Cryo Grid Box Screw Type Lid Transfer: A Solution for Overcoming Preferential Orientation Problem in Cryo-EM Specimen Preparation

Cryogenic electron microscopy (cryo-EM) has revolutionized the field of structural biology by enabling researchers to determine the three-dimensional structures of biological macromolecules at atomic resolution. However, achieving high-resolution structures is often hindered by severe preferential orientation problems on conventional EM grids. To address this challenge, scientists have developed modified graphene materials with different charge properties that enable particle adsorption on the surface with favorable orientation distribution.

In a recent breakthrough, researchers successfully determined the cryo-EM structure of Ll.LtrB RNP at 3.2-Å resolution using a novel method for producing robust graphene grids functionalized with different electrostatic charges. This achievement overcomes the limitations of conventional EM grids and provides new opportunities for high-resolution structure determination.

The development of clean graphene grids is crucial for cryo-EM specimen preparation, as contamination built-up on the graphene surface during the transfer process or storage can hinder the effectiveness of bioactive functionalization processes. Furthermore, graphene membrane, an atomically thin film comprised of sp2-bonded carbon atoms, has been shown to induce preferential orientation problems in particles adsorbed to its surface.

To alleviate these challenges, researchers have introduced supporting films, including graphene membranes, which are capable of mitigating the preferential orientation problem. Additionally, tilting the cryo-specimen during data collection has been applied as a solution, but this method can result in large beam-induced motion and inaccurate defocus estimation during data processing.

In summary, the development of cryo grid box screw type lid transfer technology offers a promising solution for overcoming the preferential orientation problem in cryo-EM specimen preparation. By providing various electrostatic interaction interfaces for target macromolecules to bind in and enrich orientations, this technology has the potential to revolutionize our understanding of biological processes at the molecular level.

Cryo grid box screw type lid transfer