Haptics in forensics: the possibilities and advantages in using the haptic device for reconstruction approaches in forensic science

Non-invasive documentation methods such as surface scanning and radiological imaging are gaining in importance in the forensic field. These three-dimensional technologies provide digital 3D data, which are processed and handled in the computer. However, the sense of touch gets lost using the virtual approach. The haptic device enables the use of the sense of touch to handle and feel digital 3D data. The multifunctional application of a haptic device for forensic approaches is evaluated and illustrated in three different cases: the representation of bone fractures of the lower extremities, by traffic accidents, in a non-invasive manner; the comparison of bone injuries with the presumed injury-inflicting instrument; and in a gunshot case, the identification of the gun by the muzzle imprint, and the reconstruction of the holding position of the gun. The 3D models of the bones are generated from the Computed Tomography (CT) images. The 3D models of the exterior injuries, the injury-inflicting tools and the bone injuries, where a higher resolution is necessary, are created by the optical surface scan. The haptic device is used in combination with the software FreeForm Modelling Plus for touching the surface of the 3D models to feel the minute injuries and the surface of tools, to reposition displaced bone parts and to compare an injury-causing instrument with an injury. The repositioning of 3D models in a reconstruction is easier, faster and more precisely executed by means of using the sense of touch and with the user-friendly movement in the 3D space. For representation purposes, the fracture lines of bones are coloured. This work demonstrates that the haptic device is a suitable and efficient application in forensic science. The haptic device offers a new way in the handling of digital data in the virtual 3D space.

Optical hysteresis of ridge waveguide magnetic garnet films and cavities in two-dimensional magneto-photonic crystal slabs

Magnetic iron garnets as well as magnetic photonic crystals are of great interests in magneto-optic applications such as isolators, current captors, circulators, TE-TM mode conversion, wavelength accordable filters, optical sensors and switches, all of which provide a promising platform for future integrated optical circuits. In the present work, two topics are studied based on magnetic iron garnet films. In the first part, the characteristics of the magnetization are investigated for ridge waveguides fabricated on (100) oriented iron garnet thin films. The magnetic response in magneto-optic waveguides patterned on epitaxial magnetic garnet films depends on the crystallographic orientation of the waveguides and the magnetic anisotropy of the material. These can be studied by polarization rotation hysteresis loops, which are related to the component of magnetization parallel to the light propagation direction and the linear birefringence. Polarization rotation hysteresis loops for low birefringence waveguides with different orientations are experimentally investigated. Asymmetric stepped curves are obtained from waveguides along, due to the large magnetocrystalline anisotropy in the plane. A model based on the free energy density is developed to demonstrate the motion of the magnetization and can be used in the design of magneto-optic devices. The second part of this thesis focuses on the design and fabrication of high-Q cavities in two-dimensional magneto-photonic crystal slabs. The device consists of a layer of silicon and a layer of iron garnet thin film. Triangular lattice elliptical air holes are patterned in the slab. The fundamental TM band gap overlaps with the first-order TE band gap from 0374~0.431(a/λ) showing that both TE and TM polarization light can be confined in the photonic crystals. A nanocavity is designed to obtain both TE and TM defect modes in the band gaps. Additional work is needed to overlap the TE and TM defect modes and obtain a high-Q cavity so as to develop miniaturized Faraday rotators.

Application of MFBD Algorithms to Image Reconstruction Under Anisoplanatic Conditions

