RSB An Introduction to CryoEM

Within our cells, biological molecules such as proteins, DNA and RNA are organised into dynamic complexes. Structural biology aims to understand how these complexes assemble and function as molecular machines. Cryo electron microscopy (cryoEM) allows us to study these molecular machines in a frozen hydrated state, preserving them close to their natural environment.

This course provides an introduction to cryoEM and its applications in structural biology. You will learn what types of samples are suitable for cryoEM, what results can be expected, and the steps involved in generating high quality structural data. As part of the training, you will visit and get experience using equipment in the Midlands Regional CryoEM Facility.

The course will focus on single particle analysis (SPA), a computational method that aligns and averages thousands to millions of individual transmission electron microscope (TEM) images. This process produces a 3D cryoEM map with sufficient detail to reveal how proteins function and how they interact with other molecules, including potential drug compounds.

Aims

The course aims to provide participants with an introductory understanding of cryoEM by the following objectives:

• Gain an understanding of sample preparation and imaging using negative-stain TEM.
• Obtain experience with vitrification equipment used for cryoEM sample preparation.
• Get hands-on experience operating an electron microscope for cryoTEM imaging and data collection.
• Learn the fundamental principles of single-particle analysis, including how 2D projection images are processed and combined to generate a 3D structure.