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TitleSingle-particle cryo-EM structures from iDPC-STEM at near-atomic resolution.
Journal, issue, pagesNat Methods, Vol. 19, Issue 9, Page 1126-1136, Year 2022
Publish dateSep 5, 2022
AuthorsIvan 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 AbstractIn 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 linksNat Methods / PubMed:36064775 / PubMed Central
MethodsEM (helical sym.) / EM (single particle)
Resolution3.5 - 6.51 Å
Structure data

13778

7q22

EMDB-13778, PDB-7q22:
cryo iDPC-STEM structure recorded with CSA 2.0
Method: EM (helical sym.) / Resolution: 6.3 Å

13779

7q23

EMDB-13779, PDB-7q23:
cryo iDPC-STEM structure recorded with CSA 3.0
Method: EM (helical sym.) / Resolution: 4.3 Å

13780

7q2q

EMDB-13780, PDB-7q2q:
cryo iDPC-STEM structure recorded with CSA 3.5
Method: EM (helical sym.) / Resolution: 4.3 Å

13781

7q2r

EMDB-13781, PDB-7q2r:
cryo iDPC-STEM structure recorded with CSA 4.0
Method: EM (helical sym.) / Resolution: 3.5 Å

13782

7q2s

EMDB-13782, PDB-7q2s:
cryo iDPC-STEM structure recorded with CSA 4.5
Method: EM (helical sym.) / Resolution: 3.7 Å

EMDB-14407: Single particle structure of keyhole limpet hemocyanin obtained via iDPC scanning transmission electron microscopy
Method: EM (single particle) / Resolution: 6.51 Å

Source
  • tobacco mosaic virus (strain vulgare)
  • tobacco mosaic virus (vulgare)
  • TMV, Tobacco mosaic virus (strain U1)
  • Megathura crenulata (invertebrata)
KeywordsVIRUS / tobacco mosaic virus / RNA virus

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