A watershed and active contours based method for dendritic spine segmentation in 2-photon microscopy images (2-Foton mikroskopi görüntülerindeki dendritik dikenlerin bölütlenmesi için watershed ve etkin çevritlere dayalı bir yöntem)

Analysing morphological and volumetric properties of dendritic spines from 2-photon microscopy images has been of interest to neuroscientists in recent years. Developing robust and reliable tools for automatic analysis depends on the segmentation quality. In this paper, we propose a new segmentation algorithm for dendritic spine segmentation based on watershed and active contour methods. First, our proposed method coarsely segments the dendritic spine area using the watershed algorithm. Then, these results are further refined using a region-based active contour approach. We compare our results and the results of existing methods in the literature to manual delineations of a domain expert. Experimental results demonstrate that our proposed method produces more accurate results than the existing algorithms proposed for dendritic spine segmentation

Automatic dendritic spine detection using multiscale dot enhancement filters and sift features

Statistical characterization of morphological changes of dendritic spines is becoming of crucial interest in the field of neurobiology. Automatic detection and segmentation of dendritic spines promises significant reductions on the time spent by the scientists and reduces the subjectivity concerns. In this paper, we present two approaches for automated detection of dendritic spines in 2-photon laser scanning microscopy (2pLSM) images. The first method combines the idea of dot enhancement filters with information from the dendritic skeleton. The second method learns an SVM classifier by utilizing some pre-labeled SIFT feature descriptors and uses the classifier to detect dendritic spines in new images. For the segmentation of detected spines, we employ a watershed-variational segmentation algorithm. We evaluate the proposed approaches by comparing with manual segmentations of domain experts and the results of a noncommercial software, NeuronIQ. Our methods produce promising detection rate with high segmentation accuracy thus can serve as a useful tool for spine analysis

Image informatics strategies for deciphering neuronal network connectivity

Brain function relies on an intricate network of highly dynamic neuronal connections that rewires dramatically under the impulse of various external cues and pathological conditions. Among the neuronal structures that show morphologi- cal plasticity are neurites, synapses, dendritic spines and even nuclei. This structural remodelling is directly connected with functional changes such as intercellular com- munication and the associated calcium-bursting behaviour. In vitro cultured neu- ronal networks are valuable models for studying these morpho-functional changes. Owing to the automation and standardisation of both image acquisition and image analysis, it has become possible to extract statistically relevant readout from such networks. Here, we focus on the current state-of-the-art in image informatics that enables quantitative microscopic interrogation of neuronal networks. We describe the major correlates of neuronal connectivity and present workflows for analysing them. Finally, we provide an outlook on the challenges that remain to be addressed, and discuss how imaging algorithms can be extended beyond in vitro imaging studies

3D dendritic spine segmentation using nonparametric shape priors (3B dendritik dikenlerin parametrik olmayan şekil ön bilgisi kullanılarak bölütlenmesi)

Analyzing morphological and structural changes of dendritic spines in 2-photon microscopy images in time is important for neuroscience researchers. Correct segmentation of dendritic spines is an important step of developing robust and reliable automatic tools for such analysis. In this paper, we propose an approach for segmentation of 3D dendritic spines using nonparametric shape priors. The proposed method learns the prior distribution of shapes through Parzen density estimation on the training set of shapes. Then, the posterior distribution of shapes is obtained by combining the learned prior distribution with a data term in a Bayesian framework. Finally, the segmentation result that maximizes the posterior is found using active contours. Experimental results demonstrate that using nonparametric shape priors leads to better 3D dendritic spine segmentation results

Three-dimensional reengineering of neuronal microcircuits : The cortical column in silico

