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Reconstruction
Display image description
Computed slice through a tomographic reconstruction of the pre- and post-synaptic structures of the calycal synapse onto a chick ciliary ganglion neuron. This slice was derived from the 3rd of 6 serial volumes comprising the entire reconstruction.
Full resolution image description
Zip file containing entire serial tomographic volume in Analyze 7.5 format (embryo.hdr/embryo.img). This volume was creating by aligning and concatenating 6 serial tomograms. The volume file is 269.5 Mb.
Volume_dimension
640, 640, 690
Animation description
Downsampled animation (256x256) through the computed slices of the aligned and merged volumes showing the pre- and post-synaptic structures of the calycal synapse onto a chick ciliary ganlgion neuron.

Segmentation
Display image description
Manual delineation of the presynaptic calyx and post synaptic spine mat using Xvoxtrace 2.17. Also delineated were the locations of PSD's, vesicles, intracellular components of the post synaptic compartment.
Segmentation file description
Zip file (~38 Mb) containing the Xvoxtrace files (*.trace) containing the manual tracings of the individual components of the calycyl synapse, including ER, PSD, vesicles, base, and spines. Each of these components was traced in a separate file and are contained in different subdirectories. Also included in each subdirectory are the surfaced contours in Synu (*.synu) format, along with the Viewdata file required for visualization with Synuview.

License
Attribution Only: This image is licensed under a Creative Commons Attribution License. View License Deed | View Legal Code

CCDB:3629*
Project: P1692
Project name
Reconstruction of ciliary ganglion spine mats in adult and embryonic chick
Description
Detailed structural investigation of the structure and distribution of synaptic densities on the spine mats from adult and embryonic ciliary ganglion neurons. Reconstructions were derived from serial electron tomography of in vivo neurons.
Funding agency
NIH
Leader(s)
Rick Shoop
Collaborator(s)
Darwin Berg
Mark Ellisman
Naoko Yamada
Ed Esquenazi
Start date
unspecified
End date
unspecified
 
Experiment
Experiment ID
3422
Title
Serial tomography of embryonic ciliary ganglion spine mat
Purpose
To employ serial electron tomography to reconstruct an entire spine mat from embryonic chick ciliary ganglion.
Experimenter(s)
Rick Shoop
Microscopy product
Microscopy product ID
3629
Instrument
JEOL 4000EX IVEM
Microscopy type
IVEM
Product type
SINGLE TILT
Image basename
embryo
Subject
Species
chicken
Scientific name
Gallus gallus
Strain
White leghorn
Group by
age
Age
15 days
Age class
embryonic
Tissue section
Anatomical location
ciliary ganglion
Microtome
ultramicrotome
Thickness
100 µm
Specimen description
Organ
ciliary ganglion
System
peripheral nervous system
Structure
calycal synapse
Cell type
ciliary ganglion neuron
Imaging parameters
Type
Electron microscopy product
Recording medium
film
Magification
10000
Accelerating voltage
400 KeV
Specimen preparation
Protocol used
Embryonic day (E) 15 chicks or 2-year-old adult chickens were perfusion-fixed with 2% paraformaldehyde plus 2% glutaraldehyde in cacodylate buffer, pH 7.4. Ciliary ganglia were removed, transferred to fresh fixative, and incubated for 3 hr at room temperature. Adult ciliary ganglia were cut into 1 mm3 pieces. After being rinsed several times in 0.1 M sodium cacodylate buffer, pH 7.4, the tissue was treated for 30 min with 2% osmium tetroxide in 0.1 M sodium cacodylate and then counterstained with uranyl acetate. The ganglia were dehydrated in a series of ethanol solutions followed by two rinses in acetone, infiltrated with Durcupan ACM resin (Electron Microscopy Sciences, Fort Washington, PA), allowed to polymerize for 24 hr at 60C, and then sectioned. For serial tomography a continuous series of 10 1-m-thick sections was prepared to encompass a complete spine mat. For traditional thin section analysis 100-nm-thick sections were made.
Imaging product type
Type
Single tilt
Description
singlet_desc
Min range
-60 degrees
Max range
60 degrees
Tilt increment
2 degrees
Notes
The reconstruction shown is actually from a serial tomogram derived from 6 serial sections. As far as can be derived from the published paper, all tilt series were -60 to 60 at 2 degree increments.