ELECTRON MICROSCOPY

Sample

Pentamer of the actinorhodopsin

Specimen Preperation
Sample Aggregation State PARTICLE
Vitrification Instrument FEI VITROBOT MARK IV
Cryogen Name ETHANE
Sample Vitrification Details In brief, grids were incubated for 30 seconds, and excess liquid blotted off for 5 seconds applying a blotting force of -6 at 4 degrees and a relative humidity of about 100% before plunging into liquid ethane (-177 degrees to -171 degrees).
3D Reconstruction
Reconstruction Method SINGLE PARTICLE
Number of Particles 184961
Reported Resolution (Å) 2.84
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 Shiff base bond created between K234 and RETINAL (RET)
Data Acquisition
Detector Type TFS FALCON 4i (4k x 4k)
Electron Dose (electrons/Å2) 1.01
Imaging Experiment
Date of Experiment ?
Temprature (Kelvin)
Microscope Model TFS KRIOS
Minimum Defocus (nm) 1000
Maximum Defocus (nm) 2200
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 165000
Calibrated Magnification ?
Source FIELD EMISSION GUN
Acceleration Voltage (kV) 300
Imaging Details ?
Imaging Experiment
Task Software Package Version
PARTICLE SELECTION crYOLO 1.5
IMAGE ACQUISITION SerialEM ?
CTF CORRECTION Gctf 1.6
MODEL FITTING UCSF Chimera 1.7.1
INITIAL EULER ASSIGNMENT RELION 4
FINAL EULER ASSIGNMENT cryoSPARC ?
CLASSIFICATION RELION 4
RECONSTRUCTION cryoSPARC ?
MODEL REFINEMENT PHENIX 1.20-4459
Image Processing
CTF Correction Type CTF Correction Details Number of Particles Selected Particle Selection Details
PHASE FLIPPING AND AMPLITUDE CORRECTION ?
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