+Search query
-Structure paper
Title | Single-particle cryo-EM structures from iDPC-STEM at near-atomic resolution. |
---|---|
Journal, issue, pages | Nat Methods, Vol. 19, Issue 9, Page 1126-1136, Year 2022 |
Publish date | Sep 5, 2022 |
Authors | Ivan Lazić / Maarten Wirix / Max Leo Leidl / Felix de Haas / Daniel Mann / Maximilian Beckers / Evgeniya V Pechnikova / Knut Müller-Caspary / Ricardo Egoavil / Eric G T Bosch / Carsten Sachse / |
PubMed Abstract | In electron cryomicroscopy (cryo-EM), molecular images of vitrified biological samples are obtained by conventional transmission microscopy (CTEM) using large underfocuses and subsequently ...In electron cryomicroscopy (cryo-EM), molecular images of vitrified biological samples are obtained by conventional transmission microscopy (CTEM) using large underfocuses and subsequently computationally combined into a high-resolution three-dimensional structure. Here, we apply scanning transmission electron microscopy (STEM) using the integrated differential phase contrast mode also known as iDPC-STEM to two cryo-EM test specimens, keyhole limpet hemocyanin (KLH) and tobacco mosaic virus (TMV). The micrographs show complete contrast transfer to high resolution and enable the cryo-EM structure determination for KLH at 6.5 Å resolution, as well as for TMV at 3.5 Å resolution using single-particle reconstruction methods, which share identical features with maps obtained by CTEM of a previously acquired same-sized TMV data set. These data show that STEM imaging in general, and in particular the iDPC-STEM approach, can be applied to vitrified single-particle specimens to determine near-atomic resolution cryo-EM structures of biological macromolecules. |
External links | Nat Methods / PubMed:36064775 / PubMed Central |
Methods | EM (helical sym.) / EM (single particle) |
Resolution | 3.5 - 6.51 Å |
Structure data | EMDB-13778, PDB-7q22: EMDB-13779, PDB-7q23: EMDB-13780, PDB-7q2q: EMDB-13781, PDB-7q2r: EMDB-13782, PDB-7q2s: EMDB-14407: Single particle structure of keyhole limpet hemocyanin obtained via iDPC scanning transmission electron microscopy |
Source |
|
Keywords | VIRUS / tobacco mosaic virus / RNA virus |