Addressing Preferred Specimen Orientation in Single-Particle Cryo-EM through Tilting
By Y. Tan, Philip Baldwin, Joseph H. Davis, James R. Williamson, Clinton S. Potter, Bridget O. Carragher, D. Lyumkis. 2017
880 citations recorded by the source corpus.
We present a strategy for tackling preferred specimen orientation in single-particle cryogenic electron microscopy by employing tilts during data collection. We also describe a tool to quantify the resulting directional resolution using 3D Fourier shell correlation volumes. We applied these methods to determine the structures at near-atomic resolution of the influenza hemagglutinin trimer, which adopts a highly preferred specimen orientation, and of ribosomal biogenesis intermediates, which adopt moderately preferred orientations.
Subfields: Molecular Biology, Structural Biology, Surfaces, Coatings and Films
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The researchers whose trusted networks rank it highest, strongest advocate first. Every researcher gets a different order.
In Molecular Biology
- Takanori Nakane
- Dari Kimanius
- Erik Lindahl
- Albert J. Kooistra
- Gáspár Pándy-Szekeres
- Jasenko Zivanov
- W. Hagen
- Sjors H. W. Scheres
In Structural Biology
- Dari Kimanius
- Takanori Nakane
- Erik Lindahl
- Björn Forsberg
- W. Hagen
- Jasenko Zivanov
- Sjors H. W. Scheres
- W. Burmeister
In Surfaces, Coatings and Films
- Dari Kimanius
- Erik Lindahl
- Takanori Nakane
- Björn Forsberg
- Jasenko Zivanov
- W. Hagen
- W. Burmeister
- G. Schoehn
Citations in the corpus
- New tools for automated high-resolution cryo-EM structure determination in RELION-3 (2018, methodology): “The latter may occur when the sample support film wrinkles upon flash freezing or when the sample stage is tilted in the microscope, for example to ameliorate a problem of preferred particle orientations ( Tan et al., 2017 ; Naydenova and Russo, 2017 ).”
- 8 Å structure of the outer rings of the Xenopus laevis nuclear pore complex obtained by cryo-EM and AI (2022, methodology): “The validation of the map and model quality of this research was performed using 3D-FSC and Phenix (Afonine et al., 2012; Tan et al., 2017) (Fig.”
- Cryo-EM analysis of a membrane protein embedded in the liposome (2020, methodology): “Directional FSC was calculated by 3DFSC program (50).”
- Structure of the class D GPCR Ste2 dimer coupled to two G proteins (2020, positive): “The directional FSC estimates were calculated using 3DFSC 46 .”
- Transcriptome engineering with RNA-targeting Type VI-D CRISPR effectors (2018, neutral): “Using recently described procedures for characterizing anisotropy in cryo-EM density maps (Tan et al., 2017), we found that the structure maintained approximately even distribution of directional resolution (Figure S1D).”