All optical systems that operate in or through the atmosphere suffer from turbulence induced image blur. Both military and civilian surveillance, gun-sighting, and target identification systems are interested in terrestrial imaging over very long horizontal paths, but atmospheric turbulence can blur the resulting images beyond usefulness. My dissertation explores the performance of a multi-frame-blind-deconvolution technique applied under anisoplanatic conditions for both Gaussian and Poisson noise model assumptions. The technique is evaluated for use in reconstructing images of scenes corrupted by turbulence in long horizontal-path imaging scenarios and compared to other speckle imaging techniques. Performance is evaluated via the reconstruction of a common object from three sets of simulated turbulence degraded imagery representing low, moderate and severe turbulence conditions. Each set consisted of 1000 simulated, turbulence degraded images. The MSE performance of the estimator is evaluated as a function of the number of images, and the number of Zernike polynomial terms used to characterize the point spread function. I will compare the mean-square-error (MSE) performance of speckle imaging methods and a maximum-likelihood, multi-frame blind deconvolution (MFBD) method applied to long-path horizontal imaging scenarios. Both methods are used to reconstruct a scene from simulated imagery featuring anisoplanatic turbulence induced aberrations. This comparison is performed over three sets of 1000 simulated images each for low, moderate and severe turbulence-induced image degradation. The comparison shows that speckle-imaging techniques reduce the MSE 46 percent, 42 percent and 47 percent on average for low, moderate, and severe cases, respectively using 15 input frames under daytime conditions and moderate frame rates. Similarly, the MFBD method provides, 40 percent, 29 percent, and 36 percent improvements in MSE on average under the same conditions. The comparison is repeated under low light conditions (less than 100 photons per pixel) where improvements of 39 percent, 29 percent and 27 percent are available using speckle imaging methods and 25 input frames and 38 percent, 34 percent and 33 percent respectively for the MFBD method and 150 input frames. The MFBD estimator is applied to three sets of field data and the results presented. Finally, a combined Bispectrum-MFBD Hybrid estimator is proposed and investigated. This technique consistently provides a lower MSE and smaller variance in the estimate under all three simulated turbulence conditions.

Automatic Data Normalization and Parameterization for Optical Motion Tracking

Methods for optical motion capture often require timeconsuming manual processing before the data can be used for subsequent tasks such as retargeting or character animation. These processing steps restrict the applicability of motion capturing especially for dynamic VR-environments with real time requirements. To solve these problems, we present two additional, fast and automatic processing stages based on our motion capture pipeline presented in [HSK05]. A normalization step aligns the recorded coordinate systems with the skeleton structure to yield a common and intuitive data basis across different recording sessions. A second step computes a parameterization based on automatically extracted main movement axes to generate a compact motion description. Our method does not restrict the placement of marker bodies nor the recording setup, and only requires a short calibration phase.

RANGE ADAPTIVE PROTON THERAPY FOR PROSTATE CANCER

Purpose: The rapid distal falloff of a proton beam allows for sparing of normal tissues distal to the target. However proton beams that aim directly towards critical structures are avoided due to concerns of range uncertainties, such as CT number conversion and anatomy variations. We propose to eliminate range uncertainty and enable prostate treatment with a single anterior beam by detecting the proton’s range at the prostate-rectal interface and adaptively adjusting the range in vivo and in real-time. Materials and Methods: A prototype device, consisting of an endorectal liquid scintillation detector and dual-inverted Lucite wedges for range compensation, was designed to test the feasibility and accuracy of the technique. Liquid scintillation filled volume was fitted with optical fiber and placed inside the rectum of an anthropomorphic pelvic phantom. Photodiode-generated current signal was generated as a function of proton beam distal depth, and the spatial resolution of this technique was calculated by relating the variance in detecting proton spills to its maximum penetration depth. The relative water-equivalent thickness of the wedges was measured in a water phantom and prospectively tested to determine the accuracy of range corrections. Treatment simulation studies were performed to test the potential dosimetric benefit in sparing the rectum. Results: The spatial resolution of the detector in phantom measurement was 0.5 mm. The precision of the range correction was 0.04 mm. The residual margin to ensure CTV coverage was 1.1 mm. The composite distal margin for 95% treatment confidence was 2.4 mm. Planning studies based on a previously estimated 2mm margin (90% treatment confidence) for 27 patients showed a rectal sparing up to 51% at 70 Gy and 57% at 40 Gy relative to IMRT and bilateral proton treatment. Conclusion: We demonstrated the feasibility of our design. Use of this technique allows for proton treatment using a single anterior beam, significantly reducing the rectal dose.

Perfusion in rat brain at 7 T with arterial spin labeling using FAIR-TrueFISP and QUIPSS.

