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- EMDB-15443: In situ cryo-electron tomogram of a Taxol treated intact pluripot... -

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Basic information

Entry
Database: EMDB / ID: EMD-15443
TitleIn situ cryo-electron tomogram of a Taxol treated intact pluripotent P19 cell.
Map dataIn situ cryo-electron tomogram of a Taxol treated intact pluripotent P19 cell.
Sample
  • Cell: In situ cryo-electron tomogram of a Taxol treated intact pluripotent P19 cell.
KeywordsMicrotubules / Scaffold / protein transport
Biological speciesMus musculus (house mouse)
Methodelectron tomography / cryo EM
AuthorsChakraborty S / Martinez-Sanchez A / Baumeister W / Mahamid J
Funding support Germany, 1 items
OrganizationGrant numberCountry
Other government Germany
CitationJournal: To Be Published
Title: In situ cryo-electron tomogram of an intact human induced pluripotent stem cell derived neuronal process
Authors: Chakraborty S / Martinez-Sanchez A / Baumeister W / Mahamid J
History
DepositionJul 21, 2022-
Header (metadata) releaseJan 31, 2024-
Map releaseJan 31, 2024-
UpdateJan 31, 2024-
Current statusJan 31, 2024Processing site: PDBe / Status: Released

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Structure visualization

Supplemental images

emd_15443.png

Downloads & links

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Map

FileDownload / File: emd_15443.map.gz / Format: CCP4 / Size: 102.7 MB / Type: IMAGE STORED AS SIGNED INTEGER (2 BYTES)
AnnotationIn situ cryo-electron tomogram of a Taxol treated intact pluripotent P19 cell.
Voxel sizeX=Y=Z: 27.36 Å
Density
Minimum - Maximum-96.0 - 75.0
Average (Standard dev.)2.6191928 (±5.3814154)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin00-125
Dimensions464464250
Spacing464464250
CellA: 12695.04 Å / B: 12695.04 Å / C: 6840.0 Å
α=β=γ: 90.0 °

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Supplemental data

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Sample components

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Entire : In situ cryo-electron tomogram of a Taxol treated intact pluripot...

EntireName: In situ cryo-electron tomogram of a Taxol treated intact pluripotent P19 cell.
Components
  • Cell: In situ cryo-electron tomogram of a Taxol treated intact pluripotent P19 cell.

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Supramolecule #1: In situ cryo-electron tomogram of a Taxol treated intact pluripot...

SupramoleculeName: In situ cryo-electron tomogram of a Taxol treated intact pluripotent P19 cell.
type: cell / ID: 1 / Parent: 0
Details: The cells were treated with Taxol for 30 minutes prior to vitrification
Source (natural)Organism: Mus musculus (house mouse)

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Experimental details

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Structure determination

Methodcryo EM
Processingelectron tomography
Aggregation statecell

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Sample preparation

BufferpH: 7.2
GridModel: Quantifoil R2/1 / Material: GOLD / Mesh: 200 / Pretreatment - Type: GLOW DISCHARGE / Pretreatment - Time: 45 sec.
VitrificationCryogen name: ETHANE-PROPANE / Chamber humidity: 95 % / Chamber temperature: 310 K / Instrument: FEI VITROBOT MARK IV
Details: blot force 10, blot time 10 seconds from backside..
DetailsCells are cultured on EM grid
SectioningOther: NO SECTIONING

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Electron microscopy

MicroscopeTFS KRIOS
Electron beamAcceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron opticsC2 aperture diameter: 70.0 µm / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELDBright-field microscopy / Cs: 2.7 mm / Nominal defocus max: 0.5 µm / Nominal defocus min: 0.2 µm / Nominal magnification: 42000
Specialist opticsPhase plate: VOLTA PHASE PLATE / Energy filter - Name: GIF Quantum LS / Energy filter - Slit width: 20 eV
Sample stageSpecimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN
TemperatureMin: 77.0 K
Image recordingFilm or detector model: GATAN K2 SUMMIT (4k x 4k) / Detector mode: COUNTING / Average electron dose: 1.4 e/Å2
Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company

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Image processing

Final reconstructionAlgorithm: BACK PROJECTION / Software - Name: IMOD (ver. 4.9.0) / Number images used: 61

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