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- EMDB-51759: Dual-Axis cryo-STEM tomogram of a Mitochondrion from MFF-/- MEFs -
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Open data
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Basic information
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Title | Dual-Axis cryo-STEM tomogram of a Mitochondrion from MFF-/- MEFs | |||||||||||||||
![]() | Dual-axis BF-STEM cryo-Tomogram of MFF-/- MEFs | |||||||||||||||
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![]() | Mitochondrial morphology / whole cell tomography / cryo-STEM Tomography / CSTET / UNKNOWN FUNCTION | |||||||||||||||
Biological species | ![]() ![]() | |||||||||||||||
Method | electron tomography / cryo EM | |||||||||||||||
![]() | Kirchweger P / Wolf SG / Elbaum M / Fass D | |||||||||||||||
Funding support | ![]() ![]()
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![]() | ![]() Title: Snapshots of mitochondrial fission imaged by cryo-scanning transmission electron tomography. Authors: Peter Kirchweger / Sharon Grayer Wolf / Neta Varsano / Tali Dadosh / Guenter P Resch / Michael Elbaum / ![]() ![]() Abstract: Mitochondria undergo constant remodeling via fission, fusion, extension and degradation. Fission, in particular, depends on the accumulation of mitochondrial fission factor (MFF) and subsequent ...Mitochondria undergo constant remodeling via fission, fusion, extension and degradation. Fission, in particular, depends on the accumulation of mitochondrial fission factor (MFF) and subsequent recruitment of the dynamin-related protein DRP1 (also known as DNM1L). We used cryo-scanning transmission electron tomography (cryo-STET) to investigate mitochondrial morphologies in MFF mutant (MFF-/-) mouse embryonic fibroblast (MEF) cells in ATP-depleting conditions that normally induce fission. The capability of cryo-STET to image through the cytoplasmic volume to a depth of 1 µm facilitated visualization of intact mitochondria and their surroundings. We imaged changes in mitochondrial morphology and cristae structure, as well as contacts with the endoplasmic reticulum (ER), degradative organelles and the cytoskeleton at stalled fission sites. We found disruption of the outer mitochondrial membrane at contact sites with the ER and degradative organelles at sites of mitophagy. We identified fission sites where the inner mitochondrial membrane is already separated while the outer membrane is still continuous. Although MFF is a general fission factor, these observations demonstrate that mitochondrial fission can proceed to the final stage in its absence. The use of cryo-STET allays concerns about the loss of structures due to sample thinning required for tomography using cryo-transmission electron microscopy. #1: ![]() Title: Snapshots of Mitochondrial Fission Imaged by Cryo-Scanning Transmission Electron Tomography Authors: Kirchweger P / Wolf SG / Varsano N / Dadosh T / Resch GP / Elbaum M | |||||||||||||||
History |
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Structure visualization
Supplemental images |
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Downloads & links
-EMDB archive
Map data | ![]() | 4.7 GB | ![]() | |
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Header (meta data) | ![]() ![]() | 11.9 KB 11.9 KB | Display Display | ![]() |
Images | ![]() | 188.5 KB | ||
Filedesc metadata | ![]() | 4.3 KB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Validation report
Summary document | ![]() | 322.7 KB | Display | ![]() |
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Full document | ![]() | 322.3 KB | Display | |
Data in XML | ![]() | 4 KB | Display | |
Data in CIF | ![]() | 4.6 KB | Display | |
Arichive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
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Links
EMDB pages | ![]() ![]() |
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Map
File | ![]() | ||||||||||||||||||||||||||||||||
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Annotation | Dual-axis BF-STEM cryo-Tomogram of MFF-/- MEFs | ||||||||||||||||||||||||||||||||
Projections & slices | Image control
Images are generated by Spider. generated in cubic-lattice coordinate | ||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 20.42 Å | ||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||
Details | EMDB XML:
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-Supplemental data
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Sample components
-Entire : Mouse embryonic fibroblast
Entire | Name: Mouse embryonic fibroblast |
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Components |
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-Supramolecule #1: Mouse embryonic fibroblast
Supramolecule | Name: Mouse embryonic fibroblast / type: cell / ID: 1 / Parent: 0 Details: Mitochondrial fission factor deletion MEF, expressing a mito-GFP |
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Source (natural) | Organism: ![]() ![]() |
-Experimental details
-Structure determination
Method | cryo EM |
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![]() | electron tomography |
Aggregation state | cell |
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Sample preparation
Buffer | pH: 7.4 / Details: DMEM growth media |
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Grid | Model: Quantifoil R2/2 / Material: GOLD / Mesh: 200 / Support film - Material: CARBON / Support film - topology: HOLEY / Pretreatment - Type: GLOW DISCHARGE / Pretreatment - Time: 120 sec. / Pretreatment - Atmosphere: AIR |
Vitrification | Cryogen name: ETHANE / Chamber humidity: 95 % / Chamber temperature: 310.15 K / Instrument: LEICA EM GP |
Sectioning | Other: NO SECTIONING |
Fiducial marker | Manufacturer: Home made / Diameter: 15 nm |
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Electron microscopy
Microscope | TFS KRIOS |
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Image recording | Film or detector model: OTHER / Digitization - Dimensions - Width: 4096 pixel / Digitization - Dimensions - Height: 4096 pixel / Average electron dose: 1.05 e/Å2 Details: Images were recorded on a Fischione HAADF detector in BrightField mode (described in Kirchweger et al., 2023). |
Electron beam | Acceleration voltage: 300 kV / Electron source: ![]() |
Electron optics | C2 aperture diameter: 70.0 µm / Calibrated defocus min: 0.0 µm / Illumination mode: SPOT SCAN / Imaging mode: BRIGHT FIELD / Nominal defocus max: 0.0 µm / Nominal defocus min: 0.0 µm / Nominal magnification: 41000 |
Sample stage | Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN |
Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |
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Image processing
Final reconstruction | Algorithm: BACK PROJECTION Software:
Details: After WBP, the tomogram was deconvolved as described in Waugh et al. (2021) and Kirchweger et al. (2023) Number images used: 122 | ||||||
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CTF correction | Details: CSTET needs no CTF correction / Type: NONE |