Single-particle cryo-EM structures from iDPC–STEM at near-atomic resolution

Autor: 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
Jazyk: angličtina
Rok vydání: 2022
Předmět:
Zdroj: Nature methods 1, 38 (2022). doi:10.1038/s41592-022-01586-0
Popis: 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.
Databáze: OpenAIRE