Single tilt image at zero degrees tilt from a tilt series taken through +/- 60 degrees using an intermediate voltage electron microscope of mitochondria isolated from mouse liver treated with both the serine protease inhibitor MG132 and BH3 peptide from Bid.
Full resolution image description
Zip file containing the tilt images after pre alignment using cross correlation in IMOD (RudyH.preali); the original digitized unaligned images were not available. Also included are the other supporting files generated by IMOD. Some may also be used by TxBr.
Manual segmentation of the individual cristae and inner and outer mitochondrial membranes using Xvoxtrace 2.18. Contours were traced every other section on even numbered planes.
Segmentation file description
Zip file containing the .trace file generated by Xvoxtrace for the manual contours; and the surface objects generaged by Synu (*.synu). Also included is the Viewdata file required to view the surfaced objects using Synuview. Note that these files may now by opened and viewed using Jinx (see CCDB tools). To view the manual traces superimposed on the volume, the reconstruction file must also be downloaded.
Proapoptotic BH3-only proteins induce Bax/Bak-dependent mitochondrial cristae remodeling independent of cytochrome c release and Bak oligomerization
Don Newmeyer; Lydia Lartigue; Guy Perkins
Ph.D.; Ray T Scott; Amruta Dixit;
Cytochrome C Release Assay
To determine the effect of pro-apoptotic peptides on mitochondrial morphology and cytochrome C release.
Microscopy product ID
Mitochondria were incubated Bid-BH3 peptides and the serine protease inhibitor MG132 at 37 degrees C for 12 min in AT buffer containing 10 mM KCl and centrifuged at 5200x g for 6 min.
Tissue product storage
Electron microscopy product
Mouse liver mitochondria were prepared as described (Yamaguchi et al., 2006).Pelleted mitochondria were fixed with a 37 C solution of 2% paraformaldehyde, 2.5% glutaraldehyde (Ted Pella) in 0.15 M sodium cacodylate (pH 7.4), and then incubated for an additional 30 minutes on ice. Fixed samples were then rinsed 3 times for 3 minutes each with 0.15 M sodium cacodylate plus 3 mM calcium chloride (pH 7.4) on ice, post-fixed with 1 percent osmium tetroxide, 0.8% potassium ferrocyanide, 3 mM calcium chloride in 0.15 M sodium cacodylate (pH 7.4) for 60 minutes, and then washed 3 times for 3 minutes with ice-cold distilled water. The samples were stained overnight with 2% uranyl acetate at 4 degrees C, dehydrated in graded ethanol baths, and embedded in Durcupan ACM resin (Fluka).Sections from the embedded mitochondria samples were cut at thicknesses of nominally 500 nm. Sections were then stained 30 min in 2 percent aqueous uranyl acetate, followed by 15 min in lead salts. Fiducial cues consisting of 15 and 20 nm colloidal gold particles were deposited on opposite sides of the section
Imaging product type
tilted using a computer controlled goniometer about an axis perpendicular to
the optical axis of the microscope
Ryuji Yamaguchi, Ph.D., Don Newmeyer; Lydia Lartigue; Guy Perkins, Ph.D.; Ray T Scott; Amruta Dixit; CCDB:4098, mus musculus, mitochondrion. CIL. Dataset. https://doi.org/doi:10.7295/W9CCDB4098