Super-Resolution Processing Enables In Situ Subtomogram Averaging Beyond the Nyquist Limit
- 1EMBL
Abstract
In situ subtomogram averaging (STA) for high-resolution structure determination requires fine pixel sampling, forcing a trade-off between achievable resolution and the field of view (FOV) recorded in each tomogram. Electron-event representation (EER) data can be rendered at half the physical pixel size, but whether super-resolution processing can recover useful information beyond the physical Nyquist limit from noisy in situ data remains unclear. Here, we show that super-resolution processing of EER data recovers biologically interpretable information beyond a physical Nyquist limit of 4.78 A in two in situ datasets. STA of unidentified virus particles in a Drosophila melanogaster larva yielded a 3.42 A inner capsid reconstruction, enabling de novo model building and identification of the particles as Drosophila A virus. In situ STA of ribosomes in Saccharomyces cerevisiae reached 3.18 A, with local resolution extending to 2.78 A. EER acquisition therefore provides greater flexibility in choosing the acquisition magnification: existing datasets can be re-rendered at the super-resolution pixel size when physical sampling becomes limiting, while lower-magnification acquisition can be considered when a larger FOV improves target capture or cellular context.
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BibTeX
@misc{pyle2026superresolution,
title = {{Super-Resolution Processing Enables In Situ Subtomogram Averaging Beyond the Nyquist Limit}},
author = {E. Pyle and S. Bekbayev and V. Dmello and S. Mattei},
howpublished = {bioRxiv},
year = {2026},
month = oct,
doi = {10.64898/2026.10.17.759810},
url = {https://www.biorxiv.org/content/10.64898/2026.10.17.759810v1},
note = {CC BY},
}
RIS
TY - UNPB
TI - Super-Resolution Processing Enables In Situ Subtomogram Averaging Beyond the Nyquist Limit
AU - E. Pyle
AU - S. Bekbayev
AU - V. Dmello
AU - S. Mattei
PY - 2026
DA - 2026/10/17
PB - bioRxiv
DO - 10.64898/2026.10.17.759810
UR - https://www.biorxiv.org/content/10.64898/2026.10.17.759810v1
AB - In situ subtomogram averaging (STA) for high-resolution structure determination requires fine pixel sampling, forcing a trade-off between achievable resolution and the field of view (FOV) recorded in each tomogram. Electron-event representation (EER) data can be rendered at half the physical pixel size, but whether super-resolution processing can recover useful information beyond the physical Nyquist limit from noisy in situ data remains unclear. Here, we show that super-resolution processing of EER data recovers biologically interpretable information beyond a physical Nyquist limit of 4.78 A in two in situ datasets. STA of unidentified virus particles in a Drosophila melanogaster larva yielded a 3.42 A inner capsid reconstruction, enabling de novo model building and identification of the particles as Drosophila A virus. In situ STA of ribosomes in Saccharomyces cerevisiae reached 3.18 A, with local resolution extending to 2.78 A. EER acquisition therefore provides greater flexibility in choosing the acquisition magnification: existing datasets can be re-rendered at the super-resolution pixel size when physical sampling becomes limiting, while lower-magnification acquisition can be considered when a larger FOV improves target capture or cellular context.
ER -