Cryo-Electron Microscopy: The field of 1,000⁺ methods

Area of Science:

• Structural Biology
• Biophysics
• Computational Biology

Background:

• Cryo-Electron Microscopy (CryoEM) is a powerful technique for determining the 3D structures of biological macromolecules.
• Advances in sample preparation, electron optics, detection, image acquisition, processing, and interpretation have fueled CryoEM's success.
• The field has seen continuous methodological development since the 1960s, with a recent surge in publications.

Purpose of the Study:

• To analyze trends in CryoEM methodological publications, focusing on computational image processing.
• To identify key areas of development and potential confusion for practitioners.
• To highlight the importance of best practices in methods reporting and usage.

Main Methods:

• Systematic review and trend analysis of over 1,000 methodological papers in CryoEM.
• Categorization of papers by CryoEM technique (e.g., Single Particle Analysis, Electron Tomography) and methodological aspect (e.g., sample prep, image processing, reconstruction).
• Focus on computational image processing methods, while also considering sample preparation and image acquisition.

Main Results:

• Single Particle Analysis (SPA) has experienced significant acceleration in the last decade, with sample preparation methods advancing rapidly in the last five years.
• Electron Tomography is poised for future growth.
• 3D classification, 3D reconstruction, and 3D alignment remain critical, actively evolving areas within SPA.
• Atomic modeling has emerged as a significant research focus since the resolution revolution.

Conclusions:

• The rapid development of CryoEM methods, particularly in computational image processing, necessitates clear best practices for reporting and usage.
• Understanding these trends is crucial for researchers to effectively apply and advance CryoEM techniques.
• Continued innovation in SPA, Electron Tomography, and atomic modeling promises further breakthroughs in structural biology.