3J0B
cryo-EM reconstruction of West Nile virus
EM specimen
| Vitrification | Yes |
| Instrument | HOMEMADE PLUNGER |
| Method | A small vial of ethane is placed inside a larger liquid nitrogen reservoir. The grid holding a few microliters of the sample is held in place at the bottom of a plunger by means of fine tweezers. Once the ethane in the vial is completely frozen, it needs to be melted slightly. When the liquid ethane is ready, a piece of filter paper is then pressed against the sample to blot off excess buffer sufficient to leave a thin layer on the grid. After a predetermined time, the filter paper is removed and the plunger is allowed to drop into the liquid ethane. Once the grid enters the liquid ethane, the sample is rapidly frozen and the grid is transferred under liquid nitrogen to a storage box immersed in liquid nitrogen for later use in the microscope. |
EM Data Collection
| Microscope | FEI/PHILIPS CM300FEG/T |
| Electron Gun | FIELD EMISSION GUN |
| Accelerating Voltage (kV) | 300 |
| Cs (mm) | 2 |
| Detector | KODAK SO-163 FILM |
3D Reconstruction
| Method | SINGLE PARTICLE |
| Resolution (Å) | 10.3 |
| Software | EMfit,Auto3DEM |
| Symmetry | I |
