FBS Colloquia No.264JEOL YOKOGUSHI Research Alliance Laboratories

Fast! Easy! Development of high-throughput CryoEM imaging systems and new type grid using CRYO ARM 300

Structural analysis using CryoEM does not require crystallization of the sample, and data collection can be performed with only a few μl of sample. Recently, the resolution has become as high as the X-ray crystallography.
At the Graduate School of FBS, we have built a high-throughput automated data acquisition system based on the CRYO ARM 300 provided by JEOL, which enables us to acquire up to 1000 images per hour. At the same time, we have built the system that can analyze the data in real time and make decisions using AI. It is also one of the world's leading CryoEM facilities, with a full support environment, including manuals and members of the Namba Lab providing support during operations, and advice about grids.
In collaboration with the research group of Inoue at the Graduate School of Pharmaceutical Sciences, Osaka University, we have successfully applied the oxidation reaction using photoactivated ClO2 gas, which can convert methane into methanol, to the graphene grid and named it EG-grid. The EG-grid is an ideal grid that avoids the denaturation problem caused by the air-water interface. Using this grid, we have achieved resolutions of 1.98 Å, 1.83 Å, and 1.29 Å for three proteins, GroEL, β-galactosidase, and apoferritin, which are benchmarks in CryoEM structural analysis, respectively. This EG-grid will be a powerful tool to accelerate high-resolution structural analysis in CryoEM. In this seminar, we will introduce each of them.