A Primer to Single-Particle Cryo-Electron Microscopy

By Yifan Cheng, N. Grigorieff, Pawel A. Penczek, Thomas Walz. 2015

527 citations recorded by the source corpus.

Summary Cryo-electron microscopy (cryo-EM) of single-particle specimens is used to determine the structure of proteins and macromolecular complexes without the need for crystals. Recent advances in detector technology and software algorithms now allow images of unprecedented quality to be recorded and structures to be determined at near-atomic resolution. However, compared with X-ray crystallography, cryo-EM is a young technique with distinct challenges. This primer explains the different steps and considerations involved in structure determination by single-particle cryo-EM to provide an overview for scientists wishing to understand more about this technique and the interpretation of data obtained with it, as well as a starting guide for new practitioners.

Subfields: Materials Chemistry, Molecular Biology, Structural Biology

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Citations in the corpus

Measuring the optimal exposure for single particle cryo-EM using a 2.6 Å reconstruction of rotavirus VP6 (2015, background): “…number of structures determined at near-atomic resolution (Liao et al., 2013; Allegretti et al., 2014; Bartesaghi et al., 2014; Wong et al., 2014), a level of detail that had previously been restricted to X-ray crystallography and NMR (for a recent review of the technique, see Cheng et al., 2015).”
Vitrification after multiple rounds of sample application and blotting improves particle density on cryo-electron microscopy grids (2016, unknown): “Single particle cryo-electron microscopy (cryoEM) is becoming an increasingly important tool in structural biology (Nogales and Scheres 2015; Cheng et al. 2015).”
DeepPicker: a Deep Learning Approach for Fully Automated Particle Picking in Cryo-EM (2016, background): “Recent advance of the single-particle electron cryo-microscopy (cryo-EM) technique has been revolutionizing the structural biology ﬁeld [1, 2, 3], and enabled protein complex structure determination at near-atomic resolution [4, 5, 6, 7].”
Accelerated cryo-EM structure determination with parallelisation using GPUs in RELION-2 (2016, methodology): “This is achieved by isolating two-dimensional particle-projections in the micrographs, which can then be recombined into a three-Kimanius dimensional structure ( Cheng et al., 2015 ).”