298 research outputs found
A Pipeline for Volume Electron Microscopy of the Caenorhabditis elegans Nervous System.
The "connectome," a comprehensive wiring diagram of synaptic connectivity, is achieved through volume electron microscopy (vEM) analysis of an entire nervous system and all associated non-neuronal tissues. White et al. (1986) pioneered the fully manual reconstruction of a connectome using Caenorhabditis elegans. Recent advances in vEM allow mapping new C. elegans connectomes with increased throughput, and reduced subjectivity. Current vEM studies aim to not only fill the remaining gaps in the original connectome, but also address fundamental questions including how the connectome changes during development, the nature of individuality, sexual dimorphism, and how genetic and environmental factors regulate connectivity. Here we describe our current vEM pipeline and projected improvements for the study of the C. elegans nervous system and beyond
Recursive Training of 2D-3D Convolutional Networks for Neuronal Boundary Detection
Efforts to automate the reconstruction of neural circuits from 3D electron
microscopic (EM) brain images are critical for the field of connectomics. An
important computation for reconstruction is the detection of neuronal
boundaries. Images acquired by serial section EM, a leading 3D EM technique,
are highly anisotropic, with inferior quality along the third dimension. For
such images, the 2D max-pooling convolutional network has set the standard for
performance at boundary detection. Here we achieve a substantial gain in
accuracy through three innovations. Following the trend towards deeper networks
for object recognition, we use a much deeper network than previously employed
for boundary detection. Second, we incorporate 3D as well as 2D filters, to
enable computations that use 3D context. Finally, we adopt a recursively
trained architecture in which a first network generates a preliminary boundary
map that is provided as input along with the original image to a second network
that generates a final boundary map. Backpropagation training is accelerated by
ZNN, a new implementation of 3D convolutional networks that uses multicore CPU
parallelism for speed. Our hybrid 2D-3D architecture could be more generally
applicable to other types of anisotropic 3D images, including video, and our
recursive framework for any image labeling problem
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
Recommended from our members
Augmenting Wiring Diagrams of Neural Circuits with Activity in Larval Drosophila
Neural circuit models explain an animal's behavior as evoked activity of different circuit elements of its nervous system.
Synaptic wiring diagrams mapped from structural imaging of nervous systems guide modeling of neural circuits on the basis of connectivity.
However, connectivity alone may not sufficiently constrain the set of plausible circuit hypotheses for empirical study.
Combining structural imaging of synaptic connectivity with functional information from activity imaging can further constrain these hypotheses to create unequivocal neural circuit models.
This thesis develops computational methods and tools to cross-reference structural and activity imaging of explant larval Drosophila central nervous systems at cellular resolution.
Augmenting synaptic wiring diagrams with activity maps via these methods relates circuit structure and function at the neuronal level on a per-behavior basis.
Neuronal activity of larval central nervous systems expressing pan-neuronal calcium indicators is imaged in a light sheet microscope, which are then structurally imaged with high throughput electron microscopy.
Methods and tools are provided for the assembly of these image volumes, spatial registration between imaging modalities, automated detection of relevant tissue and cellular structures in each, extraction of activity time series, and morphological identification of neurons in structural imaging using reference wiring diagrams mapped from other larvae.
Using these methods, existing wiring diagrams mapped from a reference first instar larva were identified with neurons in a larva augmented with activity information for a neural circuit involved in peristaltic motor control.
This demonstrates the feasibility of the contributed methods to associate the wiring diagrams of arbitrary circuits of interest with activity timeseries across multiple individuals, behaviors, and behavioral bouts.
To demonstrate capability to augment wiring diagrams with information besides activity, these methods are also applied to multiple larvae each expressing specific neurotransmitter labels rather than calcium indicators in the light sheet microscopy.
This work scaffolds future modeling of circuits underlying behavior that can only be mechanistically understood in the context of multi-modal observation of synaptic connectivity, functional activity and molecular markers.
The methods developed also enable comparative connectomics between multiple individuals, which is necessary to study inter-individual variability in circuits and to observe experimental intervention in the development, structure, and function of neural circuits.Howard Hughes Medical Institute Janelia Research Campu
The genetic encoded toolbox for electron microscopy and connectomics
Developments in bioengineering and molecular biology have introduced a palette of genetically encoded probes for identification of specific cell populations in electron microscopy. These probes can be targeted to distinct cellular compartments, rendering them electron dense through a subsequent chemical reaction. These electron densities strongly increase the local contrast in samples prepared for electron microscopy, allowing three major advances in ultrastructural mapping of circuits: genetic identification of circuit components, targeted imaging of regions of interest and automated analysis of the tagged circuits. Together, the gains from these advances can decrease the time required for the analysis of targeted circuit motifs by over two orders of magnitude. These genetic encoded tags for electron microscopy promise to simplify the analysis of circuit motifs and become a central tool for structure‐function studies of synaptic connections in the brain. We review the current state‐of‐the‐art with an emphasis on connectomics, the quantitative analysis of neuronal structures and motifs
- …