Self-organizing cultured neural networks: Image analysis techniques for longitudinal tracking and modeling of the underlying network structure

Esta tesis estudia la evolución estructural de conjuntos de neuronas como la capacidad de auto-organización desde conjuntos de neuronas separadas hasta que forman una red (clusterizada) compleja. Esta tesis contribuye con el diseño e implementación de un algoritmo no supervisado de segmentación basado en grafos con un coste computacional muy bajo. Este algoritmo proporciona de forma automática la estructura completa de la red a partir de imágenes de cultivos neuronales tomadas con microscopios de fase con una resolución muy alta. La estructura de la red es representada mediante un objeto matemático (matriz) cuyos nodos representan a las neuronas o grupos de neuronas y los enlaces son las conexiones reconstruidas entre ellos. Este algoritmo extrae también otras medidas morfológicas importantes que caracterizan a las neuronas y a las neuritas. A diferencia de otros algoritmos hasta el momento, que necesitan de fluorescencia y técnicas inmunocitoquímicas, el algoritmo propuesto permite el estudio longitudinal de forma no invasiva posibilitando el estudio durante la formación de un cultivo. Además, esta tesis, estudia de forma sistemática un grupo de variables topológicas que garantizan la posibilidad de cuantificar e investigar la progresión de las características principales durante el proceso de auto-organización del cultivo. Nuestros resultados muestran la existencia de un estado concreto correspondiente a redes con configuracin small-world y la emergencia de propiedades a micro- y meso-escala de la estructura de la red. Finalmente, identificamos los procesos físicos principales que guían las transformaciones morfológicas de los cultivos y proponemos un modelo de crecimiento de red que reproduce el comportamiento cuantitativamente de las observaciones experimentales. ABSTRACT The thesis analyzes the morphological evolution of assemblies of living neurons, as they self-organize from collections of separated cells into elaborated, clustered, networks. In particular, it contributes with the design and implementation of a graph-based unsupervised segmentation algorithm, having an associated very low computational cost. The processing automatically retrieves the whole network structure from large scale phase-contrast images taken at high resolution throughout the entire life of a cultured neuronal network. The network structure is represented by a mathematical object (a matrix) in which nodes are identified neurons or neurons clusters, and links are the reconstructed connections between them. The algorithm is also able to extract any other relevant morphological information characterizing neurons and neurites. More importantly, and at variance with other segmentation methods that require fluorescence imaging from immunocyto- chemistry techniques, our measures are non invasive and entitle us to carry out a fully longitudinal analysis during the maturation of a single culture. In turn, a systematic statistical analysis of a group of topological observables grants us the possibility of quantifying and tracking the progression of the main networks characteristics during the self-organization process of the culture. Our results point to the existence of a particular state corresponding to a small-world network configuration, in which several relevant graphs micro- and meso-scale properties emerge. Finally, we identify the main physical processes taking place during the cultures morphological transformations, and embed them into a simplified growth model that quantitatively reproduces the overall set of experimental observations.

Self-organizing cultured neural networks: Image analysis techniques for longitudinal tracking and modeling of the underlying network structure (versión actualizada)

