Subtomogram averaging from cryo-electron tomograms.
Details
Serval ID
serval:BIB_2DC445459696
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
Subtomogram averaging from cryo-electron tomograms.
Journal
Methods in cell biology
ISSN
0091-679X (Print)
ISSN-L
0091-679X
Publication state
Published
Issued date
2019
Peer-reviewed
Oui
Volume
152
Pages
217-259
Language
english
Notes
Publication types: Journal Article ; Research Support, Non-U.S. Gov't ; Review
Publication Status: ppublish
Publication Status: ppublish
Abstract
Cryo-electron tomography (cryo-ET) allows three-dimensional (3D) visualization of frozen-hydrated biological samples, such as protein complexes and cell organelles, in near-native environments at nanometer scale. Protein complexes that are present in multiple copies in a set of tomograms can be extracted, mutually aligned, and averaged to yield a signal-enhanced 3D structure up to sub-nanometer or even near-atomic resolution. This technique, called subtomogram averaging (StA), is powered by improvements in EM hardware and image processing software. Importantly, StA provides unique biological insights into the structure and function of cellular machinery in close-to-native contexts. In this chapter, we describe the principles and key steps of StA. We briefly cover sample preparation and data collection with an emphasis on image processing procedures related to tomographic reconstruction, subtomogram alignment, averaging, and classification. We conclude by summarizing current limitations and future directions of this technique with a focus on high-resolution StA.
Keywords
Cryoelectron Microscopy/methods, Electron Microscope Tomography/methods, Image Processing, Computer-Assisted/methods, Imaging, Three-Dimensional/methods, Software, Algorithms, Classification, Contrast transfer function, Cryo-electron tomography, Image processing, Sub-nanometer resolution, Subtomogram averaging, Tilt series alignment, Tomogram reconstruction
Pubmed
Web of science
Create date
22/07/2024 11:25
Last modification date
23/07/2024 6:57