Making Graphene Oxide Grid

Author

Kangkang Song

Contact

<kangkang.song@umassmed.edu>

Date-created

2018-3-10

Last-updated

2018-8-21

Abstract

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:

1. GO Solution (Sigma:763705, 2 mg/ml)

2. ddH₂O

3. Tweezers

4. Pipette (1 µl, 3 µl and 20 µl)

5. Tabletop centrifuge

6. Parafilm

7. Whatman filter paper No.1

Procedure:

1. Dilute GO solution 10x to 0.2 mg/mL with ddH₂O

2. Sonicate for 15s to break up aggregates

3. Spin for 30 seconds at 300 rcf to remove aggregates

4. Prepare a flat and clean working area (e.g. with parafilm)

5. Further dilute GO solution 10x to 0.02 mg/mL with ddH₂O

6. 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)

7. Take up grids with anti-capillary tweezers

8. Place 1µl of the diluted GO solution (4) on the carbon side of the grid

9. Wait for the grid to dry out

Note

1. This method is good for high coverage of GO, but maybe get multiple layers of GO on the grid.

2. Discharging conditions are varied between grids and discharge system.