EMD-5375 Experiments and Validation

Direct electron detection yields cryo-EM reconstructions at resolutions beyond .75 Nyquist frequency

Single particle reconstruction
Overview of EMD-5375
Sample name: Bacteriophage P22 procapsid
Organism: Enterobacteria phage P22
Related EM entry by publication: EMD-5377

Map parameters

Recommended contour level: 0.80 (author)
Number of grid points: 384 × 384 × 384
Voxel size: 3.48 × 3.48 × 3.48 Å
Minimum density: -0.999
Maximum density: 1.869
Average density: 0.00
Standard deviation: 0.181

Sample information

Sample name: Bacteriophage P22 procapsid
Virus: Enterobacteria phage P22

Validation information

Experimental information

 

Image processing

Applied symmetry: I
Software: MPSA
Number of particles: 7500
Reconstruction protocol: Cross-common Lines
CTF correction: Each Micrograph
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.

Imaging

Session
Microscope model: JEOL 2010F
Electron source: FIELD EMISSION GUN
Electron dose (e/A**2): 17.0
Nominal CS (mm): 2.00
Nominal magnification: 15000.0
Defocus max (nm): 5,000.00
Defocus min (nm): 2,000.00
Holder model: GATAN LIQUID NITROGEN
Acquisition
Number of images: 200
Sampling interval (μm): 6.0
Details: Images were collected on a CMOS type detector DE-12 from Direct Electron

Specimen

Preparation
Specimen state: particle
Specimen concentration (mg/mL): 1.00
Buffer: 50 mM Tris pH 7.6, 25 mM NaCl, 2mM EDTA
Specimen support: 400 mesh copper grid
Vitrification
Apparatus: FEI VITROBOT
Cryogen: ETHANE
Humidity: 95
Protocol: Blot for 2 seconds before plunging
Details: Vitrification instrument: Vitrobot