ArticleNature protocols2016
Resolving macromolecular structures from electron cryo-tomography data using subtomogram averaging in RELION.
Article in Nature protocols, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 160 papers.
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Who cites it
160 citing papers in PubMed, 300 citations in OpenAlex.
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- Deep-learning methods for contrast enhancement and artifact reduction in cryo-electron tomography: a systematic analysis of the state of the art and proposed improvements.Acta crystallographica. Section D, Structural biology · 2026Article
- TANGO: Analysis and curation of particles in cryo-electron tomography.Nature communications · 2026Article
- Molecular mechanisms of flotillin complexes in organizing membrane microdomains.Nature communications · 2026Article
- Resolving structural heterogeneity in situ through cryogenic electron tomography.Current opinion in structural biology · 2026Review
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- Fully genetically encoded low-molecular-weight protein tags with defined shapes for direct molecular identification by cryo-electron tomography.bioRxiv : the preprint server for biology · 2026Article
- Isolation and Visualization Fusobacterium nucleatum Microcompartments.Methods in molecular biology (Clifton, N.J.) · 2026Article
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- Template Learning: Deep learning with domain randomization for particle picking in cryo-electron tomography.Nature communications · 2025Article
- High-throughput cryo-electron tomography enables multiscale visualization of the inner life of microbes.Current opinion in structural biology · 2025Review
- CryoDRGN-AI: neural ab initio reconstruction of challenging cryo-EM and cryo-ET datasets.Nature methods · 2025Article
- The postsynaptic density in excitatory synapses is composed of clustered, heterogeneous nanoblocks.The Journal of cell biology · 2025Article
- The SARS-CoV-2 NSP4 T492I mutation promotes double-membrane vesicle formation to facilitate transmission.Virologica Sinica · 2025Article
- A ternary complex of MIPs in the A-tubule of basal bodies and axonemes depends on RIB22 and the EF-hand domain of RIB72A inMolecular biology of the cell · 2025Article
100 more citing papers are in PubMed but not listed here.
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Authors and funding
2 authors at 1 institution in 1 country.
Funding
Abstract
Electron cryo-tomography (cryo-ET) is a technique that is used to produce 3D pictures (tomograms) of complex objects such as asymmetric viruses, cellular organelles or whole cells from a series of tilted electron cryo-microscopy (cryo-EM) images. Averaging of macromolecular complexes found within tomograms is known as subtomogram averaging, and this technique allows structure determination of macromolecular complexes in situ. Subtomogram averaging is also gaining in popularity for the calculation of initial models for single-particle analysis. We describe herein a protocol for subtomogram averaging from cryo-ET data using the RELION software (http://www2.mrc-lmb.cam.ac.uk/relion). RELION was originally developed for cryo-EM single-particle analysis, and the subtomogram averaging approach presented in this protocol has been implemented in the existing workflow for single-particle analysis so that users may conveniently tap into existing capabilities of the RELION software. We describe how to calculate 3D models for the contrast transfer function (CTF) that describe the transfer of information in the imaging process, and we illustrate the results of classification and subtomogram averaging refinement for cryo-ET data of purified hepatitis B capsid particles and Saccharomyces cerevisiae 80S ribosomes. Using the steps described in this protocol, along with the troubleshooting and optimization guidelines, high-resolution maps can be obtained in which secondary structure elements are resolved subtomogram.
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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.