Esta tesis estudia la evolución estructural de conjuntos de neuronas como la capacidad de auto-organización desde conjuntos de neuronas separadas hasta que forman una red (clusterizada) compleja. Esta tesis contribuye con el diseño e implementación de un algoritmo no supervisado de segmentación basado en grafos con un coste computacional muy bajo. Este algoritmo proporciona de forma automática la estructura completa de la red a partir de imágenes de cultivos neuronales tomadas con microscopios de fase con una resolución muy alta. La estructura de la red es representada mediante un objeto matemático (matriz) cuyos nodos representan a las neuronas o grupos de neuronas y los enlaces son las conexiones reconstruidas entre ellos. Este algoritmo extrae también otras medidas morfológicas importantes que caracterizan a las neuronas y a las neuritas. A diferencia de otros algoritmos hasta el momento, que necesitan de fluorescencia y técnicas inmunocitoquímicas, el algoritmo propuesto permite el estudio longitudinal de forma no invasiva posibilitando el estudio durante la formación de un cultivo. Además, esta tesis, estudia de forma sistemática un grupo de variables topológicas que garantizan la posibilidad de cuantificar e investigar la progresión de las características principales durante el proceso de auto-organización del cultivo. Nuestros resultados muestran la existencia de un estado concreto correspondiente a redes con configuracin small-world y la emergencia de propiedades a micro- y meso-escala de la estructura de la red. Finalmente, identificamos los procesos físicos principales que guían las transformaciones morfológicas de los cultivos y proponemos un modelo de crecimiento de red que reproduce el comportamiento cuantitativamente de las observaciones experimentales. ABSTRACT The thesis analyzes the morphological evolution of assemblies of living neurons, as they self-organize from collections of separated cells into elaborated, clustered, networks. In particular, it contributes with the design and implementation of a graph-based unsupervised segmentation algorithm, having an associated very low computational cost. The processing automatically retrieves the whole network structure from large scale phase-contrast images taken at high resolution throughout the entire life of a cultured neuronal network. The network structure is represented by a mathematical object (a matrix) in which nodes are identified neurons or neurons clusters, and links are the reconstructed connections between them. The algorithm is also able to extract any other relevant morphological information characterizing neurons and neurites. More importantly, and at variance with other segmentation methods that require fluorescence imaging from immunocyto- chemistry techniques, our measures are non invasive and entitle us to carry out a fully longitudinal analysis during the maturation of a single culture. In turn, a systematic statistical analysis of a group of topological observables grants us the possibility of quantifying and tracking the progression of the main networks characteristics during the self-organization process of the culture. Our results point to the existence of a particular state corresponding to a small-world network configuration, in which several relevant graphs micro- and meso-scale properties emerge. Finally, we identify the main physical processes taking place during the cultures morphological transformations, and embed them into a simplified growth model that quantitatively reproduces the overall set of experimental observations.

Linear color correction for multiple illumination changes and non-overlapping cameras

Many image processing methods, such as techniques for people re-identification, assume photometric constancy between different images. This study addresses the correction of photometric variations based upon changes in background areas to correct foreground areas. The authors assume a multiple light source model where all light sources can have different colours and will change over time. In training mode, the authors learn per-location relations between foreground and background colour intensities. In correction mode, the authors apply a double linear correction model based on learned relations. This double linear correction includes a dynamic local illumination correction mapping as well as an inter-camera mapping. The authors evaluate their illumination correction by computing the similarity between two images based on the earth mover's distance. The authors compare the results to a representative auto-exposure algorithm found in the recent literature plus a colour correction one based on the inverse-intensity chromaticity. Especially in complex scenarios the authors’ method outperforms these state-of-the-art algorithms.

Optimizing panchromatic image change detection based on change index multiband image analysis

This work proposes an optimization of a semi-supervised Change Detection methodology based on a combination of Change Indices (CI) derived from an image multitemporal data set. For this purpose, SPOT 5 Panchromatic images with 2.5 m spatial resolution have been used, from which three Change Indices have been calculated. Two of them are usually known indices; however the third one has been derived considering the Kullbak-Leibler divergence. Then, these three indices have been combined forming a multiband image that has been used in as input for a Support Vector Machine (SVM) classifier where four different discriminant functions have been tested in order to differentiate between change and no_change categories. The performance of the suggested procedure has been assessed applying different quality measures, reaching in each case highly satisfactory values. These results have demonstrated that the simultaneous combination of basic change indices with others more sophisticated like the Kullback-Leibler distance, and the application of non-parametric discriminant functions like those employees in the SVM method, allows solving efficiently a change detection problem.

High-quality region-based foreground segmentation using a spatial grid of SVM classifiers

This paper presents a novel background modeling system that uses a spatial grid of Support Vector Machines classifiers for segmenting moving objects, which is a key step in many video-based consumer applications. The system is able to adapt to a large range of dynamic background situations since no parametric model or statistical distribution are assumed. This is achieved by using a different classifier per image region that learns the specific appearance of that scene region and its variations (illumination changes, dynamic backgrounds, etc.). The proposed system has been tested with a recent public database, outperforming other state-of-the-art algorithms.

Methodology of image analysis for study of the vertisols moisture content

The main problem to study vertical drainage from the moisture distribution, on a vertisol profile, is searching for suitable methods using these procedures. Our aim was to design a digital image processing methodology and its analysis to characterize the moisture content distribution of a vertisol profile. In this research, twelve soil pits were excavated on a ba re Mazic Pellic Vertisols ix of them in May 13/2011 and the rest in May 19 /2011 after a moderate rainfall event. Digital RGB images were taken from each vertisol pit using a Kodak? camera selecting a size of 1600x945 pixels. Each soil image was processed to homogenized brightness and then a spatial filter with several window sizes was applied to select the optimum one. The RGB image obtained were divided in each matrix color selecting the best thresholds for each one, maximum and minimum, to be applied and get a digital binary pattern. This one was analyzed by estimating two fractal scaling exponents box counting dimension D BC) and interface fractal dimension (D) In addition, three pre-fractal scaling coefficients were determinate at maximum resolution: total number of boxes intercepting the foreground pattern (A), fractal lacunarity (?1) and Shannon entropy S1). For all the images processed the spatial filter 9x9 was the optimum based on entropy, cluster and histogram criteria. Thresholds for each color were selected based on bimodal histograms.

