Abstract
Cryo-electron microscopy (Cryo-EM) has become a routine technology for resolving the structure of biological macromolecules due to the resolution revolution in recent years. The specimens are typically prepared in a very thin layer of vitrified ice suspending in the holes of the perforated amorphous carbon film. However, the samples prepared by directly applying to the conventional support membranes may suffer from partial or complete denaturation caused by sticking to the air–water interface (AWI). With the application in materials, graphene has also been used recently to improve frozen sample preparation instead of a suspended conventional amorphous thin carbon. It has been proven that graphene or graphene oxide and various chemical modifications on its surface can effectively prevent particles from adsorbing to the AWI, which improves the dispersion, adsorbed number, and orientation preference of frozen particles in the ice layer. Their excellent properties and thinner thickness can significantly reduce the background noise, allowing high-resolution three-dimensional reconstructions using a minimum data set.
Highlights
We mainly focus on the characteristics of graphene and graphene oxide (GO), the recently proposed preparation procedure of graphene or GO suspended EM grid, the derivative surface modifications, and the related benefits to cryo-EM sample preparation
The addition of stabilizers and surfactants can diminish the signal-to-noise due to the air–water interface (AWI) can happen within milliseconds or even less [71,72,74,75,76], which can be ratio (SNR) dramatically, especially disastrous for the small molecules, and change the summarized in three stages [77]: first, protein adsorbs to the AWI, followed by shearing surface tension of the solution and the sample preparation conditions
The graphene or GO support has many excellent properties, whether to use them depends on the target protein or complex
Summary
Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affiliations. It has been recently reported that the cryo-EM single-particle technique can achieve a resolution as high as 1.22 Å [22], the quality of structure appears better than that obtained at the same level of resolution by X-ray crystal diffraction [23]; this may not be universally achievable. It mostly depends on the property of the macromolecule itself, such as the intrinsic stability and its behavior in the specimen preparation. We mainly focus on the characteristics of graphene and GO, the recently proposed preparation procedure of graphene or GO suspended EM grid, the derivative surface modifications, and the related benefits to cryo-EM sample preparation
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