Alternate header for print version


Reconstruction
Display image description
Computed slice through an electron tomographic volume of mitochondria isolated from mouse liver and treated with both the protease inhibitor MG132 and BH3 peptide from Bid.
Full resolution image description
Zip file containing the reconstruction file in Analyze 7.5 format: RudyH_newsub.img/.hdr. This reconstruction was used to generate the segmentation shown in the "Segmentation" window. (RudyH_vol.zip)
Volume_dimension
1400, 2000, 324
Volume scale
0.0014, 0.0014, 0.0014
Animation description
Animation through the computed slices of an electron tomographic volume of mitochondria isolated from mouse liver and treated with both the protease inhibitor MG132 and BH3 peptide from Bid.

Image 2D
Display image description
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.

Segmentation
Display image description
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.

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

CCDB:4098*  Cite 
Project: P1714
Project name
Remodeling of Crista During Cytochrome C Release
Description
Proapoptotic BH3-only proteins induce Bax/Bak-dependent mitochondrial cristae remodeling independent of cytochrome c release and Bak oligomerization
Leader(s)
Ryuji Yamaguchi
Ph.D.
Collaborator(s)
Don Newmeyer; Lydia Lartigue; Guy Perkins
Ph.D.; Ray T Scott; Amruta Dixit;
Start date
unspecified
End date
unspecified
 
Experiment
Experiment ID
3458
Title
Cytochrome C Release Assay
Purpose
To determine the effect of pro-apoptotic peptides on mitochondrial morphology and cytochrome C release.
Experimenter(s)
Guy Perkins
Microscopy product
Microscopy product ID
4098
Instrument
JEOL4000EX IVEM
Microscopy type
IVEM
Product type
SINGLE TILT
Image basename
Rudy H
Subject
Species
mouse
Scientific name
mus musculus
Strain
unknown
Group by
Drug treatment
Treatment
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.
Age class
adult
Tissue section
Anatomical location
liver
Tissue product storage
liquid nitrogen
Thickness
0.5 µm
Specimen description
Organ
liver
Structure
mitochondrion
Imaging parameters
Type
Electron microscopy product
Recording medium
film
Magification
20000
Accelerating voltage
400 kV
Specimen preparation
Protocol used
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
Type
Single tilt
Description
singlet_desc
Min range
-60 degrees
Max range
60 degrees
Tilt increment
2 degrees
Notes
tilted using a computer controlled goniometer about an axis perpendicular to the optical axis of the microscope