Energy filtering enables macromolecular MicroED data at sub-atomic resolution

Abstract High-resolution information is important for accurate structure modeling but is challenging to attain in macromolecular crystallography due to the rapid fading of diffracted intensities at increasing resolution. While direct electron detection essentially eliminates the read-out noise durin...

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Main Authors: Max T. B. Clabbers, Johan Hattne, Michael W. Martynowycz, Tamir Gonen
Format: Article
Language:English
Published: Nature Portfolio 2025-03-01
Series:Nature Communications
Online Access:https://doi.org/10.1038/s41467-025-57425-1
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author Max T. B. Clabbers
Johan Hattne
Michael W. Martynowycz
Tamir Gonen
author_facet Max T. B. Clabbers
Johan Hattne
Michael W. Martynowycz
Tamir Gonen
author_sort Max T. B. Clabbers
collection DOAJ
description Abstract High-resolution information is important for accurate structure modeling but is challenging to attain in macromolecular crystallography due to the rapid fading of diffracted intensities at increasing resolution. While direct electron detection essentially eliminates the read-out noise during MicroED data collection, other sources of noise remain and limit the measurement of faint high-resolution reflections. Inelastic scattering significantly contributes to noise, raising background levels and broadening diffraction peaks. We demonstrate a substantial improvement in signal-to-noise ratio by using energy filtering to remove inelastically scattered electrons. This strategy results in sub-atomic resolution MicroED data from proteinase K crystals, enabling the visualization of detailed structural features. Interestingly, reducing the noise further reveals diffuse scattering that may hold additional structural information. Our findings suggest that combining energy filtering and direct detection provides more accurate measurements at higher resolution, facilitating precise model refinement and improved insights into protein structure and function.
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institution DOAJ
issn 2041-1723
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publishDate 2025-03-01
publisher Nature Portfolio
record_format Article
series Nature Communications
spelling doaj-art-b8659a559da942bf949ca3f58dadf0682025-08-20T02:52:19ZengNature PortfolioNature Communications2041-17232025-03-011611610.1038/s41467-025-57425-1Energy filtering enables macromolecular MicroED data at sub-atomic resolutionMax T. B. Clabbers0Johan Hattne1Michael W. Martynowycz2Tamir Gonen3Howard Hughes Medical Institute, University of CaliforniaHoward Hughes Medical Institute, University of CaliforniaHoward Hughes Medical Institute, University of CaliforniaHoward Hughes Medical Institute, University of CaliforniaAbstract High-resolution information is important for accurate structure modeling but is challenging to attain in macromolecular crystallography due to the rapid fading of diffracted intensities at increasing resolution. While direct electron detection essentially eliminates the read-out noise during MicroED data collection, other sources of noise remain and limit the measurement of faint high-resolution reflections. Inelastic scattering significantly contributes to noise, raising background levels and broadening diffraction peaks. We demonstrate a substantial improvement in signal-to-noise ratio by using energy filtering to remove inelastically scattered electrons. This strategy results in sub-atomic resolution MicroED data from proteinase K crystals, enabling the visualization of detailed structural features. Interestingly, reducing the noise further reveals diffuse scattering that may hold additional structural information. Our findings suggest that combining energy filtering and direct detection provides more accurate measurements at higher resolution, facilitating precise model refinement and improved insights into protein structure and function.https://doi.org/10.1038/s41467-025-57425-1
spellingShingle Max T. B. Clabbers
Johan Hattne
Michael W. Martynowycz
Tamir Gonen
Energy filtering enables macromolecular MicroED data at sub-atomic resolution
Nature Communications
title Energy filtering enables macromolecular MicroED data at sub-atomic resolution
title_full Energy filtering enables macromolecular MicroED data at sub-atomic resolution
title_fullStr Energy filtering enables macromolecular MicroED data at sub-atomic resolution
title_full_unstemmed Energy filtering enables macromolecular MicroED data at sub-atomic resolution
title_short Energy filtering enables macromolecular MicroED data at sub-atomic resolution
title_sort energy filtering enables macromolecular microed data at sub atomic resolution
url https://doi.org/10.1038/s41467-025-57425-1
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AT johanhattne energyfilteringenablesmacromolecularmicroeddataatsubatomicresolution
AT michaelwmartynowycz energyfilteringenablesmacromolecularmicroeddataatsubatomicresolution
AT tamirgonen energyfilteringenablesmacromolecularmicroeddataatsubatomicresolution