Measurement of perfusion in longitudinal studies allows for the assessment of tissue integrity and the detection of subtle pathologies. In this work, the feasibility of measuring brain perfusion in rats with high spatial resolution using arterial spin labeling is reported. A flow-sensitive alternating recovery sequence, coupled with a balanced gradient fast imaging with steady-state precession readout section was used to minimize ghosting and geometric distortions, while achieving high signal-to-noise ratio. The quantitative imaging of perfusion using a single subtraction method was implemented to address the effects of variable transit delays between the labeling of spins and their arrival at the imaging slice. Studies in six rats at 7 T showed good perfusion contrast with minimal geometric distortion. The measured blood flow values of 152.5+/-6.3 ml/100 g per minute in gray matter and 72.3+/-14.0 ml/100 g per minute in white matter are in good agreement with previously reported values based on autoradiography, considered to be the gold standard.

Streak detection algorithm for space debris detection on optical images.

Any image processing object detection algorithm somehow tries to integrate the object light (Recognition Step) and applies statistical criteria to distinguish objects of interest from other objects or from pure background (Decision Step). There are various possibilities how these two basic steps can be realized, as can be seen in the different proposed detection methods in the literature. An ideal detection algorithm should provide high recognition sensitiv ity with high decision accuracy and require a reasonable computation effort . In reality, a gain in sensitivity is usually only possible with a loss in decision accuracy and with a higher computational effort. So, automatic detection of faint streaks is still a challenge. This paper presents a detection algorithm using spatial filters simulating the geometrical form of possible streaks on a CCD image. This is realized by image convolution. The goal of this method is to generate a more or less perfect match between a streak and a filter by varying the length and orientation of the filters. The convolution answers are accepted or rejected according to an overall threshold given by the ackground statistics. This approach yields as a first result a huge amount of accepted answers due to filters partially covering streaks or remaining stars. To avoid this, a set of additional acceptance criteria has been included in the detection method. All criteria parameters are justified by background and streak statistics and they affect the detection sensitivity only marginally. Tests on images containing simulated streaks and on real images containing satellite streaks show a very promising sensitivity, reliability and running speed for this detection method. Since all method parameters are based on statistics, the true alarm, as well as the false alarm probability, are well controllable. Moreover, the proposed method does not pose any extraordinary demands on the computer hardware and on the image acquisition process.

Detection and Differentiation of Intraretinal Hemorrhage in Spectral Domain Optical Coherence Tomography.

PURPOSE The purpose of this study was to classify and detect intraretinal hemorrhage (IRH) in spectral domain optical coherence tomography (SD-OCT). METHODS Initially the presentation of IRH in BRVO-patients in SD-OCT was described by one reader comparing color-fundus (CF) and SD-OCT using dedicated software. Based on these established characteristics, the presence and the severity of IRH in SD-OCT and CF were assessed by two other masked readers and the inter-device and the inter-observer agreement were evaluated. Further the area of IRH was compared. RESULTS About 895 single B-scans of 24 eyes were analyzed. About 61% of SD-OCT scans and 46% of the CF-images were graded for the presence of IRH (concordance: 73%, inter-device agreement: k = 0.5). However, subdivided into previously established severity levels of dense (CF: 21.3% versus SD-OCT: 34.7%, k = 0.2), flame-like (CF: 15.5% versus SD-OCT: 45.5%, k = 0.3), and dot-like (CF: 32% versus SD-OCT: 24.4%, k = 0.2) IRH, the inter-device agreement was weak. The inter-observer agreement was strong with k = 0.9 for SD-OCT and k = 0.8 for CF. The mean area of IRH detected on SD-OCT was significantly greater than on CF (SD-OCT: 11.5 ± 4.3 mm(2) versus CF: 8.1 ± 5.5 mm(2), p = 0.008). CONCLUSIONS IRH seems to be detectable on SD-OCT; however, the previously established severity grading agreed weakly with that assessed by CF.

