Instrument: HOMEMADE PLUNGER / Cryogen name: ETHANE / Temperature: 295 K / Humidity: 100 % Details: The assembled sample (1.5 microliter) was applied to the carbon side of a glow discharged perforated Quantifoil grid, followed by application of 3 microliter of low salt buffer (100 milimolar NaCl, 50 milimolar Tris pH 8.0) on the back side of the grid, and blotting, from the back side, with a filter paper, before plunge-freezing in liquid ethane.
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Electron microscopy imaging
Experimental equipment
Model: Tecnai Polara / Image courtesy: FEI Company
Imaging
Microscope: FEI POLARA 300
Electron gun
Electron source: FIELD EMISSION GUN / Accelerating voltage: 300 kV / Electron dose: 41 e/Å2 / Illumination mode: FLOOD BEAM
Lens
Magnification: 31000.0 X (nominal) / Cs: 2.2 mm / Imaging mode: BRIGHT FIELD / Defocus: 700.0 - 2200.0 nm
Specimen Holder
Model: SIDE ENTRY, EUCENTRIC
Camera
Detector: GATAN K2 SUMMIT (4k x 4k)
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Image acquisition
Image acquisition
Number of digital images: 523
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Image processing
Processing
Method: helical reconstruction
3D reconstruction
Algorithm: FOURIER SPACE / Software: RELION / Resolution: 5 Å / Resolution method: FSC 0.143 CUT-OFF
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