using different staining techniques. This process is subjected to pathologist's training and experience, especially when the same cancer has several subtypes (i.e. benign tumor subtype vs. malignant subtype) and the same cancer tissue biopsy contains heterogeneous morphologies in different locations. The variability in pathologist's manual reading may result in varying cancer diagnosis and treatment. This Ph.D. research aims to reduce the subjectivity and variation existing in traditional histo-pathological reading of patient tissue biopsy slides through Computer-Aided Diagnosis (CAD). Using the CAD, quantitative molecular profiling of cancer biomarkers of stained biopsy images are obtained by extracting and analyzing texture and cellular structure features. In addition, cancer sub-type classification and a semi-automatic grade scoring (i.e. clinical decision making) for improved consistency over a large number of cancer subtype images can be performed. The CAD tools do have their own limitations and in certain cases the clinicians, however, prefer systems which are flexible and take into account their individuality when necessary by providing some control rather than fully automated system. Therefore, to be able to introduce CDSS in health care, we need to understand users' perspectives and preferences on the new information technology. This forms as the basis for this research where we target to present the quantitative information acquired through the image analysis, annotate the images and provide suitable visualization which can facilitate the process of decision making in a clinical setting.PhDCommittee Chair: Dr. May D. Wang; Committee Member: Dr. Andrew N. Young; Committee Member: Dr. Anthony J. Yezzi; Committee Member: Dr. Edward J. Coyle; Committee Member: Dr. Paul Benkese