Image Processing and Analysis for Preclinical and Clinical Applications

Radiomics is one of the most successful branches of research in the field of image processing and analysis, as it provides valuable quantitative information for the personalized medicine. It has the potential to discover features of the disease that cannot be appreciated with the naked eye in both preclinical and clinical studies. In general, all quantitative approaches based on biomedical images, such as positron emission tomography (PET), computed tomography (CT) and magnetic resonance imaging (MRI), have a positive clinical impact in the detection of biological processes and diseases as well as in predicting response to treatment. This Special Issue, “Image Processing and Analysis for Preclinical and Clinical Applications”, addresses some gaps in this field to improve the quality of research in the clinical and preclinical environment. It consists of fourteen peer-reviewed papers covering a range of topics and applications related to biomedical image processing and analysis

New algorithms for the analysis of live-cell images acquired in phase contrast microscopy

La détection et la caractérisation automatisée des cellules constituent un enjeu important dans de nombreux domaines de recherche tels que la cicatrisation, le développement de l'embryon et des cellules souches, l’immunologie, l’oncologie, l'ingénierie tissulaire et la découverte de nouveaux médicaments. Étudier le comportement cellulaire in vitro par imagerie des cellules vivantes et par le criblage à haut débit implique des milliers d'images et de vastes quantités de données. Des outils d'analyse automatisés reposant sur la vision numérique et les méthodes non-intrusives telles que la microscopie à contraste de phase (PCM) sont nécessaires. Comme les images PCM sont difficiles à analyser en raison du halo lumineux entourant les cellules et de la difficulté à distinguer les cellules individuelles, le but de ce projet était de développer des algorithmes de traitement d'image PCM dans Matlab® afin d’en tirer de l’information reliée à la morphologie cellulaire de manière automatisée. Pour développer ces algorithmes, des séries d’images de myoblastes acquises en PCM ont été générées, en faisant croître les cellules dans un milieu avec sérum bovin (SSM) ou dans un milieu sans sérum (SFM) sur plusieurs passages. La surface recouverte par les cellules a été estimée en utilisant un filtre de plage de valeurs, un seuil et une taille minimale de coupe afin d'examiner la cinétique de croissance cellulaire. Les résultats ont montré que les cellules avaient des taux de croissance similaires pour les deux milieux de culture, mais que celui-ci diminue de façon linéaire avec le nombre de passages. La méthode de transformée par ondelette continue combinée à l’analyse d'image multivariée (UWT-MIA) a été élaborée afin d’estimer la distribution de caractéristiques morphologiques des cellules (axe majeur, axe mineur, orientation et rondeur). Une analyse multivariée réalisée sur l’ensemble de la base de données (environ 1 million d’images PCM) a montré d'une manière quantitative que les myoblastes cultivés dans le milieu SFM étaient plus allongés et plus petits que ceux cultivés dans le milieu SSM. Les algorithmes développés grâce à ce projet pourraient être utilisés sur d'autres phénotypes cellulaires pour des applications de criblage à haut débit et de contrôle de cultures cellulaires.Automated cell detection and characterization is important in many research fields such as wound healing, embryo development, immune system studies, cancer research, parasite spreading, tissue engineering, stem cell research and drug research and testing. Studying in vitro cellular behavior via live-cell imaging and high-throughput screening involves thousands of images and vast amounts of data, and automated analysis tools relying on machine vision methods and non-intrusive methods such as phase contrast microscopy (PCM) are a necessity. However, there are still some challenges to overcome, since PCM images are difficult to analyze because of the bright halo surrounding the cells and blurry cell-cell boundaries when they are touching. The goal of this project was to develop image processing algorithms to analyze PCM images in an automated fashion, capable of processing large datasets of images to extract information related to cellular viability and morphology. To develop these algorithms, a large dataset of myoblasts images acquired in live-cell imaging (in PCM) was created, growing the cells in either a serum-supplemented (SSM) or a serum-free (SFM) medium over several passages. As a result, algorithms capable of computing the cell-covered surface and cellular morphological features were programmed in Matlab®. The cell-covered surface was estimated using a range filter, a threshold and a minimum cut size in order to look at the cellular growth kinetics. Results showed that the cells were growing at similar paces for both media, but their growth rate was decreasing linearly with passage number. The undecimated wavelet transform multivariate image analysis (UWT-MIA) method was developed, and was used to estimate cellular morphological features distributions (major axis, minor axis, orientation and roundness distributions) on a very large PCM image dataset using the Gabor continuous wavelet transform. Multivariate data analysis performed on the whole database (around 1 million PCM images) showed in a quantitative manner that myoblasts grown in SFM were more elongated and smaller than cells grown in SSM. The algorithms developed through this project could be used in the future on other cellular phenotypes for high-throughput screening and cell culture control applications

