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

Precise segmentation of densely interweaving neuron clusters using G-Cut

脑是宇宙间最为复杂的系统之一，成人的脑中有约1000亿个神经元，单个神经元通常与其它神经元有成千上万个“突触”连接节点，形成拥有百万亿级连接的极其复杂的脑神经网络。当前多数神经元三维重建和分析工具仅适用于单个神经元的形态学重建，难以从神经元簇图像中正确追踪重建出多个神经元，而神经元的重建质量又影响到量化分析神经元的形态学特征及其功能。针对这一问题，课题组提出一种新的三维神经元簇重建工具G-Cut。具体地，为了度量神经元胞体与神经突起间的关联性，课题组从已有的带有标注的大规模神经元形态学数据集统计分析得到其规律和形态学信息。然后将神经元簇的重建问题转化为神经突起之间连接所形成的拓扑连接图的图分割问题，并结合神经元形态学规律和信息，在所有的神经突起与神经元胞体的关联性中寻找重建问题的最优解。通过在不同的合成数据集以及真实的脑组织图像数据集上测试，和已有的方法相比，G-Cut在不同密度和不同规模的神经元簇图像上均获得了更高的重建正确率。该项研究工作由厦门大学，南加州大学，加州大学洛杉矶分校等高校课题组合作完成，厦门大学信息学院智能科学与技术系为第一完成单位，厦门大学博士生李睿和USC博士生Muye Zhu为论文共同第一作者，张俊松博士和南加州大学的Hong-Wei Dong教授为论文共同通讯作者。厦门大学周昌乐教授和南加州大学的Arthur Toga教授为研究提供了大力支持。【Abstract】Characterizing the precise three-dimensional morphology and anatomical context of neurons is crucial for neuronal cell type classification and circuitry mapping. Recent advances in tissue clearing techniques and microscopy make it possible to obtain image stacks of intact, interweaving neuron clusters in brain tissues. As most current 3D neuronal morphology reconstruction methods are only applicable to single neurons, it remains challenging to reconstruct these clusters digitally. To advance the state of the art beyond these challenges, we propose a fast and robust method named G-Cut that is able to automatically segment individual neurons from an interweaving neuron cluster. Across various densely interconnected neuron clusters, G-Cut achieves significantly higher accuracies than other state-of-the-art algorithms. G-Cut is intended as a robust component in a high throughput informatics pipeline for large-scale brain mapping projects.This work was supported by NIH/NIMH MH094360-01A1 (H.W.D.), MH094360-06 (H.W.D.), NIH/NCI U01CA198932-01 (H.W.D.), NIH/NIMH MH106008 (X.W.Y. and H.W.D.), National Nature Science Foundation of China No. 61772440 (J.S.Z.), and National Basic Research Program of China 2013CB329502 (J.S.Z. and C.L.Z.). We thank a support of Graduate Student International Exchange Project of Xiamen University to R.L. and State Scholarship Fund of China Scholarship Council (No. 201406315023) to J.S.Z. 该项研究得到国家自然科学基金、国家重点基础研究发展计划973项目、国家留学基金、厦门大学研究生国际交流项目、美国脑计划和NIH等课题资助

A workflow for the automatic segmentation of organelles in electron microscopy image stacks.

Electron microscopy (EM) facilitates analysis of the form, distribution, and functional status of key organelle systems in various pathological processes, including those associated with neurodegenerative disease. Such EM data often provide important new insights into the underlying disease mechanisms. The development of more accurate and efficient methods to quantify changes in subcellular microanatomy has already proven key to understanding the pathogenesis of Parkinson's and Alzheimer's diseases, as well as glaucoma. While our ability to acquire large volumes of 3D EM data is progressing rapidly, more advanced analysis tools are needed to assist in measuring precise three-dimensional morphologies of organelles within data sets that can include hundreds to thousands of whole cells. Although new imaging instrument throughputs can exceed teravoxels of data per day, image segmentation and analysis remain significant bottlenecks to achieving quantitative descriptions of whole cell structural organellomes. Here, we present a novel method for the automatic segmentation of organelles in 3D EM image stacks. Segmentations are generated using only 2D image information, making the method suitable for anisotropic imaging techniques such as serial block-face scanning electron microscopy (SBEM). Additionally, no assumptions about 3D organelle morphology are made, ensuring the method can be easily expanded to any number of structurally and functionally diverse organelles. Following the presentation of our algorithm, we validate its performance by assessing the segmentation accuracy of different organelle targets in an example SBEM dataset and demonstrate that it can be efficiently parallelized on supercomputing resources, resulting in a dramatic reduction in runtime

Convolutional nets for reconstructing neural circuits from brain images acquired by serial section electron microscopy

Neural circuits can be reconstructed from brain images acquired by serial section electron microscopy. Image analysis has been performed by manual labor for half a century, and efforts at automation date back almost as far. Convolutional nets were first applied to neuronal boundary detection a dozen years ago, and have now achieved impressive accuracy on clean images. Robust handling of image defects is a major outstanding challenge. Convolutional nets are also being employed for other tasks in neural circuit reconstruction: finding synapses and identifying synaptic partners, extending or pruning neuronal reconstructions, and aligning serial section images to create a 3D image stack. Computational systems are being engineered to handle petavoxel images of cubic millimeter brain volumes

Automating the Reconstruction of Neuron Morphological Models: the Rivulet Algorithm Suite

The automatic reconstruction of single neuron cells is essential to enable large-scale data-driven investigations in computational neuroscience. The problem remains an open challenge due to various imaging artefacts that are caused by the fundamental limits of light microscopic imaging. Few previous methods were able to generate satisfactory neuron reconstruction models automatically without human intervention. The manual tracing of neuron models is labour heavy and time-consuming, making the collection of large-scale neuron morphology database one of the major bottlenecks in morphological neuroscience. This thesis presents a suite of algorithms that are developed to target the challenge of automatically reconstructing neuron morphological models with minimum human intervention. We first propose the Rivulet algorithm that iteratively backtracks the neuron fibres from the termini points back to the soma centre. By refining many details of the Rivulet algorithm, we later propose the Rivulet2 algorithm which not only eliminates a few hyper-parameters but also improves the robustness against noisy images. A soma surface reconstruction method was also proposed to make the neuron models biologically plausible around the soma body. The tracing algorithms, including Rivulet and Rivulet2, normally need one or more hyper-parameters for segmenting the neuron body out of the noisy background. To make this pipeline fully automatic, we propose to use 2.5D neural network to train a model to enhance the curvilinear structures of the neuron fibres. The trained neural networks can quickly highlight the fibres of interests and suppress the noise points in the background for the neuron tracing algorithms. We evaluated the proposed methods in the data released by both the DIADEM and the BigNeuron challenge. The experimental results show that our proposed tracing algorithms achieve the state-of-the-art results