Foreground segmentation in depth imagery using depth and spatial dynamic models for video surveillance applications

Low-cost systems that can obtain a high-quality foreground segmentation almostindependently of the existing illumination conditions for indoor environments are verydesirable, especially for security and surveillance applications. In this paper, a novelforeground segmentation algorithm that uses only a Kinect depth sensor is proposedto satisfy the aforementioned system characteristics. This is achieved by combininga mixture of Gaussians-based background subtraction algorithm with a new Bayesiannetwork that robustly predicts the foreground/background regions between consecutivetime steps. The Bayesian network explicitly exploits the intrinsic characteristics ofthe depth data by means of two dynamic models that estimate the spatial and depthevolution of the foreground/background regions. The most remarkable contribution is thedepth-based dynamic model that predicts the changes in the foreground depth distributionbetween consecutive time steps. This is a key difference with regard to visible imagery,where the color/gray distribution of the foreground is typically assumed to be constant.Experiments carried out on two different depth-based databases demonstrate that theproposed combination of algorithms is able to obtain a more accurate segmentation of theforeground/background than other state-of-the art approaches.

Image processing methods for human brain connectivity analysis from in-vivo diffusion MRI

The structural connectivity of the brain is considered to encode species-wise and subject-wise patterns that will unlock large areas of understanding of the human brain. Currently, diffusion MRI of the living brain enables to map the microstructure of tissue, allowing to track the pathways of fiber bundles connecting the cortical regions across the brain. These bundles are summarized in a network representation called connectome that is analyzed using graph theory. The extraction of the connectome from diffusion MRI requires a large processing flow including image enhancement, reconstruction, segmentation, registration, diffusion tracking, etc. Although a concerted effort has been devoted to the definition of standard pipelines for the connectome extraction, it is still crucial to define quality assessment protocols of these workflows. The definition of quality control protocols is hindered by the complexity of the pipelines under test and the absolute lack of gold-standards for diffusion MRI data. Here we characterize the impact on structural connectivity workflows of the geometrical deformation typically shown by diffusion MRI data due to the inhomogeneity of magnetic susceptibility across the imaged object. We propose an evaluation framework to compare the existing methodologies to correct for these artifacts including whole-brain realistic phantoms. Additionally, we design and implement an image segmentation and registration method to avoid performing the correction task and to enable processing in the native space of diffusion data. We release PySDCev, an evaluation framework for the quality control of connectivity pipelines, specialized in the study of susceptibility-derived distortions. In this context, we propose Diffantom, a whole-brain phantom that provides a solution to the lack of gold-standard data. The three correction methodologies under comparison performed reasonably, and it is difficult to determine which method is more advisable. We demonstrate that susceptibility-derived correction is necessary to increase the sensitivity of connectivity pipelines, at the cost of specificity. Finally, with the registration and segmentation tool called regseg we demonstrate how the problem of susceptibility-derived distortion can be overcome allowing data to be used in their original coordinates. This is crucial to increase the sensitivity of the whole pipeline without any loss in specificity.

Pulmonary Artery-Vein Segmentation in Real and Synthetic CT Images = Segmentación Arteria-Vena Pulmonares en Imágenes de CT Reales y Sintéticas