Bio-inspired FPGA Architecture for Self-Calibration of an Image Compression Core based on Wavelet Transforms in Embedded Systems

A generic bio-inspired adaptive architecture for image compression suitable to be implemented in embedded systems is presented. The architecture allows the system to be tuned during its calibration phase. An evolutionary algorithm is responsible of making the system evolve towards the required performance. A prototype has been implemented in a Xilinx Virtex-5 FPGA featuring an adaptive wavelet transform core directed at improving image compression for specific types of images. An Evolution Strategy has been chosen as the search algorithm and its typical genetic operators adapted to allow for a hardware friendly implementation. HW/SW partitioning issues are also considered after a high level description of the algorithm is profiled which validates the proposed resource allocation in the device fabric. To check the robustness of the system and its adaptation capabilities, different types of images have been selected as validation patterns. A direct application of such a system is its deployment in an unknown environment during design time, letting the calibration phase adjust the system parameters so that it performs efcient image compression. Also, this prototype implementation may serve as an accelerator for the automatic design of evolved transform coefficients which are later on synthesized and implemented in a non-adaptive system in the final implementation device, whether it is a HW or SW based computing device. The architecture has been built in a modular way so that it can be easily extended to adapt other types of image processing cores. Details on this pluggable component point of view are also given in the paper.

Image sub-segmentation by PFCM and Artificial Neural Networks to detect pore space in 2D and 3D CT soil images

The image by Computed Tomography is a non-invasive alternative for observing soil structures, mainly pore space. The pore space correspond in soil data to empty or free space in the sense that no material is present there but only fluids, the fluid transport depend of pore spaces in soil, for this reason is important identify the regions that correspond to pore zones. In this paper we present a methodology in order to detect pore space and solid soil based on the synergy of the image processing, pattern recognition and artificial intelligence. The mathematical morphology is an image processing technique used for the purpose of image enhancement. In order to find pixels groups with a similar gray level intensity, or more or less homogeneous groups, a novel image sub-segmentation based on a Possibilistic Fuzzy c-Means (PFCM) clustering algorithm was used. The Artificial Neural Networks (ANNs) are very efficient for demanding large scale and generic pattern recognition applications for this reason finally a classifier based on artificial neural network is applied in order to classify soil images in two classes, pore space and solid soil respectively.

An approach to hand biometrics in mobile devices

This paper focuses on hand biometrics applied to images acquired from a mobile device. The system offers the possibility of identifying individuals based on features extracted from hand pictures obtained with a low-quality camera embedded on a mobile device. Furthermore, the acquisitions have been carried out regardless illumination control, orientation, distance to camera, and similar aspects. In addition, the whole system has been tested with an owned database. Finally, the results obtained (6.0% ± 0.2) and the algorithm structure are both promising in relation to a posterior mobile implementation

Distributed collaborative processing in wireless sensor networks with application to target localization and beamforming

