70 research outputs found
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The role of spatial embedding in mouse brain networks constructed from diffusion tractography and tracer injections
Diffusion MRI tractography is the only noninvasive method to measure the structural connectome in humans. However, recent validation studies have revealed limitations of modern tractography approaches, which lead to significant mistracking caused in part by local uncertainties in fiber orientations that accumulate to produce larger errors for longer streamlines. Characterizing the role of this length bias in tractography is complicated by the true underlying contribution of spatial embedding to brain topology. In this work, we compare graphs constructed with ex vivo tractography data in mice and neural tracer data from the Allen Mouse Brain Connectivity Atlas to random geometric surrogate graphs which preserve the low-order distance effects from each modality in order to quantify the role of geometry in various network properties. We find that geometry plays a substantially larger role in determining the topology of graphs produced by tractography than graphs produced by tracers. Tractography underestimates weights at long distances compared to neural tracers, which leads tractography to place network hubs close to the geometric center of the brain, as do corresponding tractography-derived random geometric surrogates, while tracer graphs place hubs further into peripheral areas of the cortex. We also explore the role of spatial embedding in modular structure, network efficiency and other topological measures in both modalities. Throughout, we compare the use of two different tractography streamline node assignment strategies and find that the overall differences between tractography approaches are small relative to the differences between tractography- and tracer-derived graphs. These analyses help quantify geometric biases inherent to tractography and promote the use of geometric benchmarking in future tractography validation efforts
Large-Scale Automatic Reconstruction of Neuronal Processes from Electron Microscopy Images
Automated sample preparation and electron microscopy enables acquisition of
very large image data sets. These technical advances are of special importance
to the field of neuroanatomy, as 3D reconstructions of neuronal processes at
the nm scale can provide new insight into the fine grained structure of the
brain. Segmentation of large-scale electron microscopy data is the main
bottleneck in the analysis of these data sets. In this paper we present a
pipeline that provides state-of-the art reconstruction performance while
scaling to data sets in the GB-TB range. First, we train a random forest
classifier on interactive sparse user annotations. The classifier output is
combined with an anisotropic smoothing prior in a Conditional Random Field
framework to generate multiple segmentation hypotheses per image. These
segmentations are then combined into geometrically consistent 3D objects by
segmentation fusion. We provide qualitative and quantitative evaluation of the
automatic segmentation and demonstrate large-scale 3D reconstructions of
neuronal processes from a volume of brain
tissue over a cube of in each dimension corresponding to
1000 consecutive image sections. We also introduce Mojo, a proofreading tool
including semi-automated correction of merge errors based on sparse user
scribbles
Distributed optimization for nonrigid nano-tomography
Resolution level and reconstruction quality in nano-computed tomography
(nano-CT) are in part limited by the stability of microscopes, because the
magnitude of mechanical vibrations during scanning becomes comparable to the
imaging resolution, and the ability of the samples to resist beam damage during
data acquisition. In such cases, there is no incentive in recovering the sample
state at different time steps like in time-resolved reconstruction methods, but
instead the goal is to retrieve a single reconstruction at the highest possible
spatial resolution and without any imaging artifacts. Here we propose a joint
solver for imaging samples at the nanoscale with projection alignment,
unwarping and regularization. Projection data consistency is regulated by dense
optical flow estimated by Farneback's algorithm, leading to sharp sample
reconstructions with less artifacts. Synthetic data tests show robustness of
the method to Poisson and low-frequency background noise. Applicability of the
method is demonstrated on two large-scale nano-imaging experimental data sets.Comment: Manuscript and supplementary materia
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Correlative light and electron microscopy using cathodoluminescence from nanoparticles with distinguishable colours
Correlative light and electron microscopy promises to combine molecular specificity with nanoscale imaging resolution. However, there are substantial technical challenges including reliable co-registration of optical and electron images, and rapid optical signal degradation under electron beam irradiation. Here, we introduce a new approach to solve these problems: imaging of stable optical cathodoluminescence emitted in a scanning electron microscope by nanoparticles with controllable surface chemistry. We demonstrate well-correlated cathodoluminescence and secondary electron images using three species of semiconductor nanoparticles that contain defects providing stable, spectrally-distinguishable cathodoluminescence. We also demonstrate reliable surface functionalization of the particles. The results pave the way for the use of such nanoparticles for targeted labeling of surfaces to provide nanoscale mapping of molecular composition, indicated by cathodoluminescence colour, simultaneously acquired with structural electron images in a single instrument.Physic
Pervasive Synaptic Branch Removal in the Mammalian Neuromuscular System at Birth
SummaryUsing light and serial electron microscopy, we show profound refinements in motor axonal branching and synaptic connectivity before and after birth. Embryonic axons become maximally connected just before birth when they innervate ∼10-fold more muscle fibers than in maturity. In some developing muscles, axons innervate almost every muscle fiber. At birth, each neuromuscular junction is coinnervated by approximately ten highly intermingled axons (versus one in adults). Extensive die off of terminal branches occurs during the first several postnatal days, leading to much sparser arbors that still span the same territory. Despite the extensive pruning, total axoplasm per neuron increases as axons elongate, thicken, and add more synaptic release sites on their remaining targets. Motor axons therefore initially establish weak connections with nearly all available postsynaptic targets but, beginning at birth, massively redistribute synaptic resources, concentrating many more synaptic sites on many fewer muscle fibers. Analogous changes in connectivity may occur in the CNS.Video Abstrac
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