Challenges in Biological Cryo-electron Microscopy

Book excerpt: Developments in cryo-electron microscopy are creating new opportunities within structural biology and there is currently great interest in developing cryo-EM as a core tool for atomic level structural biology. Many structural techniques can give atomic or near atomic level information, but lack the ability to study proteins within a near-native environment, for example within a cellular compartment. Cryo-EM provides this opportunity, but despite the recent massive improvements in single particle cryo-EM, obtaining sub-2Å structural information is still a major challenge. Cryo-electron microscopy has undergone significant developments in microscope design, camera technology and data processing regimes, but there are significant challenges that remain and opportunities to explore, many of which must be tackled by the community as a whole, rather than by individual groups. For example, sample preparation is central to electron microscopy and is currently a significant bottleneck in many experiments, and there are significant problems with ensuring the integrity of the field in terms of dealing with inherently low signal-to-noise images. This volume brings together leading researchers from the UK and the international cryo-electron microscopy community to discuss current developments and new challenges in the field. In this volume the topics covered include: Sample preparation in single particle cryo-EM ; Pushing the limits in single particle cryo-EM ; Tomographic analysis, CLEM ; Map/model validation and machine learning in EM.