ELECTRON MICROSCOPY

Sample

Closed conformation dome complex including the Sec-translocon (SecYEG-SecA-SecDF) in untreated Mycoplasma pneumoniae cells

Specimen Preperation
Sample Aggregation State CELL
Vitrification Instrument HOMEMADE PLUNGER
Cryogen Name ETHANE-PROPANE
Sample Vitrification Details Back-side blotting for 2-3 seconds before plunging using a manual plunger without an environmental chamber.
3D Reconstruction
Reconstruction Method SUBTOMOGRAM AVERAGING
Number of Particles 11557
Reported Resolution (Å) 9.1
Resolution Method FSC 0.143 CUT-OFF
Other Details ?
Refinement Type
Symmetry Type POINT
Map-Model Fitting and Refinement
ID 1
Refinement Space REAL
Refinement Protocol RIGID BODY FIT
Refinement Target ?
Overall B Value ?
Fitting Procedure ?
Details Fitting was performed using PowerFit and Fit-in-map in ChimeraX
Data Acquisition
Detector Type GATAN K2 SUMMIT (4k x 4k)
Electron Dose (electrons/Å2) 2.9
Imaging Experiment
Date of Experiment ?
Temprature (Kelvin)
Microscope Model TFS KRIOS
Minimum Defocus (nm) 1500
Maximum Defocus (nm) 3500
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 81000
Calibrated Magnification ?
Source FIELD EMISSION GUN
Acceleration Voltage (kV) 300
Imaging Details ?
Imaging Experiment
Task Software Package Version
VOLUME SELECTION Warp 2.0.0
IMAGE ACQUISITION SerialEM ?
CTF CORRECTION Warp 2.0.0
MODEL FITTING PowerFit ?
MODEL FITTING UCSF ChimeraX ?
FINAL EULER ASSIGNMENT Warp 2.0.0
CLASSIFICATION RELION 4.0.1
RECONSTRUCTION Warp 2.0.0
Image Processing
CTF Correction Type CTF Correction Details Number of Particles Selected Particle Selection Details
PHASE FLIPPING AND AMPLITUDE CORRECTION ?
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