Expanding the Frontiers of Cryo-EM Towards More Challenging Targets

The 2023 GRC on Three Dimensional Electron Microscopy will continue its tradition of spotlighting cutting-edge results and recent methods developments in a highly interactive and open-minded environment.

Through numerous technical advances in the past decade, cryo-EM single particle analysis matured into a powerful and reliable method for protein structure determination, and it has become an indispensable part of the structural methods toolbox. Software developments in data processing and experiment automation improved the quality of results and the accessibility of the technique. Instrumentation advances increased throughput and improved data quality. Nevertheless, there are still numerous challenges that hold opportunities for future advances. Sample preparation remains the most unpredictable step in the cryo-EM workflow and needs further quantification and development. Deep learning-based automation of sample screening, data acquisition, and data processing hold promise for improved ease of use and efficiency. Small (<50 kDa), multi-component, heterogeneous, flexible, or intermittent complexes provide deeper biological insights but remain very challenging.

Cryo-electron tomography saw substantial growth in the last few years, both in resolution and accessibility. Together with high-resolution template matching, it is in pursuit of the ultimate goal in structural biology - atomic resolution observation of protein complexes in their native unperturbed cellular environment. However, there are many obstacles on the road to “in situ visual proteomics”. Like in single particle analysis, sample preparation is a key factor that requires more development. Accurate targeting and identification of areas of interest are not sufficiently reliable. Cryo-focused ion beam (cryo-FIB) sample preparation yield, throughput, and automation have room for improvement. Automated tomography data collection and (on-the-fly) processing are also areas with great potential for future advances.

For all experimental approaches, the optical performance, reliability, ease of use, and maintenance costs of the equipment are of fundamental importance and will be discussed.

Due to the growing popularity of cryo-EM, the meeting has been heavily oversubscribed in recent years and therefore early applications are encouraged. Please provide an abstract emphasizing the unique and novel aspects of your work, and for junior members of the community, please include the name of your advisor/PI.