ELECTRON MICROSCOPY

Sample

Saccharomyces cerevisiae large 60S ribosomal subunit

Specimen Preperation
Sample Aggregation State PARTICLE
Vitrification Instrument HOMEMADE PLUNGER
Cryogen Name ETHANE
Sample Vitrification Details A 3 uL sample was applied onto a freshly plasma-treated (6 seconds, Gatan Solarus plasma cleaner) holey carbon C-flat grid (Protochips, Inc.), allowing the sample to adsorb for 30 seconds. The sample was then plunge-frozen in liquid ethane using a manual cryo-plunger in ambient atmosphere at 4 degrees C.
3D Reconstruction
Reconstruction Method SINGLE PARTICLE
Number of Particles 75653
Reported Resolution (Å) 2.9
Resolution Method FSC 0.143 CUT-OFF
Other Details B-factor of -50 applied to final map (Single particle--Applied symmetry: C1).
Refinement Type
Symmetry Type POINT
Map-Model Fitting and Refinement
ID 1
Refinement Space REAL
Refinement Protocol FLEXIBLE FIT
Refinement Target FSC
Overall B Value 50
Fitting Procedure ?
Details REFINEMENT PROTOCOL--flexible
Data Acquisition
Detector Type GATAN K2 (4k x 4k)
Electron Dose (electrons/Å2) 25
Imaging Experiment
Date of Experiment 2014-09-02
Temprature (Kelvin)
Microscope Model FEI TITAN KRIOS
Minimum Defocus (nm) 500
Maximum Defocus (nm) 2500
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 22500
Calibrated Magnification 38167
Source FIELD EMISSION GUN
Acceleration Voltage (kV) 300
Imaging Details super-resolution imaging
Imaging Experiment
Task Software Package Version
MODEL FITTING Rosetta ?
MODEL FITTING UCSF Chimera ?
RECONSTRUCTION FREALIGN ?
Image Processing
CTF Correction Type CTF Correction Details Number of Particles Selected Particle Selection Details
. each particle
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