Segment and track neurons in 3D by repulsive snake method

We present a snake (active contour) model based on repulsive force to segment neurons obtained from microscopy. Based on these segmentation results, we track the neurons in 3D image to look for its branch structure. These segmentation results allow user to study morphology of neurons to further investigate neuronal function and connectivity. This repulsive snake model can successfully segment two or multiple neurons that are close to each other by some alternating repulsive force generated from the neighboring objects. We apply our results on real data to demonstrate the performance of our method. © 2005 IEEE.published_or_final_versio

Model and Appearance Based Analysis of Neuronal Morphology from Different Microscopy Imaging Modalities

The neuronal morphology analysis is key for understanding how a brain works. This process requires the neuron imaging system with single-cell resolution; however, there is no feasible system for the human brain. Fortunately, the knowledge can be inferred from the model organism, Drosophila melanogaster, to the human system. This dissertation explores the morphology analysis of Drosophila larvae at single-cell resolution in static images and image sequences, as well as multiple microscopy imaging modalities. Our contributions are on both computational methods for morphology quantification and analysis of the influence of the anatomical aspect. We develop novel model-and-appearance-based methods for morphology quantification and illustrate their significance in three neuroscience studies. Modeling of the structure and dynamics of neuronal circuits creates understanding about how connectivity patterns are formed within a motor circuit and determining whether the connectivity map of neurons can be deduced by estimations of neuronal morphology. To address this problem, we study both boundary-based and centerline-based approaches for neuron reconstruction in static volumes. Neuronal mechanisms are related to the morphology dynamics; so the patterns of neuronal morphology changes are analyzed along with other aspects. In this case, the relationship between neuronal activity and morphology dynamics is explored to analyze locomotion procedures. Our tracking method models the morphology dynamics in the calcium image sequence designed for detecting neuronal activity. It follows the local-to-global design to handle calcium imaging issues and neuronal movement characteristics. Lastly, modeling the link between structural and functional development depicts the correlation between neuron growth and protein interactions. This requires the morphology analysis of different imaging modalities. It can be solved using the part-wise volume segmentation with artificial templates, the standardized representation of neurons. Our method follows the global-to-local approach to solve both part-wise segmentation and registration across modalities. Our methods address common issues in automated morphology analysis from extracting morphological features to tracking neurons, as well as mapping neurons across imaging modalities. The quantitative analysis delivered by our techniques enables a number of new applications and visualizations for advancing the investigation of phenomena in the nervous system

Semi-Automated Reconstruction of Neural Processes from Large Numbers of Fluorescence Images

We introduce a method for large scale reconstruction of complex bundles of neural processes from fluorescent image stacks. We imaged yellow fluorescent protein labeled axons that innervated a whole muscle, as well as dendrites in cerebral cortex, in transgenic mice, at the diffraction limit with a confocal microscope. Each image stack was digitally re-sampled along an orientation such that the majority of axons appeared in cross-section. A region growing algorithm was implemented in the open-source Reconstruct software and applied to the semi-automatic tracing of individual axons in three dimensions. The progression of region growing is constrained by user-specified criteria based on pixel values and object sizes, and the user has full control over the segmentation process. A full montage of reconstructed axons was assembled from the ∼200 individually reconstructed stacks. Average reconstruction speed is ∼0.5 mm per hour. We found an error rate in the automatic tracing mode of ∼1 error per 250 um of axonal length. We demonstrated the capacity of the program by reconstructing the connectome of motor axons in a small mouse muscle

Automatic segmentation of overlapping cervical smear cells based on local distinctive features and guided shape deformation

Automated segmentation of cells from cervical smears poses great challenge to biomedical image analysis because of the noisy and complex background, poor cytoplasmic contrast and the presence of fuzzy and overlapping cells. In this paper, we propose an automated segmentation method for the nucleus and cytoplasm in a cluster of cervical cells based on distinctive local features and guided sparse shape deformation. Our proposed approach is performed in two stages: segmentation of nuclei and cellular clusters, and segmentation of overlapping cytoplasm. In the rst stage, a set of local discriminative shape and appearance cues of image superpixels is incorporated and classi ed by the Support Vector Machine (SVM) to segment the image into nuclei, cellular clusters, and background. In the second stage, a robust shape deformation framework is proposed, based on Sparse Coding (SC) theory and guided by representative shape features, to construct the cytoplasmic shape of each overlapping cell. Then, the obtained shape is re ned by the Distance Regularized Level Set Evolution (DRLSE) model. We evaluated our approach using the ISBI 2014 challenge dataset, which has 135 synthetic cell images for a total of 810 cells. Our results show that our approach outperformed existing approaches in segmenting overlapping cells and obtaining accurate nuclear boundaries. Keywords: overlapping cervical smear cells, feature extraction, sparse coding, shape deformation, distance regularized level set

A study on morphological and dynamical properties of neuronal growth cones

In the developing nervous system and in the adult brain, neurons constantly need to solve mechanical problems. Neuronal growth cones are the main motile structures located at the tip of neurites and are composed of a lamellipodium from which thin filopodia emerge. They are responsible for extension of neurite processes and for transducing signals from extracellular cues to alter directionality, branching, and motility. They must decide how to explore the environment and in which direction to grow; they also need to establish appropriate contacts, to avoid obstacles and to determine how much force to exert. The complete understanding of the nervous system and its basic unit, the neuron, demands a quantification of the behavioral pattern and the morphological characteristics..

AUTOMATED ANALYSIS OF NEURONAL MORPHOLOGY: DETECTION, MODELING AND RECONSTRUCTION

Ph.DDOCTOR OF PHILOSOPH

Methods for Automated Neuron Image Analysis

Knowledge of neuronal cell morphology is essential for performing specialized analyses in the endeavor to understand neuron behavior and unravel the underlying principles of brain function. Neurons can be captured with a high level of detail using modern microscopes, but many neuroscientific studies require a more explicit and accessible representation than offered by the resulting images, underscoring the need for digital reconstruction of neuronal morphology from the images into a tree-like graph structure. This thesis proposes new computational methods for automated detection and reconstruction of neurons from fluorescence microscopy images. Specifically, the successive chapters describe and evaluate original solutions to problems such as the detection of landmarks (critical points) of the neuronal tree, complete tracing and reconstruction of the tree, and the detection of regions containing neurons in high-content screens