Making Graphene Oxide Grid¶
This protocol is based on MRC LMB one. We modify the protocol based on Xudong Wu’s method at Harvard Medical School.
Materials and Equipment Used:¶
GO Solution (Sigma:763705, 2 mg/ml)
Pipette (1 µl, 3 µl and 20 µl)
Whatman filter paper No.1
Dilute GO solution 10x to 0.2 mg/mL with ddH2O
Sonicate for 15s to break up aggregates
Spin for 30 seconds at 300 rcf to remove aggregates
Prepare a flat and clean working area (e.g. with parafilm)
Further dilute GO solution 10x to 0.02 mg/mL with ddH2O
Glow discharge grids with carbon side up (easiGlow (PELCO, Discharge System); Quantifoil Cu 300 R1.2/1.3 120 seconds at 0.2 mBar and 15 mA)
Take up grids with anti-capillary tweezers
Place 1µl of the diluted GO solution (4) on the carbon side of the grid
Wait for the grid to dry out
This method is good for high coverage of GO, but maybe get multiple layers of GO on the grid.
Discharging conditions are varied between grids and discharge system.