Prospects and Limitations of High-Resolution Single-Particle Cryo-Electron Microscopy

Single particle cryo-electron microscopy (cryo-EM) has matured into a robust method for the determination of biological macromolecule structures in the past decade, complementing X-ray crystallography and nuclear magnetic resonance. Constant methodological improvements in both cryo-EM hardware and image processing software continue to contribute to an exponential growth in the number of structures solved annually. In this review, we provide a historical view of the many steps that were required to make cryo-EM a successful method for the determination of high-resolution protein complex structures. We further discuss aspects of cryo-EM methodology that are the greatest pitfalls challenging successful structure determination to date. Lastly, we highlight and propose potential future developments that would improve the method even further in the near future.

Automated summary

Single particle cryo-electron microscopy (cryo-EM) has become a robust method for determining biological macromolecule structures. Continuous improvements in hardware and software have contributed to an increasing number of solved structures. This review discusses the steps in developing and challenges of cryo-EM methodology, as well as potential future developments.