Quantifying groundwater-surface water interactions to improve the outcomes of human activities

Interactions between surface water and groundwater impact both the quality and quantity of water resources. This dissertation is focused on the interactions between surface water in streams and groundwater in hyporheic zones and shallow fluvial aquifers, and how human beings influence these interactions and are influenced by them. The three chapters of this dissertation span scales from cm to km and locations from Upstate New York to the Peruvian Andes, but are united by the goal to improve the scientific understanding of groundwater-surface water interactions (GWSWI) using novel techniques, in order to improve the outcomes of human activities for people and ecosystems. Heat is a useful tracer for quantifying GWSWI, but analyzing large amounts of raw thermal data has many challenges. Chapter 1 presents a computer program named VFLUX for processing raw temperature time series and calculating vertical water flux in shallow sub-surface-water systems. The workflow synthesizes several recent advancements in signal processing, and adds new techniques for calculating flux rates with large numbers of temperature records from high-resolution sensor profiles. The program includes functions for quantitatively evaluating the ideal spacing between sensor pairs, and for performing error and sensitivity analyses for the heat transport model due to thermal parameter uncertainty. The new method is demonstrated by processing two field temperature time series datasets collected using discrete temperature sensors and a high-resolution DTS profile. The analyses of field data show vertical flux rates significantly decreasing with depth at high-spatial resolution as the sensor profiles penetrate shallow, curved hyporheic flow paths, patterns which may have been obscured without the unique analytical abilities of VFLUX. Natural channel design restoration projects in streams often include the construction of cross-vanes, which are stone, dam-like structures that span the active channel, and are often thought to increase local hyporheic exchange. In Chapter 2, vertical hyporheic exchange flux (HEF) and redox-sensitive solutes were measured in the streambed around 4 cross-vanes with different morphologies. Observed patterns of HEF and redox conditions are not dominated by a single, downstream-directed hyporheic flow cell beneath cross-vanes. Instead, spatial patterns of moderate ( Melting tropical glaciers supply approximately half of dry season stream discharge in glacierized valleys of the Cordillera Blanca, Peru. The remainder of streamflow originates as groundwater stored in alpine meadows, moraines and talus slopes. A better understanding of the dynamics of alpine groundwater, including sources and contributions to streamflow and GWSWI, is important for making accurate estimates of glacial inputs to the hydrologic budget, and for our ability to make predictions about future water resources as glaciers retreat. The field study described in Chapter 3 focused on two high-elevation meadows in valleys of the Blanca. Tracer measurements of stream and spring discharge and groundwater-surface water exchange were combined with synoptic sampling of water isotopic and geochemical composition, in order to characterize and quantify contributions to streamflow from different geomorphic features. In a valley headwaters study site, groundwater supplied approximately half of stream discharge from the meadow, with most originating in a debris fan adjacent to the meadow and little from the meadow itself (6%); however, in at a mid-valley study site, where meadows are extensive, local groundwater has a large impact on stream flow and chemistry through large net discharge and fractional hydrologic turnover