Abstract The proliferation of wireless sensor networks and the variety of envisioned applications associated with them has motivated the development of distributed algorithms for collaborative processing over networked systems. One of the applications that has attracted the attention of the researchers is that of target localization where the nodes of the network try to estimate the position of an unknown target that lies within its coverage area. Particularly challenging is the problem of estimating the target’s position when we use received signal strength indicator (RSSI) due to the nonlinear relationship between the measured signal and the true position of the target. Many of the existing approaches suffer either from high computational complexity (e.g., particle filters) or lack of accuracy. Further, many of the proposed solutions are centralized which make their application to a sensor network questionable. Depending on the application at hand and, from a practical perspective it could be convenient to find a balance between localization accuracy and complexity. Into this direction we approach the maximum likelihood location estimation problem by solving a suboptimal (and more tractable) problem. One of the main advantages of the proposed scheme is that it allows for a decentralized implementation using distributed processing tools (e.g., consensus and convex optimization) and therefore, it is very suitable to be implemented in real sensor networks. If further accuracy is needed an additional refinement step could be performed around the found solution. Under the assumption of independent noise among the nodes such local search can be done in a fully distributed way using a distributed version of the Gauss-Newton method based on consensus. Regardless of the underlying application or function of the sensor network it is al¬ways necessary to have a mechanism for data reporting. While some approaches use a special kind of nodes (called sink nodes) for data harvesting and forwarding to the outside world, there are however some scenarios where such an approach is impractical or even impossible to deploy. Further, such sink nodes become a bottleneck in terms of traffic flow and power consumption. To overcome these issues instead of using sink nodes for data reporting one could use collaborative beamforming techniques to forward directly the generated data to a base station or gateway to the outside world. In a dis-tributed environment like a sensor network nodes cooperate in order to form a virtual antenna array that can exploit the benefits of multi-antenna communications. In col-laborative beamforming nodes synchronize their phases in order to add constructively at the receiver. Some of the inconveniences associated with collaborative beamforming techniques is that there is no control over the radiation pattern since it is treated as a random quantity. This may cause interference to other coexisting systems and fast bat-tery depletion at the nodes. Since energy-efficiency is a major design issue we consider the development of a distributed collaborative beamforming scheme that maximizes the network lifetime while meeting some quality of service (QoS) requirement at the re¬ceiver side. Using local information about battery status and channel conditions we find distributed algorithms that converge to the optimal centralized beamformer. While in the first part we consider only battery depletion due to communications beamforming, we extend the model to account for more realistic scenarios by the introduction of an additional random energy consumption. It is shown how the new problem generalizes the original one and under which conditions it is easily solvable. By formulating the problem under the energy-efficiency perspective the network’s lifetime is significantly improved. Resumen La proliferación de las redes inalámbricas de sensores junto con la gran variedad de posi¬bles aplicaciones relacionadas, han motivado el desarrollo de herramientas y algoritmos necesarios para el procesado cooperativo en sistemas distribuidos. Una de las aplicaciones que suscitado mayor interés entre la comunidad científica es la de localization, donde el conjunto de nodos de la red intenta estimar la posición de un blanco localizado dentro de su área de cobertura. El problema de la localization es especialmente desafiante cuando se usan niveles de energía de la seal recibida (RSSI por sus siglas en inglés) como medida para la localization. El principal inconveniente reside en el hecho que el nivel de señal recibida no sigue una relación lineal con la posición del blanco. Muchas de las soluciones actuales al problema de localization usando RSSI se basan en complejos esquemas centralizados como filtros de partículas, mientas que en otras se basan en esquemas mucho más simples pero con menor precisión. Además, en muchos casos las estrategias son centralizadas lo que resulta poco prácticos para su implementación en redes de sensores. Desde un punto de vista práctico y de implementation, es conveniente, para ciertos escenarios y aplicaciones, el desarrollo de alternativas que ofrezcan un compromiso entre complejidad y precisión. En esta línea, en lugar de abordar directamente el problema de la estimación de la posición del blanco bajo el criterio de máxima verosimilitud, proponemos usar una formulación subóptima del problema más manejable analíticamente y que ofrece la ventaja de permitir en¬contrar la solución al problema de localization de una forma totalmente distribuida, convirtiéndola así en una solución atractiva dentro del contexto de redes inalámbricas de sensores. Para ello, se usan herramientas de procesado distribuido como los algorit¬mos de consenso y de optimización convexa en sistemas distribuidos. Para aplicaciones donde se requiera de un mayor grado de precisión se propone una estrategia que con¬siste en la optimización local de la función de verosimilitud entorno a la estimación inicialmente obtenida. Esta optimización se puede realizar de forma descentralizada usando una versión basada en consenso del método de Gauss-Newton siempre y cuando asumamos independencia de los ruidos de medida en los diferentes nodos. Independientemente de la aplicación subyacente de la red de sensores, es necesario tener un mecanismo que permita recopilar los datos provenientes de la red de sensores. Una forma de hacerlo es mediante el uso de uno o varios nodos especiales, llamados nodos “sumidero”, (sink en inglés) que actúen como centros recolectores de información y que estarán equipados con hardware adicional que les permita la interacción con el exterior de la red. La principal desventaja de esta estrategia es que dichos nodos se convierten en cuellos de botella en cuanto a tráfico y capacidad de cálculo. Como alter¬nativa se pueden usar técnicas cooperativas de conformación de haz (beamforming en inglés) de manera que el conjunto de la red puede verse como un único sistema virtual de múltiples antenas y, por tanto, que exploten los beneficios que ofrecen las comu¬nicaciones con múltiples antenas. Para ello, los distintos nodos de la red sincronizan sus transmisiones de manera que se produce una interferencia constructiva en el recep¬tor. No obstante, las actuales técnicas se basan en resultados promedios y asintóticos, cuando el número de nodos es muy grande. Para una configuración específica se pierde el control sobre el diagrama de radiación causando posibles interferencias sobre sis¬temas coexistentes o gastando más potencia de la requerida. La eficiencia energética es una cuestión capital en las redes inalámbricas de sensores ya que los nodos están equipados con baterías. Es por tanto muy importante preservar la batería evitando cambios innecesarios y el consecuente aumento de costes. Bajo estas consideraciones, se propone un esquema de conformación de haz que maximice el tiempo de vida útil de la red, entendiendo como tal el máximo tiempo que la red puede estar operativa garantizando unos requisitos de calidad de servicio (QoS por sus siglas en inglés) que permitan una decodificación fiable de la señal recibida en la estación base. Se proponen además algoritmos distribuidos que convergen a la solución centralizada. Inicialmente se considera que la única causa de consumo energético se debe a las comunicaciones con la estación base. Este modelo de consumo energético es modificado para tener en cuenta otras formas de consumo de energía derivadas de procesos inherentes al funcionamiento de la red como la adquisición y procesado de datos, las comunicaciones locales entre nodos, etc. Dicho consumo adicional de energía se modela como una variable aleatoria en cada nodo. Se cambia por tanto, a un escenario probabilístico que generaliza el caso determinista y se proporcionan condiciones bajo las cuales el problema se puede resolver de forma eficiente. Se demuestra que el tiempo de vida de la red mejora de forma significativa usando el criterio propuesto de eficiencia energética.

