35 research outputs found
Focused Proofreading: Efficiently Extracting Connectomes from Segmented EM Images
Identifying complex neural circuitry from electron microscopic (EM) images
may help unlock the mysteries of the brain. However, identifying this circuitry
requires time-consuming, manual tracing (proofreading) due to the size and
intricacy of these image datasets, thus limiting state-of-the-art analysis to
very small brain regions. Potential avenues to improve scalability include
automatic image segmentation and crowd sourcing, but current efforts have had
limited success. In this paper, we propose a new strategy, focused
proofreading, that works with automatic segmentation and aims to limit
proofreading to the regions of a dataset that are most impactful to the
resulting circuit. We then introduce a novel workflow, which exploits
biological information such as synapses, and apply it to a large dataset in the
fly optic lobe. With our techniques, we achieve significant tracing speedups of
3-5x without sacrificing the quality of the resulting circuit. Furthermore, our
methodology makes the task of proofreading much more accessible and hence
potentially enhances the effectiveness of crowd sourcing
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Neural Process Reconstruction from Sparse User Scribbles
We present a novel semi-automatic method for segmenting neural processes in large, highly anisotropic EM (electron microscopy) image stacks. Our method takes advantage of sparse scribble annotations provided by the user to guide a 3D variational segmentation model, thereby allowing our method to globally optimally enforce 3D geometric constraints on the segmentation. Moreover, we leverage a novel algorithm for propagating segmentation constraints through the image stack via optimal volumetric pathways, thereby allowing our method to compute highly accurate 3D segmentations from very sparse user input. We evaluate our method by reconstructing 16 neural processes in a 1024×1024×50 nanometer-scale EM image stack of a mouse hippocampus. We demonstrate that, on average, our method is 68% more accurate than previous state-of-the-art semi-automatic methods.Engineering and Applied Science
A circle-based method for detection of neural fibre cross-sections in classically stained 2D electron micrographs
Recent developments in electron microscopy now permit the unambiguous reconstruction of even the smallest neural fibres by human experts. However, manual reconstruction of an interesting volume of neural tissue would take thousands of person-years. Techniques to automate such reconstruction are therefore highly desirable and currently under active development. Here we present a novel circle-based technique and assess its performance on classically stained electron micrographs of the molecular layer of mouse cerebellar cortex. We compare its performance to a recently published pixel-based classifier (ilastik), selected because a similar random forest classifier from the same group has shown promising results on images of neural tissue. The performance of our algorithm and that of ilastik are similar, achieving approximately 50% on an overlap-based f-measure
Automated Detection and Segmentation of Synaptic Contacts in Nearly Isotropic Serial Electron Microscopy Images
We describe a protocol for fully automated detection and segmentation of asymmetric, presumed excitatory, synapses in serial electron microscopy images of the adult mammalian cerebral cortex, taken with the focused ion beam, scanning electron microscope (FIB/SEM). The procedure is based on interactive machine learning and only requires a few labeled synapses for training. The statistical learning is performed on geometrical features of 3D neighborhoods of each voxel and can fully exploit the high z-resolution of the data. On a quantitative validation dataset of 111 synapses in 409 images of 1948×1342 pixels with manual annotations by three independent experts the error rate of the algorithm was found to be comparable to that of the experts (0.92 recall at 0.89 precision). Our software offers a convenient interface for labeling the training data and the possibility to visualize and proofread the results in 3D. The source code, the test dataset and the ground truth annotation are freely available on the website http://www.ilastik.org/synapse-detection
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Automated computation of arbor densities: a step toward identifying neuronal cell types
The shape and position of a neuron convey information regarding its molecular and functional identity. The identification of cell types from structure, a classic method, relies on the time-consuming step of arbor tracing. However, as genetic tools and imaging methods make data-driven approaches to neuronal circuit analysis feasible, the need for automated processing increases. Here, we first establish that mouse retinal ganglion cell types can be as precise about distributing their arbor volumes across the inner plexiform layer as they are about distributing the skeletons of the arbors. Then, we describe an automated approach to computing the spatial distribution of the dendritic arbors, or arbor density, with respect to a global depth coordinate based on this observation. Our method involves three-dimensional reconstruction of neuronal arbors by a supervised machine learning algorithm, post-processing of the enhanced stacks to remove somata and isolate the neuron of interest, and registration of neurons to each other using automatically detected arbors of the starburst amacrine interneurons as fiducial markers. In principle, this method could be generalizable to other structures of the CNS, provided that they allow sparse labeling of the cells and contain a reliable axis of spatial reference