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Title:Direct electron detection yields cryo-EM reconstructions at resolutions beyond .75 Nyquist frequency
Authors:Bammes BE, Rochat RH, Jakana J, Chen D, Chiu W
Sample:Bacteriophage P22 procapsid
Method:Single particle reconstruction (8.5 angstroms resolution)
Sample
Sample name: Bacteriophage P22 procapsid
Components:
ID Type Name Exp. MW (MDa) Theo. MW (MDa) Oligomeric details Recombinant expression Synthetic Organism UniProt identifier GO identifier InterPro identifier Virus identifier Details
1virusEnterobacteria phage P22Enterobacteria phage P22
Experiment
Specimen state: Particle
Specimen preparation:
pHSpecimen conc.DetailsStainingSpecimen support details
7.61 mg/mL50 mM Tris pH 7.6, 25 mM NaCl, 2mM EDTA400 mesh copper grid
Vitrification:
Cryogen nameHumidityTemp.Instr.MethodTime resolvedDetails
ETHANE95%170 KFEI VITROBOTBlot for 2 seconds before plunging msVitrification instrument: Vitrobot
Imaging:
MicroscopeVoltageIllumination modeImaging modeCsDefocus min.Defocus max.Nominal mag.Calibrated mag.Electron sourceDetectorDetector distanceAstigmatism
JEOL 2010F200 kVFLOOD BEAMBRIGHT FIELD2.0 mm2000 nm5000 nm1500017200FIELD EMISSION GUNDIRECT ELECTRON DE-12 (4k x 3k) mmobjective lens astigmatism was corrected at 100,000 times magnification

Specimen holderHolder modelTilt min.Tilt max.Energy filterEnergy windowTemp.Temp. min.Temp. max.Beam tiltElectron doseOther detailsDate
Side EntryGATAN LIQUID NITROGEN°° eV100 K100 K100 K mrad17 e/Å210-JUN-2010
Processing
Protocol:Cross-common Lines
Software:MPSA
CTF correction:Each Micrograph
Number of particles:7500
Imposed symmetry:I
Resolution by author:8.5 Å
Resolution method:FSC 0.5
Processing details:Resolution was calculated to be 8.5 Angstroms at 0.5 FSC and 7.3 Angstroms at 0.143 FSC as compared against the 3.8 Angstrom reconstruction of P22.
Scanned images:
Num. imagesSampling sizeOD rangeQuant. bit numberOther detailsScanner
2006 μm/pixelImages were collected on a CMOS type detector DE-12 from Direct Electron
Fitting:
PDBProtocolTarget crit.SoftwareB valueFitting spacePDB chainDetails