Computational methods to create and analyze a digital gene expression atlas of embryo development from microscopy images

Abstract The creation of atlases, or digital models where information from different subjects can be combined, is a field of increasing interest in biomedical imaging. When a single image does not contain enough information to appropriately describe the organism under study, it is then necessary to acquire images of several individuals, each of them containing complementary data with respect to the rest of the components in the cohort. This approach allows creating digital prototypes, ranging from anatomical atlases of human patients and organs, obtained for instance from Magnetic Resonance Imaging, to gene expression cartographies of embryo development, typically achieved from Light Microscopy. Within such context, in this PhD Thesis we propose, develop and validate new dedicated image processing methodologies that, based on image registration techniques, bring information from multiple individuals into alignment within a single digital atlas model. We also elaborate a dedicated software visualization platform to explore the resulting wealth of multi-dimensional data and novel analysis algo-rithms to automatically mine the generated resource in search of bio¬logical insights. In particular, this work focuses on gene expression data from developing zebrafish embryos imaged at the cellular resolution level with Two-Photon Laser Scanning Microscopy. Disposing of quantitative measurements relating multiple gene expressions to cell position and their evolution in time is a fundamental prerequisite to understand embryogenesis multi-scale processes. However, the number of gene expressions that can be simultaneously stained in one acquisition is limited due to optical and labeling constraints. These limitations motivate the implementation of atlasing strategies that can recreate a virtual gene expression multiplex. The developed computational tools have been tested in two different scenarios. The first one is the early zebrafish embryogenesis where the resulting atlas constitutes a link between the phenotype and the genotype at the cellular level. The second one is the late zebrafish brain where the resulting atlas allows studies relating gene expression to brain regionalization and neurogenesis. The proposed computational frameworks have been adapted to the requirements of both scenarios, such as the integration of partial views of the embryo into a whole embryo model with cellular resolution or the registration of anatom¬ical traits with deformable transformation models non-dependent on any specific labeling. The software implementation of the atlas generation tool (Match-IT) and the visualization platform (Atlas-IT) together with the gene expression atlas resources developed in this Thesis are to be made freely available to the scientific community. Lastly, a novel proof-of-concept experiment integrates for the first time 3D gene expression atlas resources with cell lineages extracted from live embryos, opening up the door to correlate genetic and cellular spatio-temporal dynamics. La creación de atlas, o modelos digitales, donde la información de distintos sujetos puede ser combinada, es un campo de creciente interés en imagen biomédica. Cuando una sola imagen no contiene suficientes datos como para describir apropiadamente el organismo objeto de estudio, se hace necesario adquirir imágenes de varios individuos, cada una de las cuales contiene información complementaria respecto al resto de componentes del grupo. De este modo, es posible crear prototipos digitales, que pueden ir desde atlas anatómicos de órganos y pacientes humanos, adquiridos por ejemplo mediante Resonancia Magnética, hasta cartografías de la expresión genética del desarrollo de embrionario, típicamente adquiridas mediante Microscopía Optica. Dentro de este contexto, en esta Tesis Doctoral se introducen, desarrollan y validan nuevos métodos de procesado de imagen que, basándose en técnicas de registro de imagen, son capaces de alinear imágenes y datos provenientes de múltiples individuos en un solo atlas digital. Además, se ha elaborado una plataforma de visualization específicamente diseñada para explorar la gran cantidad de datos, caracterizados por su multi-dimensionalidad, que resulta de estos métodos. Asimismo, se han propuesto novedosos algoritmos de análisis y minería de datos que permiten inspeccionar automáticamente los atlas generados en busca de conclusiones biológicas significativas. En particular, este trabajo se centra en datos de expresión genética del desarrollo embrionario del pez cebra, adquiridos mediante Microscopía dos fotones con resolución celular. Disponer de medidas cuantitativas que relacionen estas expresiones genéticas con las posiciones celulares y su evolución en el tiempo es un prerrequisito fundamental para comprender los procesos multi-escala característicos de la morfogénesis. Sin embargo, el número de expresiones genéticos que pueden ser simultáneamente etiquetados en una sola adquisición es reducido debido a limitaciones tanto ópticas como del etiquetado. Estas limitaciones requieren la implementación de estrategias de creación de atlas que puedan recrear un multiplexado virtual de expresiones genéticas. Las herramientas computacionales desarrolladas han sido validadas en dos escenarios distintos. El primer escenario es el desarrollo embrionario temprano del pez cebra, donde el atlas resultante permite constituir un vínculo, a nivel celular, entre el fenotipo y el genotipo de este organismo modelo. El segundo escenario corresponde a estadios tardíos del desarrollo del cerebro del pez cebra, donde el atlas resultante permite relacionar expresiones genéticas con la regionalización del cerebro y la formación de neuronas. La plataforma computacional desarrollada ha sido adaptada a los requisitos y retos planteados en ambos escenarios, como la integración, a resolución celular, de vistas parciales dentro de un modelo consistente en un embrión completo, o el alineamiento entre estructuras de referencia anatómica equivalentes, logrado mediante el uso de modelos de transformación deformables que no requieren ningún marcador específico. Está previsto poner a disposición de la comunidad científica tanto la herramienta de generación de atlas (Match-IT), como su plataforma de visualización (Atlas-IT), así como las bases de datos de expresión genética creadas a partir de estas herramientas. Por último, dentro de la presente Tesis Doctoral, se ha incluido una prueba conceptual innovadora que permite integrar los mencionados atlas de expresión genética tridimensionales dentro del linaje celular extraído de una adquisición in vivo de un embrión. Esta prueba conceptual abre la puerta a la posibilidad de correlar, por primera vez, las dinámicas espacio-temporales de genes y células.

