Fabrication of Monolayer Graphene-Coated Grids for Cryoelectron Microscopy

Currently optimally preserving biological samples is a major challenge in obtaining high resolution CryoEM structures. During the sample preparation process, specimens are exposed to hydrophobic air-water interface, which has denaturing effects on the samples and can cause stable samples to adopt a preferential orientation in the ice, which prevents high resolution structure determination. Coating CryoEM grids with a thin layer of carbon of like soft graphene oxide, has been commonly used to address issues related to interactions with air-water interface.

However, this layers often increase background noise in our images, which compromise the resolutions of our structure. Coating the grid with a monolayer of graphene accomplishes similar protection without introducing significant background noise. Using graphene-coated CryoEM grids can help overcome issues related to interactions with air-water interface, however, commercial graphene grids are expensive and many labs consider graphene grids to be too complicated to prepare in-house.

Researchers in the CryoEM field can now use this step-by-step protocol to reproduce fully produced dozens of high quality graphenes in one day.