The presented thesis will describe a pipeline to reengineer three-dimensional, anatomically realistic, functional neuronal networks with subcellular resolution. The pipeline consists of five methods: 1. "NeuroCount" provides the number and three-dimensional distribution of all neuron somata in large brain regions. 2. "NeuroMorph" provides authentic neuron tracings, comprising dendrite and axon morphology. 3. "daVinci" registers the neuron morphologies to a standardized reference framework. 4. "NeuroCluster" objectively groups the standardized tracings into anatomical neuron types. 5. "NeuroNet" combines the number and distribution of neurons and neuron-types with the standardized tracings and determines the neuron-type- and position-specific number of synaptic connections for any two types of neuron. The developed methods are demonstrated by reengineering the thalamocortical lemniscal microcircuit in the somatosensory system of rats. There exists an one-to-one correspondence between the sensory information obtained by a single facial whisker and segregated areas in the thalamus and cortex. The reengineering of this pathway results in a column-shaped network model of ~15200 excitatory full-compartmental cortical neurons. This network is synaptically connected to ~285 pre-synaptic thalamic neurons. Animation of this "cortical column in silico" with measured physiological input will help to gain a mechanistic understanding of neuronal sensory information processing in the mammalian brain

Dendritic spine shape analysis based on two-photon microscopy images

Neuronal morphology and function are highly coupled. In particular, dendritic spine morphology is strongly governed by the incoming neuronal activity. Previously, volumes of dendritic spines have been considered as a primary parameter to study spine morphology and gain insight into structure-function coupling. However, this reductionist approach fails to incorporate the broad spine structure repertoire. First step towards integrating the rich spine morphology information into functional coupling is to classify spine shapes into main spine types suggested in the literature. Due to the lack of reliable automated analysis tools, classification is currently performed manually, which is a time-intensive task and prone to subjectivity. Availability of automated spine shape analysis tools can accelerate this process and help neuroscientists understand underlying structure and function relationship. Several studies on spine shape classification have been reported in the literature, however, there is an on-going debate on whether distinct spine shape classes exist or whether spines should be modeled through a continuum of shape variations. Another challenge is the subjectivity and bias that is introduced due to the supervised nature of classification approaches. This thesis focuses on morphological, shape, and appearance features based methods to perform dendritic spine shape analysis using both clustering and classification approaches. We apply manifold learning methods for dendritic spine classification and observe that ISOMAP implicitly computes prominent features suitable for classification purposes. We also apply linear representation based approach for spine classification and conclude that sparse representation provides slightly better classification performance. We propose 2D and 3D morphological features based approach for spine shape analysis and demonstrate the advantage of 3D morphological features. We also use a deep learning based approach for spine classification and show that mid-level features extracted from Convolutional Neural Networks (CNNs) perform as well as hand-crafted features. We propose a kernel density estimation (KDE) based framework for dendritic spine classification. We evaluate our proposed approaches by comparing labels assigned by a neuroscience expert. Our KDE based framework also enables neuroscientists to analyze separability of spine shape classes in the likelihood ratio space, which leads to further insights about the nature of the spine shape analysis problem. Furthermore, we also propose a methodology for unsupervised learning and clustering of spine shapes. In particular, we use x-means to perform cluster analysis that selects the number of clusters automatically using the Bayesian information criterion (BIC). The objective of clustering in this context is two-fold: confirm the hypothesis of some distinct shape classes and discover new natural groups. We observe that although there are many spines which easily fit into the definition of standard shape types (confirming the hypothesis), there are also a significant number of others which do not comply with standard shape types and demonstrate intermediate properties