3D + t Morphological Processing: Applications to Embryogenesis Image Analysis

We propose to directly process 3D + t image sequences with mathematical morphology operators, using a new classi?cation of the 3D+t structuring elements. Several methods (?ltering, tracking, segmentation) dedicated to the analysis of 3D + t datasets of zebra?sh embryogenesis are introduced and validated through a synthetic dataset. Then, we illustrate the application of these methods to the analysis of datasets of zebra?sh early development acquired with various microscopy techniques. This processing paradigm produces spatio-temporal coherent results as it bene?ts from the intrinsic redundancy of the temporal dimension, and minimizes the needs for human intervention in semi-automatic algorithms.

Assembling models of embryo development: Image analysis and the construction of digital atlases

Digital atlases of animal development provide a quantitative description of morphogenesis, opening the path toward processes modeling. Prototypic atlases offer a data integration framework where to gather information from cohorts of individuals with phenotypic variability. Relevant information for further theoretical reconstruction includes measurements in time and space for cell behaviors and gene expression. The latter as well as data integration in a prototypic model, rely on image processing strategies. Developing the tools to integrate and analyze biological multidimensional data are highly relevant for assessing chemical toxicity or performing drugs preclinical testing. This article surveys some of the most prominent efforts to assemble these prototypes, categorizes them according to salient criteria and discusses the key questions in the field and the future challenges toward the reconstruction of multiscale dynamics in model organisms.