Computed slice through an electron tomographic volume of mitochondriaisolated from mouse liver and treated with the serine protease inhibitor MG132.
Full resolution image description
Zip file (RudyD_vol.zip) containing the tomographic reconstruction in Analyze 7.5 format: RudyD_sub.img/hdr. This file contains the full sized reconstruction, despite its name. A subvolume of this was used for segmentation (see image map file). This subvolume is available with the segmentation file download.
2017, 3006, 271
0.0014, 0.0014, 0.0014
Animation through a portion of the tomographic volume of isolated mitochondria treated with the serine protease inhibitor MG132. The animation was downsampled for ease of viewing. (RudyD_vol.avi)
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 (RudyD.trace) containing the manual traces and the subvolume in Analyze 7.5 format (RudyD_sub1.img/.hdr files) used for tracing. Surfaces generated from the contours for each object are included in Synu format (*.synu). Note that both Xvoxtrace and synu can now be read using Jinx, available from the CCDB download page.
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 with 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:4096, mus musculus, mitochondrion. CIL. Dataset. https://doi.org/doi:10.7295/W9CCDB4096