Low-dose cryo-electron ptychography of proteins at sub-nanometer resolution

Berk Küçükoğlu, Inayathulla Mohammed, Ricardo C. Guerrero-Ferreira, Stephanie M. Ribet, Georgios Varnavides, Max Leo Leidl, Kelvin Lau, Sergey Nazarov, Alexander Myasnikov, Massimo Kube, Julika Radecke, Carsten Sachse, Knut Müller-Caspary, Colin Ophus and Henning Stahlberg ()
Additional contact information
Berk Küçükoğlu: Rte. de la Sorge
Inayathulla Mohammed: Rte. de la Sorge
Ricardo C. Guerrero-Ferreira: Rte. de la Sorge
Stephanie M. Ribet: Lawrence Berkeley National Laboratory
Georgios Varnavides: Lawrence Berkeley National Laboratory
Max Leo Leidl: Ernst Ruska-Centre for Microscopy and Spectroscopy with Electrons (ER-C-3): Structural Biology
Kelvin Lau: Rte Cantonale
Sergey Nazarov: EPFL VPA DCI-Lausanne
Alexander Myasnikov: EPFL VPA DCI-Lausanne
Massimo Kube: Rte. de la Sorge
Julika Radecke: Rte. de la Sorge
Carsten Sachse: Ernst Ruska-Centre for Microscopy and Spectroscopy with Electrons (ER-C-3): Structural Biology
Knut Müller-Caspary: Butenandstr. 11
Colin Ophus: Lawrence Berkeley National Laboratory
Henning Stahlberg: Rte. de la Sorge

Nature Communications, 2024, vol. 15, issue 1, 1-10

Abstract: Abstract Cryo-transmission electron microscopy (cryo-EM) of frozen hydrated specimens is an efficient method for the structural analysis of purified biological molecules. However, cryo-EM and cryo-electron tomography are limited by the low signal-to-noise ratio (SNR) of recorded images, making detection of smaller particles challenging. For dose-resilient samples often studied in the physical sciences, electron ptychography – a coherent diffractive imaging technique using 4D scanning transmission electron microscopy (4D-STEM) – has recently demonstrated excellent SNR and resolution down to tens of picometers for thin specimens imaged at room temperature. Here we apply 4D-STEM and ptychographic data analysis to frozen hydrated proteins, reaching sub-nanometer resolution 3D reconstructions. We employ low-dose cryo-EM with an aberration-corrected, convergent electron beam to collect 4D-STEM data for our reconstructions. The high frame rate of the electron detector allows us to record large datasets of electron diffraction patterns with substantial overlaps between the interaction volumes of adjacent scan positions, from which the scattering potentials of the samples are iteratively reconstructed. The reconstructed micrographs show strong SNR enabling the reconstruction of the structure of apoferritin protein at up to 5.8 Å resolution. We also show structural analysis of the Phi92 capsid and sheath, tobacco mosaic virus, and bacteriorhodopsin at slightly lower resolutions.

Date: 2024
References: View references in EconPapers View complete reference list from CitEc
Citations:

Downloads: (external link)
https://www.nature.com/articles/s41467-024-52403-5 Abstract (text/html)

Related works:
This item may be available elsewhere in EconPapers: Search for items with the same title.

Export reference: BibTeX RIS (EndNote, ProCite, RefMan) HTML/Text

Persistent link: https://EconPapers.repec.org/RePEc:nat:natcom:v:15:y:2024:i:1:d:10.1038_s41467-024-52403-5

Ordering information: This journal article can be ordered from
https://www.nature.com/ncomms/

DOI: 10.1038/s41467-024-52403-5

Access Statistics for this article

Nature Communications is currently edited by Nathalie Le Bot, Enda Bergin and Fiona Gillespie

More articles in Nature Communications from Nature
Bibliographic data for series maintained by Sonal Shukla () and Springer Nature Abstracting and Indexing ().