SEGMENTATION AND INFORMATICS IN MULTIDIMENSIONAL FLUORESCENCE OPTICAL MICROSCOPY IMAGES

Recent advances in the field of optical microscopy have enabled scientists to observe and image complex biological processes across a wide range of spatial and temporal resolution, resulting in an exponential increase in optical microscopy data. Manual analysis of such large volumes of data is extremely time consuming and often impossible if the changes cannot be detected by the human eye. Naturally it is essential to design robust, accurate and high performance image processing and analysis tools to extract biologically significant results. Furthermore, the presentation of the results to the end-user, post analysis, is also an equally challenging issue, especially when the data (and/or the hypothesis) involves several spatial/hierarchical scales (e.g., tissues, cells, (sub)-nuclear components). This dissertation concentrates on a subset of such problems such as robust edge detection, automatic nuclear segmentation and selection in multi-dimensional tissue images, spatial analysis of gene localization within the cell nucleus, information visualization and the development of a computational framework for efficient and high-throughput processing of large datasets. Initially, we have developed 2D nuclear segmentation and selection algorithms which help in the development of an integrated approach for determining the preferential spatial localization of certain genes within the cell nuclei which is emerging as a promising technique for the diagnosis of breast cancer. Quantification requires accurate segmentation of 100 to 200 cell nuclei in each patient tissue sample in order to draw a statistically significant result. Thus, for large scale analysis involving hundreds of patients, manual processing is too time consuming and subjective. We have developed an integrated workflow that selects, following 2D automatic segmentation, a sub-population of accurately delineated nuclei for positioning of fluorescence in situ hybridization labeled genes of interest in tissue samples. Application of the method was demonstrated for discriminating normal and cancerous breast tissue sections based on the differential positioning of the HES5 gene. Automatic results agreed with manual analysis in 11 out of 14 cancers, all 4 normal cases and all 5 non-cancerous breast disease cases, thus showing the accuracy and robustness of the proposed approach. As a natural progression from the 2D analysis algorithms to 3D, we first developed a robust and accurate probabilistic edge detection method for 3D tissue samples since several down stream analysis procedures such as segmentation and tracking rely on the performance of edge detection. The method based on multiscale and multi-orientation steps surpasses several other conventional edge detectors in terms of its performance. Subsequently, given an appropriate edge measure, we developed an optimal graphcut-based 3D nuclear segmentation technique for samples where the cell nuclei are volume or surface labeled. It poses the problem as one of finding minimal closure in a directed graph and solves it efficiently using the maxflow-mincut algorithm. Both interactive and automatic versions of the algorithm are developed. The algorithm outperforms, in terms of three metrics that are commonly used to evaluate segmentation algorithms, a recently reported geodesic distance transform-based 3D nuclear segmentation method which in turns was reported to outperform several other popular tools that segment 3D nuclei in tissue samples. Finally, to apply some of the aforementioned methods to large microscopic datasets, we have developed a user friendly computing environment called MiPipeline which supports high throughput data analysis, data and process provenance, visual programming and seamlessly integrated information visualization of hierarchical biological data. The computational part of the environment is based on LONI Pipeline distributed computing server and the interactive information visualization makes use of several javascript based libraries to visualize an XML-based backbone file populated with essential meta-data and results

Proceedings of the Scientific-Practical Conference "Research and Development - 2016"

talent management; sensor arrays; automatic speech recognition; dry separation technology; oil production; oil waste; laser technolog

Remote Sensing

This dual conception of remote sensing brought us to the idea of preparing two different books; in addition to the first book which displays recent advances in remote sensing applications, this book is devoted to new techniques for data processing, sensors and platforms. We do not intend this book to cover all aspects of remote sensing techniques and platforms, since it would be an impossible task for a single volume. Instead, we have collected a number of high-quality, original and representative contributions in those areas