Doctor of Philosophy

dissertationNeuroscientists are developing new imaging techniques and generating large volumes of data in an effort to understand the complex structure of the nervous system. The complexity and size of this data makes human interpretation a labor intensive task. To aid in the analysis, new segmentation techniques for identifying neurons in these feature rich datasets are required. However, the extremely anisotropic resolution of the data makes segmentation and tracking across slices difficult. Furthermore, the thickness of the slices can make the membranes of the neurons hard to identify. Similarly, structures can change significantly from one section to the next due to slice thickness which makes tracking difficult. This thesis presents a complete method for segmenting many neurons at once in two-dimensional (2D) electron microscopy images and reconstructing and visualizing them in three-dimensions (3D). First, we present an advanced method for identifying neuron membranes in 2D, necessary for whole neuron segmentation, using a machine learning approach. The method described uses a series of artificial neural networks (ANNs) in a framework combined with a feature vector that is composed of image and context; intensities sampled over a stencil neighborhood. Several ANNs are applied in series allowing each ANN to use the classification context; provided by the previous network to improve detection accuracy. To improve the membrane detection, we use information from a nonlinear alignment of sequential learned membrane images in a final ANN that improves membrane detection in each section. The final output, the detected membranes, are used to obtain 2D segmentations of all the neurons in an image. We also present a method that constructs 3D neuron representations by formulating the problem of finding paths through sets of sections as an optimal path computation, which applies a cost function to the identification of a cell from one section to the next and solves this optimization problem using Dijkstras algorithm. This basic formulation accounts for variability or inconsistencies between sections and prioritizes cells based on the evidence of their connectivity. Finally, we present a tool that combines these techniques with a visual user interface that enables users to quickly segment whole neurons in large volumes

Enabling Scalable Neurocartography: Images to Graphs for Discovery

In recent years, advances in technology have enabled researchers to ask new questions predicated on the collection and analysis of big datasets that were previously too large to study. More specifically, many fundamental questions in neuroscience require studying brain tissue at a large scale to discover emergent properties of neural computation, consciousness, and etiologies of brain disorders. A major challenge is to construct larger, more detailed maps (e.g., structural wiring diagrams) of the brain, known as connectomes. Although raw data exist, obstacles remain in both algorithm development and scalable image analysis to enable access to the knowledge within these data volumes. This dissertation develops, combines and tests state-of-the-art algorithms to estimate graphs and glean other knowledge across six orders of magnitude, from millimeter-scale magnetic resonance imaging to nanometer-scale electron microscopy. This work enables scientific discovery across the community and contributes to the tools and services offered by NeuroData and the Open Connectome Project. Contributions include creating, optimizing and evaluating the first known fully-automated brain graphs in electron microscopy data and magnetic resonance imaging data; pioneering approaches to generate knowledge from X-Ray tomography imaging; and identifying and solving a variety of image analysis challenges associated with building graphs suitable for discovery. These methods were applied across diverse datasets to answer questions at scales not previously explored

A model-based method for 3D reconstruction of cerebellar parallel fibres from high-resolution electron microscope images

In order to understand how the brain works, we need to understand how its neural circuits process information. Electron microscopy remains the only imaging technique capable of providing sufficient resolution to reconstruct the dense connectivity between all neurons in a circuit. Automated electron microscopy techniques are approaching the point where usefully large circuits might be successfully imaged, but the development of automated reconstruction techniques lags far behind. No fully-automated reconstruction technique currently produces acceptably accurate reconstructions, and semi-automated approaches currently require an extreme amount of manual effort. This reconstruction bottleneck places severe limits on the size of neural circuits that can be reconstructed. Improved automated reconstruction techniques are therefore highly desired and under active development. The human brain contains ~86 billion neurons and ~80% of these are located in the cerebellum. Of these cerebellar neurons, the vast majority are granule cells. The axons of these granule cells are called parallel fibres and tend to be oriented in approximately the same direction, making 2+1D reconstruction approaches feasible. In this work we focus on the problem of reconstructing these parallel fibres and make four main contributions: (1) a model-based algorithm for reconstructing 2D parallel fibre cross-sections that achieves state of the art 2D reconstruction performance; (2) a fully-automated algorithm for reconstructing 3D parallel fibres that achieves state of the art 3D reconstruction performance; (3) a semi-automated approach for reconstructing 3D parallel fibres that significantly improves reconstruction accuracy compared to our fully-automated approach while requiring ~40 times less labelling effort than a purely manual reconstruction; (4) a "gold standard" ground truth data set for the molecular layer of the mouse cerebellum that will provide a valuable reference for the development and benchmarking of reconstruction algorithms