ArticleNature communications2024
Overcoming resolution attenuation during tilted cryo-EM data collection.
Article in Nature communications, 2024. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 17 papers.
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17 citing papers in PubMed.
- Structural Mechanism of Filamentation Induced Dampening of GTP Inhibition of Glutamate Dehydrogenase.bioRxiv : the preprint server for biology · 2026Article
- Structural basis of transcription of the hachimoji eight-letter alphabet by E. coli RNA polymerase.Nature communications · 2026Article
- Integrating multitemperature sample preparation with particle rebalancing addresses preferred orientation.PNAS nexus · 2026Article
- Two datasets are better than one: method of double moments for 3D reconstruction in cryo-EM.Inverse problems · 2026Article
- UsingbioRxiv : the preprint server for biology · 2026Article
- Atomic resolution cryo-EM at 200 keV.IUCrJ · 2026Article
- Cryo-EM Structures Reveal the Molecular Basis of Asymmetric Allosteric Activation by MMOB in the Hydroxylase of Soluble Methane Monooxygenase.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2026Article
- A Surfactant Cocktail Overcomes Air-Water Interface Artifacts in Single-Particle CryoEM.bioRxiv : the preprint server for biology · 2026Article
- Scaffolds with optimized quaternary symmetry for de novo cryoEM structure determination of small RNAs.Nature methods · 2026Article
- Oligomeric HIV-1 integrase structures reveal functional plasticity for intasome assembly and RNA binding.Nature communications · 2025Article
- How many (distinguishable) classes can we identify in single-particle analysis?Acta crystallographica. Section D, Structural biology · 2025Article
- Alternatives for Uranyl Acetate Negative Staining With Respect to the Resolution Obtained After Single Particle Analysis of Erythrocruorin.Microscopy research and technique · 2025Article
- Laser flash melting cryo-EM samples to overcome preferred orientation.Nature methods · 2025Article
- CryoPROS: Correcting misalignment caused by preferred orientation using AI-generated auxiliary particles.Nature communications · 2025Article
- Overcoming the preferred-orientation problem in cryo-EM with self-supervised deep learning.Nature methods · 2025Article
- Factors affecting macromolecule orientations in thin films formed in cryo-EM.Acta crystallographica. Section D, Structural biology · 2024Article
- Overcoming the preferred orientation problem in cryoEM with self-supervised deep-learning.bioRxiv : the preprint server for biology · 2024Article
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22 authors.
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Abstract
Structural biology efforts using cryogenic electron microscopy are frequently stifled by specimens adopting "preferred orientations" on grids, leading to anisotropic map resolution and impeding structure determination. Tilting the specimen stage during data collection is a generalizable solution but has historically led to substantial resolution attenuation. Here, we develop updated data collection and image processing workflows and demonstrate, using multiple specimens, that resolution attenuation is negligible or significantly reduced across tilt angles. Reconstructions with and without the stage tilted as high as 60° are virtually indistinguishable. These strategies allowed the reconstruction to 3 Å resolution of a bacterial RNA polymerase with preferred orientation, containing an unnatural nucleotide for studying novel base pair recognition. Furthermore, we present a quantitative framework that allows cryo-EM practitioners to define an optimal tilt angle during data acquisition. These results reinforce the utility of employing stage tilt for data collection and provide quantitative metrics to obtain isotropic maps.
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