Visualizing Biomolecules through High-Resolution Cryo-EM

The cryogenic electron microscopy (cryo-EM) field exploded in popularity during the 2010s' "resolution revolution."

The 2017 Nobel Prize in Chemistry awarded Jacques Dubochet, Joachim Frank, and Richard Henderson "for developing cryo-electron microscopy for the high-resolution structure determination of biomolecules in solution."

Cryo-EM techniques, such as single particle analysis (SPA), now enable structural identification of biomolecules at resolutions previously only possible with X-ray crystallography.

Cryo-EM opens up many possibilities because it does not require protein crystallization and can determine the structural composition of massive protein complexes observed in frozen solutions or cellular environments.

However, there are still many challenges in adopting cryo-EM; for example, the screening procedure to distinguish good samples from poor ones demands efficiency because microscopy time is becoming increasingly rare due to the constant influx of new users.

Due to their intrinsic flexibility and dynamic nature, structural investigations of protein complexes that orchestrate dynamic biological processes are frequently limited. This necessitates high-throughput data-gathering methodologies followed by extensive 2D categorization algorithms.

Structure-based drug design with cryo-EM necessitates high-resolution/high-throughput imaging workflows to generate density maps that can reliably detect a drug-binding pose. Proteins that are too small require strategies that improve contrast for particle selection.

As a result, cryo-EM technologies and workflows that enable rapid screening, processing, and data collection are becoming increasingly crucial, particularly in efforts to solve important but difficult structures.

To address the problems of the growing cryo-EM industry, JEOL released the CRYO ARM 200 and 300 in 2017. These were JEOL's first 200 kV and 300 kV dedicated cryo-transmission electron microscopes.

 

 

Image Credit: JEOL USA, Inc.

About the Presenters

Emmanuel Smith, Senior Application Specialist for Cryo-EM at JEOL USA

Emmanuel works as a cryo-EM Applications Specialist for JEOL USA, providing user assistance for the JEOL CRYO ARM microscopes throughout North America. His background is in structural biology. He earned his PhD in X-ray crystallography and structure-based drug design from Yu Chen's lab at the University of South Florida.

He completed his postdoctoral research in cryo-EM at Daniela Rhodes' group at Nanyang Technological University, where he focused on telomere-binding protein complexes.

He eventually worked as a staff member at two cryo-EM facilities before returning to research in Tina Izard's lab at the Scripps Research Institute Florida campus, where he studied F-actin protein complexes with the first JEOL CRYO ARM 300 in the US.

He is familiar with all areas of the SPA procedure, including sample preparation, data collecting, and analysis.

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