La tomografía axial computerizada (TAC) es la modalidad de imagen médica preferente para el estudio de enfermedades pulmonares y el análisis de su vasculatura. La segmentación general de vasos en pulmón ha sido abordada en profundidad a lo largo de los últimos años por la comunidad científica que trabaja en el campo de procesamiento de imagen; sin embargo, la diferenciación entre irrigaciones arterial y venosa es aún un problema abierto. De hecho, la separación automática de arterias y venas está considerado como uno de los grandes retos futuros del procesamiento de imágenes biomédicas. La segmentación arteria-vena (AV) permitiría el estudio de ambas irrigaciones por separado, lo cual tendría importantes consecuencias en diferentes escenarios médicos y múltiples enfermedades pulmonares o estados patológicos. Características como la densidad, geometría, topología y tamaño de los vasos sanguíneos podrían ser analizados en enfermedades que conllevan remodelación de la vasculatura pulmonar, haciendo incluso posible el descubrimiento de nuevos biomarcadores específicos que aún hoy en dípermanecen ocultos. Esta diferenciación entre arterias y venas también podría ayudar a la mejora y el desarrollo de métodos de procesamiento de las distintas estructuras pulmonares. Sin embargo, el estudio del efecto de las enfermedades en los árboles arterial y venoso ha sido inviable hasta ahora a pesar de su indudable utilidad. La extrema complejidad de los árboles vasculares del pulmón hace inabordable una separación manual de ambas estructuras en un tiempo realista, fomentando aún más la necesidad de diseñar herramientas automáticas o semiautomáticas para tal objetivo. Pero la ausencia de casos correctamente segmentados y etiquetados conlleva múltiples limitaciones en el desarrollo de sistemas de separación AV, en los cuales son necesarias imágenes de referencia tanto para entrenar como para validar los algoritmos. Por ello, el diseño de imágenes sintéticas de TAC pulmonar podría superar estas dificultades ofreciendo la posibilidad de acceso a una base de datos de casos pseudoreales bajo un entorno restringido y controlado donde cada parte de la imagen (incluyendo arterias y venas) está unívocamente diferenciada. En esta Tesis Doctoral abordamos ambos problemas, los cuales están fuertemente interrelacionados. Primero se describe el diseño de una estrategia para generar, automáticamente, fantomas computacionales de TAC de pulmón en humanos. Partiendo de conocimientos a priori, tanto biológicos como de características de imagen de CT, acerca de la topología y relación entre las distintas estructuras pulmonares, el sistema desarrollado es capaz de generar vías aéreas, arterias y venas pulmonares sintéticas usando métodos de crecimiento iterativo, que posteriormente se unen para formar un pulmón simulado con características realistas. Estos casos sintéticos, junto a imágenes reales de TAC sin contraste, han sido usados en el desarrollo de un método completamente automático de segmentación/separación AV. La estrategia comprende una primera extracción genérica de vasos pulmonares usando partículas espacio-escala, y una posterior clasificación AV de tales partículas mediante el uso de Graph-Cuts (GC) basados en la similitud con arteria o vena (obtenida con algoritmos de aprendizaje automático) y la inclusión de información de conectividad entre partículas. La validación de los fantomas pulmonares se ha llevado a cabo mediante inspección visual y medidas cuantitativas relacionadas con las distribuciones de intensidad, dispersión de estructuras y relación entre arterias y vías aéreas, los cuales muestran una buena correspondencia entre los pulmones reales y los generados sintéticamente. La evaluación del algoritmo de segmentación AV está basada en distintas estrategias de comprobación de la exactitud en la clasificación de vasos, las cuales revelan una adecuada diferenciación entre arterias y venas tanto en los casos reales como en los sintéticos, abriendo así un amplio abanico de posibilidades en el estudio clínico de enfermedades cardiopulmonares y en el desarrollo de metodologías y nuevos algoritmos para el análisis de imágenes pulmonares. ABSTRACT Computed tomography (CT) is the reference image modality for the study of lung diseases and pulmonary vasculature. Lung vessel segmentation has been widely explored by the biomedical image processing community, however, differentiation of arterial from venous irrigations is still an open problem. Indeed, automatic separation of arterial and venous trees has been considered during last years as one of the main future challenges in the field. Artery-Vein (AV) segmentation would be useful in different medical scenarios and multiple pulmonary diseases or pathological states, allowing the study of arterial and venous irrigations separately. Features such as density, geometry, topology and size of vessels could be analyzed in diseases that imply vasculature remodeling, making even possible the discovery of new specific biomarkers that remain hidden nowadays. Differentiation between arteries and veins could also enhance or improve methods processing pulmonary structures. Nevertheless, AV segmentation has been unfeasible until now in clinical routine despite its objective usefulness. The huge complexity of pulmonary vascular trees makes a manual segmentation of both structures unfeasible in realistic time, encouraging the design of automatic or semiautomatic tools to perform the task. However, this lack of proper labeled cases seriously limits in the development of AV segmentation systems, where reference standards are necessary in both algorithm training and validation stages. For that reason, the design of synthetic CT images of the lung could overcome these difficulties by providing a database of pseudorealistic cases in a constrained and controlled scenario where each part of the image (including arteries and veins) is differentiated unequivocally. In this Ph.D. Thesis we address both interrelated problems. First, the design of a complete framework to automatically generate computational CT phantoms of the human lung is described. Starting from biological and imagebased knowledge about the topology and relationships between structures, the system is able to generate synthetic pulmonary arteries, veins, and airways using iterative growth methods that can be merged into a final simulated lung with realistic features. These synthetic cases, together with labeled real CT datasets, have been used as reference for the development of a fully automatic pulmonary AV segmentation/separation method. The approach comprises a vessel extraction stage using scale-space particles and their posterior artery-vein classification using Graph-Cuts (GC) based on arterial/venous similarity scores obtained with a Machine Learning (ML) pre-classification step and particle connectivity information. Validation of pulmonary phantoms from visual examination and quantitative measurements of intensity distributions, dispersion of structures and relationships between pulmonary air and blood flow systems, show good correspondence between real and synthetic lungs. The evaluation of the Artery-Vein (AV) segmentation algorithm, based on different strategies to assess the accuracy of vessel particles classification, reveal accurate differentiation between arteries and vein in both real and synthetic cases that open a huge range of possibilities in the clinical study of cardiopulmonary diseases and the development of methodological approaches for the analysis of pulmonary images.

