Single tilt image of a 250 um thick section of a Drosophila S2 cell infected with flock house virus (FHV-1) imaged with intermediate voltage electron microscopy, taken at zero degree tilt. The dark specks are colloidal gold particles that have been applied to the surfaces of the section to serve as fiducial marks for subsequent alignment. 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: Kopek et al. Three-dimensional analysis of a viral RNA replication complex reveals a virus-induced mini-organelle. PLoS Biol. 2007 Sep;5(9):e220. PMID: 17696647
Mitochondria were isolated from Drosophila cells as described by Echalier . For electron tomography, cells were fixed in 2% parafomaldehyde and 2.5% glutaraldehyde in 0.1 M sodium cacodylate, pH 7.4, post-fixed in 1% osmium tetroxide with 0.8% potassium ferrocyanide in sodium cacodylate buffer, stained with 2% uranyl acetate, dehydrated in a graded series of ethanol, and embedded in Durcupan ACM resin. Sections were cut with a thickness of approximately 250 nm from blocks exhibiting well-preserved ultrastructure. These sections were stained for 30 min in 2% aqueous uranyl acetate, followed by 30 min in lead salts. Fiducial cues consisting of 20-nm colloidal gold particles were deposited on both sides of the section.
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