System Designs for Diabetic Foot Ulcer Image Assessment

For individuals with type 2 diabetes, diabetic foot ulcers represent a significant health issue and the wound care cost is quite high. Currently, clinicians and nurses mainly base their wound assessment on visual examination of wound size and the status of the wound tissue. This method is potentially inaccurate for wound assessment and requires extra clinical workload. In view of the prevalence of smartphones with high resolution digital camera, assessing wound healing by analyzing of real-time images using the significant computational power of today’s mobile devices is an attractive approach for managing foot ulcers. Alternatively, the smartphone may be used just for image capture and wireless transfer to a PC or laptop for image processing. To achieve accurate foot ulcer image assessment, we have developed and tested a novel automatic wound image analysis system which accomplishes the following conditions: 1) design of an easy-to-use image capture system which makes the image capture process comfortable for the patient and provides well-controlled image capture conditions; 2) synthesis of efficient and accurate algorithms for real-time wound boundary determination to measure the wound area size; 3) development of a quantitative method to assess the wound healing status based on a foot ulcer image sequence for a given patient and 4) design of a wound image assessment and management system that can be used both in the patient’s home and clinical environment in a tele-medicine fashion. In our work, the wound image is captured by the camera on the smartphone while the patient’s foot is held in place by an image capture box, which is specially design to aid patients in photographing ulcers occurring on the sole of their feet. The experimental results prove that our image capture system guarantees consistent illumination and a fixed distance between the foot and camera. These properties greatly reduce the complexity of the subsequent wound recognition and assessment. The most significant contribution of our work is the development of five different wound boundary determination approaches based on different computer vision algorithms. The first approach employs the level set algorithm to determine the wound boundary directly based on a manually set initial curve. The second and third approaches are the mean-shift segmentation based methods augmented by foot outline detection and analysis. These two approaches have been shown to be efficient to implement (especially on smartphones), prior-knowledge independent and able to provide reasonably accurate wound segmentation results given a set of well-tuned parameters. However, this method suffers from the lack of self-adaptivity due to the fact that it is not based on machine learning. Consequently, a two-stage Support Vector Machine (SVM) binary classifier based wound recognition approach is developed and implemented. This approach consists of three major steps 1) unsupervised super-pixel segmentation, 2) feature descriptor extraction for each super-pixel and 3) supervised classifier based wound boundary determination. The experimental results show that this approach provides promising performance (sensitivity: 73.3%, specificity: 95.6%) when dealing with foot ulcer images captured with our image capture box. In the third approach, we further relax the image capture constraints and generalize the application of our wound recognition system by applying the conditional random field (CRF) based model to solve the wound boundary determination. The key modules in this approach are the TextonBoost based potential learning at different scales and efficient CRF model inference to find the optimal labeling. Finally, the standard K-means clustering algorithm is applied to the determined wound area for color based wound tissue classification. To train the models used in the last two approaches, as well as to evaluate all three methods, we have collected about 100 wound images at the wound clinic in UMass Medical School by tracking 15 patients for a 2-year period, following an IRB approved protocol. The wound recognition results were compared with the ground truth generated by combining clinical labeling from three experienced clinicians. Specificity and sensitivity based measures indicate that the CRF based approach is the most reliable method despite its implementation complexity and computational demands. In addition, sample images of Moulage wound simulations are also used to increase the evaluation flexibility. The advantages and disadvantages of three approaches are described. Another important contribution of this work has been development of a healing score based mechanism for quantitative wound healing status assessment. The wound size and color composition measurements were converted to a score number ranging from 0-10, which indicates the healing trend based on comparisons of subsequent images to an initial foot ulcer image. By comparing the result of the healing score algorithm to the healing scores determined by experienced clinicians, we assess the clinical validity of our healing score algorithm. The level of agreement of our healing score with the three assessing clinicians was quantified by using the Kripendorff’s Alpha Coefficient (KAC). Finally, a collaborative wound image management system between the PC and smartphone was designed and successfully applied in the wound clinic for patients’ wound tracking purpose. This system is proven to be applicable in clinical environment and capable of providing interactive foot ulcer care in a telemedicine fashion

Automated Vascular Smooth Muscle Segmentation, Reconstruction, Classification and Simulation on Whole-Slide Histology

Histology of the microvasculature depicts detailed characteristics relevant to tissue perfusion. One important histologic feature is the smooth muscle component of the microvessel wall, which is responsible for controlling vessel caliber. Abnormalities can cause disease and organ failure, as seen in hypertensive retinopathy, diabetic ischemia, Alzheimer’s disease and improper cardiovascular development. However, assessments of smooth muscle cell content are conventionally performed on selected fields of view on 2D sections, which may lead to measurement bias. We have developed a software platform for automated (1) 3D vascular reconstruction, (2) detection and segmentation of muscularized microvessels, (3) classification of vascular subtypes, and (4) simulation of function through blood flow modeling. Vessels were stained for α-actin using 3,3\u27-Diaminobenzidine, assessing both normal (n=9 mice) and regenerated vasculature (n=5 at day 14, n=4 at day 28). 2D locally adaptive segmentation involved vessel detection, skeletonization, and fragment connection. 3D reconstruction was performed using our novel nucleus landmark-based registration. Arterioles and venules were categorized using supervised machine learning based on texture and morphometry. Simulation of blood flow for the normal and regenerated vasculature was performed at baseline and during demand based on the structural measures obtained from the above tools. Vessel medial area and vessel wall thickness were found to be greater in the normal vasculature as compared to the regenerated vasculature (p\u3c0.001) and a higher density of arterioles was found in the regenerated tissue (p\u3c0.05). Validation showed: a Dice coefficient of 0.88 (compared to manual) for the segmentations, a 3D reconstruction target registration error of 4 μm, and area under the receiver operator curve of 0.89 for vessel classification. We found 89% and 67% decreases in the blood flow through the network for the regenerated vasculature during increased oxygen demand as compared to the normal vasculature, respectively for 14 and 28 days post-ischemia. We developed a software platform for automated vasculature histology analysis involving 3D reconstruction, segmentation, and arteriole vs. venule classification. This advanced the knowledge of conventional histology sampling compared to whole slide analysis, the morphological and density differences in the regenerated vasculature, and the effect of the differences on blood flow and function