399 research outputs found
Annotating Synapses in Large EM Datasets
Reconstructing neuronal circuits at the level of synapses is a central
problem in neuroscience and becoming a focus of the emerging field of
connectomics. To date, electron microscopy (EM) is the most proven technique
for identifying and quantifying synaptic connections. As advances in EM make
acquiring larger datasets possible, subsequent manual synapse identification
({\em i.e.}, proofreading) for deciphering a connectome becomes a major time
bottleneck. Here we introduce a large-scale, high-throughput, and
semi-automated methodology to efficiently identify synapses. We successfully
applied our methodology to the Drosophila medulla optic lobe, annotating many
more synapses than previous connectome efforts. Our approaches are extensible
and will make the often complicated process of synapse identification
accessible to a wider-community of potential proofreaders
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
Synaptic Cleft Segmentation in Non-Isotropic Volume Electron Microscopy of the Complete Drosophila Brain
Neural circuit reconstruction at single synapse resolution is increasingly
recognized as crucially important to decipher the function of biological
nervous systems. Volume electron microscopy in serial transmission or scanning
mode has been demonstrated to provide the necessary resolution to segment or
trace all neurites and to annotate all synaptic connections.
Automatic annotation of synaptic connections has been done successfully in
near isotropic electron microscopy of vertebrate model organisms. Results on
non-isotropic data in insect models, however, are not yet on par with human
annotation.
We designed a new 3D-U-Net architecture to optimally represent isotropic
fields of view in non-isotropic data. We used regression on a signed distance
transform of manually annotated synaptic clefts of the CREMI challenge dataset
to train this model and observed significant improvement over the state of the
art.
We developed open source software for optimized parallel prediction on very
large volumetric datasets and applied our model to predict synaptic clefts in a
50 tera-voxels dataset of the complete Drosophila brain. Our model generalizes
well to areas far away from where training data was available
TrakEM2 Software for Neural Circuit Reconstruction
A key challenge in neuroscience is the expeditious reconstruction of neuronal circuits. For model systems such as Drosophila and C. elegans, the limiting step is no longer the acquisition of imagery but the extraction of the circuit from images. For this purpose, we designed a software application, TrakEM2, that addresses the systematic reconstruction of neuronal circuits from large electron microscopical and optical image volumes. We address the challenges of image volume composition from individual, deformed images; of the reconstruction of neuronal arbors and annotation of synapses with fast manual and semi-automatic methods; and the management of large collections of both images and annotations. The output is a neural circuit of 3d arbors and synapses, encoded in NeuroML and other formats, ready for analysis
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