ELECTRON MICROSCOPY

Sample

Neuro2a expressing Q150-GFP aggregate

Specimen Preperation
Sample Aggregation State PARTICLE
Vitrification Instrument FEI VITROBOT MARK IV
Cryogen Name ETHANE-PROPANE
Sample Vitrification Details ?
3D Reconstruction
Reconstruction Method SINGLE PARTICLE
Number of Particles 43227
Reported Resolution (Å) 3.9
Resolution Method FSC 0.143 CUT-OFF
Other Details ?
Refinement Type
Symmetry Type POINT
Map-Model Fitting and Refinement
ID 1
Refinement Space ?
Refinement Protocol ?
Refinement Target ?
Overall B Value ?
Fitting Procedure ?
Details ?
Data Acquisition
Detector Type FEI FALCON IV (4k x 4k)
Electron Dose (electrons/Å2) 2
Imaging Experiment
Date of Experiment ?
Temprature (Kelvin)
Microscope Model TFS KRIOS
Minimum Defocus (nm) 3000
Maximum Defocus (nm) 5000
Minimum Tilt Angle (degrees) ?
Maximum Tilt Angle (degrees) ?
Nominal CS 2.7
Imaging Mode BRIGHT FIELD
Specimen Holder Model FEI TITAN KRIOS AUTOGRID HOLDER
Nominal Magnification ?
Calibrated Magnification ?
Source FIELD EMISSION GUN
Acceleration Voltage (kV) 300
Imaging Details For dataset 2, 410 tilt series were collected on a Titan Krios G4 instrument at 300 kV equipped with a Selectris X energy filter operating at zero-loss with a slit width of 10 eV, and a Falcon 4i camera (Thermo). Low-magnification images were taken at 11500x (object pixel size 2.13 nm) to generate lamella overviews. Tilt series were recorded at 1.89 A pixel size using the Tomography 5 (Thermo) in EER file format, with a dose-symmetric tilt scheme. The tilt series were collected with a 3 degree tilt increment with an angular range of ~120 degrees, and with cumulative dose of 80-120 electrons/A^2. Targeted defocus was in the range of -5 to -3 um.
Imaging Experiment
Task Software Package Version
IMAGE ACQUISITION FEI tomography 9.03
MODEL REFINEMENT PHENIX ?
RECONSTRUCTION RELION 4.0
Image Processing
CTF Correction Type CTF Correction Details Number of Particles Selected Particle Selection Details
NONE ?
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