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Licensing
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*CIL – Cell Image Library accession number. Please use this to reference an image.

CIL:40178*  Cite 
Description

Single computed slice through a dual axis tilt electron tomographic reconstruction of double membrane vesicles in a freeze-substituted VeroE6 cultured cell infected with the SARS-CoV. This image has been downsampled from the raw data image which can be accessed using the link provided to the Cell Centered Database. For more information, see: Knoops et al. SARS-coronavirus replication is supported by a reticulovesicular network of modified endoplasmic reticulum. PLoS Biol. 2008 6(9):e226. PMID: 18798692

Technical Details

For ultrastructural morphological investigations, SARS-CoV-infected Vero E6 cells were pre-fixed (for biosafety reasons) overnight with 3% paraformaldehyde in 0.1M PHEM buffer (60 mM piperazide-1,4-bis[2-ethanesulfonic acid], 25 mM HEPES, 2 mM MgCl2, 10 mM EGTA) at various time points after infection. For cryo-fixation, cell monolayers adhered to Thermanox coverslips (Nunc, Denmark) were plunged into liquid ethane. Freeze substitution was performed at -90°C in an automated freeze-substitution system (Leica, Austria) using an FS medium consisting of 90% acetone and 10% water, containing 1% osmium tetroxide and 0.5 % uranyl acetate. After washing with pure acetone at RT, the samples were embedded in epoxy LX-12 resin. Thin sections were collected on 100 Mesh grids, 1.25% pioloform support film contrasted with uranyl acetate and lead hydroxide and subsequently viewed at 80 kV with a Philips CM-10 transmission electron microscope (Philips, The Netherlands).

Biological Sources
NCBI Organism Classification
Chlorocebus pygerythrus
Cell Type
epithelial cell
Cell Line
Vero
Cellular Component
plasma membrane
Biological Context
Biological Process
response to virus
Attribution
Names
Kèvin Knoops
Abraham J. Koster
A. Mieke Mommaas
Eric J. Snijder
Link
CCDB 6021
Citation
Digital Object Identifier (DOI)
doi:10.7295/W9CIL40178
Archival Resource Key (ARK)
ark:/b7295/w9cil40178
Grouping This image is part of a group.
Imaging
Image Type
recorded image
Image Mode
transmission electron microscopy (TEM)
Parameters Imaged
electron density
Visualization Methods
uranyl salt
Data Qualifiers
processed data
Sample Preparation
Methods
osmium tetroxide fixed tissue
FBbi:00000621
Relation To Intact Cell
dispersed cells in vitro
Dimensions
Spatial Axis Image Size Pixel Size
X 512px ——
X 512px ——