Loading...
Thumbnail Image
Publication

Defocus Corrected Large Area Cryo-EM (DeCo-LACE) for label-free detection of molecules across entire cell sections

Elferich, Johannes
Schiroli, Giulia
Scadden, David T
Grigorieff, Nikolaus
Citations
Altmetric:
Student Authors
Faculty Advisor
Academic Program
UMass Chan Affiliations
Document Type
Journal Article
Publication Date
2022-11-16
Subject Area
Embargo Expiration Date
Abstract

A major goal of biological imaging is localization of biomolecules inside a cell. Fluorescence microscopy can localize biomolecules inside whole cells and tissues, but its ability to count biomolecules and accuracy of the spatial coordinates is limited by the wavelength of visible light. Cryo-electron microscopy (cryo-EM) provides highly accurate position and orientation information of biomolecules but is often confined to small fields of view inside a cell, limiting biological context. In this study, we use a new data-acquisition scheme called Defocus-Corrected Large-Area cryo-EM (DeCo-LACE) to collect high-resolution images of entire sections (100- to 250-nm-thick lamellae) of neutrophil-like mouse cells, representing 1-2% of the total cellular volume. We use 2D template matching (2DTM) to determine localization and orientation of the large ribosomal subunit in these sections. These data provide maps of ribosomes across entire sections of mammalian cells. This high-throughput cryo-EM data collection approach together with 2DTM will advance visual proteomics and provide biological insight that cannot be obtained by other methods.

Source

Elferich J, Schiroli G, Scadden DT, Grigorieff N. Defocus Corrected Large Area Cryo-EM (DeCo-LACE) for label-free detection of molecules across entire cell sections. Elife. 2022 Nov 16;11:e80980. doi: 10.7554/eLife.80980. PMID: 36382886; PMCID: PMC9711527.

Year of Medical School at Time of Visit
Sponsors
Dates of Travel
DOI
10.7554/eLife.80980
PubMed ID
36382886
Other Identifiers
Notes
Funding and Acknowledgements
Corresponding Author
Related Resources
Related Resources
Repository Citation
Rights
Copyright Elferich et al. This article is distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use and redistribution provided that the original author and source are credited.Attribution 4.0 International