Image enhancement with MatLab algorithms

Nowadays, a lot of applications use digital images. For example in face recognition to detect and tag persons in photograph, for security control, and a lot of applications that can be found in smart cities, as speed control in roads or highways and cameras in traffic lights to detect drivers ignoring red light. Also in medicine digital images are used, such as x-ray, scanners, etc. These applications depend on the quality of the image obtained. A good camera is expensive, and the image obtained depends also on external factor as light. To make these applications work properly, image enhancement is as important as, for example, a good face detection algorithm. Image enhancement also can be used in normal photograph, for pictures done in bad light conditions, or just to improve the contrast of an image. There are some applications for smartphones that allow users apply filters or change the bright, colour or contrast on the pictures. This project compares four different techniques to use in image enhancement. After applying one of these techniques to an image, it will use better the whole available dynamic range. Some of the algorithms are designed for grey scale images and others for colour images. It is used Matlab software to develop and present the final results. These algorithms are Successive Means Quantization Transform (SMQT), Histogram Equalization, using Matlab function and own implemented function, and V transform. Finally, as conclusions, we can prove that Histogram equalization algorithm is the simplest of all, it has a wide variability of grey levels and it is not suitable for colour images. V transform algorithm is a good option for colour images. The algorithm is linear and requires low computational power. SMQT algorithm is non-linear, insensitive to gain and bias and it can extract structure of the data. RESUMEN. Hoy en día incontable número de aplicaciones usan imágenes digitales. Por ejemplo, para el control de la seguridad se usa el reconocimiento de rostros para detectar y etiquetar personas en fotografías o vídeos, para distintos usos de las ciudades inteligentes, como control de velocidad en carreteras o autopistas, cámaras en los semáforos para detectar a conductores haciendo caso omiso de un semáforo en rojo, etc. También en la medicina se utilizan imágenes digitales, como por ejemplo, rayos X, escáneres, etc. Todas estas aplicaciones dependen de la calidad de la imagen obtenida. Una buena cámara es cara, y la imagen obtenida depende también de factores externos como la luz. Para hacer que estas aplicaciones funciones correctamente, el tratamiento de imagen es tan importante como, por ejemplo, un buen algoritmo de detección de rostros. La mejora de la imagen también se puede utilizar en la fotografía no profesional o de consumo, para las fotos realizadas en malas condiciones de luz, o simplemente para mejorar el contraste de una imagen. Existen aplicaciones para teléfonos móviles que permiten a los usuarios aplicar filtros y cambiar el brillo, el color o el contraste en las imágenes. Este proyecto compara cuatro técnicas diferentes para utilizar el tratamiento de imagen. Se utiliza la herramienta de software matemático Matlab para desarrollar y presentar los resultados finales. Estos algoritmos son Successive Means Quantization Transform (SMQT), Ecualización del histograma, usando la propia función de Matlab y una nueva función que se desarrolla en este proyecto y, por último, una función de transformada V. Finalmente, como conclusión, podemos comprobar que el algoritmo de Ecualización del histograma es el más simple de todos, tiene una amplia variabilidad de niveles de gris y no es adecuado para imágenes en color. El algoritmo de transformada V es una buena opción para imágenes en color, es lineal y requiere baja potencia de cálculo. El algoritmo SMQT no es lineal, insensible a la ganancia y polarización y, gracias a él, se puede extraer la estructura de los datos.