Whole-brain vasculature reconstruction at the single capillary level

The distinct organization of the brain’s vascular network ensures that it is adequately supplied with oxygen and nutrients. However, despite this fundamental role, a detailed reconstruction of the brain-wide vasculature at the capillary level remains elusive, due to insufficient image quality using the best available techniques. Here, we demonstrate a novel approach that improves vascular demarcation by combining CLARITY with a vascular staining approach that can fill the entire blood vessel lumen and imaging with light-sheet fluorescence microscopy. This method significantly improves image contrast, particularly in depth, thereby allowing reliable application of automatic segmentation algorithms, which play an increasingly important role in high-throughput imaging of the terabyte-sized datasets now routinely produced. Furthermore, our novel method is compatible with endogenous fluorescence, thus allowing simultaneous investigations of vasculature and genetically targeted neurons. We believe our new method will be valuable for future brain-wide investigations of the capillary network

Whole-brain vasculature reconstruction at the single capillary level

The distinct organization of the brain’s vascular network ensures that it is adequately supplied with oxygen and nutrients. However, despite this fundamental role, a detailed reconstruction of the brain-wide vasculature at the capillary level remains elusive, due to insufficient image quality using the best available techniques. Here, we demonstrate a novel approach that improves vascular demarcation by combining CLARITY with a vascular staining approach that can fill the entire blood vessel lumen and imaging with light-sheet fluorescence microscopy. This method significantly improves image contrast, particularly in depth, thereby allowing reliable application of automatic segmentation algorithms, which play an increasingly important role in high-throughput imaging of the terabyte-sized datasets now routinely produced. Furthermore, our novel method is compatible with endogenous fluorescence, thus allowing simultaneous investigations of vasculature and genetically targeted neurons. We believe our new method will be valuable for future brain-wide investigations of the capillary network

Doctor of Philosophy

dissertationMagnetic resonance guided high intensity focused ultrasound (MRgHIFU) is a promising minimal invasive thermal therapy for the treatment of breast cancer. This study develops techniques for determining the tissue parameters - tissue types and perfusion rate - that influence the local temperature during HIFU thermotherapy procedures. For optimal treatment planning for each individual patient, a 3D volumetric breast tissue segmentation scheme based on the hierarchical support vector machine (SVM) algorithm was developed to automatically segment breast tissues into fat, fibroglandular tissue, skin and lesions. Compared with fuzzy c-mean and conventional SVM algorithm, the presented technique offers tissue classification performance with the highest accuracy. The consistency of the segmentation results along both the sagittal and axial orientations indicates the stability of the proposed segmentation routine. Accurate knowledge of the internal anatomy of the breast can be utilized in the ultrasound beam simulation for the treatment planning of MRgHIFU therapy. Completely noninvasive MRI techniques were developed for visualizing blood vessels and determining perfusion rate to assist in the MRgHIFU therapy. Two-point Dixon fat-water separation was achieved using a 3D dual-echo SSFP sequence for breast vessel imaging. The performances of the fat-water separation with various readout gradient designs were evaluated on a water-oil phantom, ex vivo pork sample and in vivo breast imaging. Results suggested that using a dual-echo SSFP readout with bipolar readout gradient polarity, blood vasculature could be successfully visualized through the thin-slab maximum intensity projection SSFP water-only images. For determining the perfusion rate, we presented a novel imaging pulse sequence design consisting of a single arterial spin labeling (ASL) magnetization preparation followed by Look-Locker-like image readouts. This flow quantification technique was examined through simulation, in vitro and in vivo experiments. Experimental results from a hemodialyzer when fitted with a Bloch-equation-based model provide flow measurements that are consistent with ground truth velocities. With these tissue properties, it is possible to compensate for the dissipative effects of the flowing blood and ultimately improve the efficacy of the MRgHIFU therapies. Complete noninvasiveness of these techniques allows multiple measurements before, during and after the treatment, without the limitation of washout of the injected contrast agent

Tomografia estendida : do básico até o mapeamento de cérebro de camundongos

Orientador: Mateus Borba CardosoTese (doutorado) - Universidade Estadual de Campinas, Instituto de Física Gleb WataghinResumo: Esta tese apresentará uma introdução a imagens de raios-x e como adquirir e processar imagens usando linhas de luz síncrotron. Apresentará os desafios matemáticos e técnicos para reconstruir amostras em três dimensões usando a reconstrução de Tomografia Computadorizada, uma técnica conhecida como CT. Esta técnica tem seu campo de visão limitado ao tamanho da câmera e ao tamanho da iluminação. Uma técnica para ampliar esse campo de visão vai ser apresentada e os desafios técnicos envolvidos para que isso aconteça. Um \textit{pipeline} é proposto e todos os algoritmos necessários foram empacotados em um pacote python chamado Tomosaic. A abordagem baseia-se em adquirir tomogramas parciais em posiçoes pré definidas e depois mesclar os dados em um novo conjunto de dados. Duas maneiras possíveis são apresentadas para essa mescla, uma no domínio das projeções e uma no domínio dos sinogramas. Experimentos iniciais serão então usadas para mostrar que o método proposto funciona com computadores normais. A técnica será aplicada mais tarde para pesquisar a anatomia de cérebros de camundongo completos. Um estudo será apresentado de como obter informação em diferentes escalas do cérebro completo do rato utilizando raios-xAbstract: This thesis will present an introduction to x-ray images and how to acquire and thread images using synchrotron beamlines. It will present the mathematical and technical challenges to reconstruct samples in three dimensions using Computed Tomography reconstruction, a technique known as CT. This technique has a field of view bounded to the camera size and the illumination size. A technique to extended this field of view is going to be presented and the technical challenges involved in order for that to happen will be described. A pipeline is proposed and all the necessary algorithms are contained into a python packaged called Tomosaic. The approach relies on acquired partial tomogram data in a defined grid and later merging the data into a new dataset. Two possible ways are presented in order to that: in the projection domain, and in the sinogram domain. Initial experiments will then be used to show that the pipeline works with normal computers. The technique will be later applied to survey the whole anatomy of whole mouse brains. A study will be shown of how to get the complete range of scales of the mouse brain using x-ray tomography at different resolutionsDoutoradoFísicaDoutor em Ciências163304/2013-01247445/2013, 1456912/2014CNPQCAPE

Molecular Imaging

The present book gives an exceptional overview of molecular imaging. Practical approach represents the red thread through the whole book, covering at the same time detailed background information that goes very deep into molecular as well as cellular level. Ideas how molecular imaging will develop in the near future present a special delicacy. This should be of special interest as the contributors are members of leading research groups from all over the world