Whole cryo-FIB lamella acquisition at high magnification with rectangular beam montage cryo-electron tomography
- 1Ian Holmes Imaging Centre, Bio21 Molecular Science and Biotechnology Institute, The University of Melbourne, Parkville, Victoria, Australia
Abstract
The visualisation of large biological samples in cryo-electron tomography requires both few-nanometre resolution and a wide, ideally >10 micrometrre, field of view. For a single image, increasing magnification trades increased resolution for a smaller field of view. Montaging, where high-magnification images are stitched together to increase field of view, addresses the trade-off but is dose-inefficient with round beams that are standard in transmission electron microscopy (TEM), since this illumination shape does not tessellate. Our workflow greatly simplifies montaging schemes for dose-sensitive specimens, allowing large montages of any shape - from individual cells or other features of interest up to whole lamellae - to be collected at high resolution without compromising electron dose efficiency. We demonstrate the workflow by acquiring montage tomograms of whole yeast (Saccharomyces cerevisiae) cells in cryo-FIB lamellae and of human neurons.
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BibTeX
@misc{brown2026whole,
title = {{Whole cryo-FIB lamella acquisition at high magnification with rectangular beam montage cryo-electron tomography}},
author = {H. G. Brown and S. Valimehr and D. Styczynski and M. Mudaliyar and S. S. Kundapura and M. D. Johnson and D. Ghosal and E. Hanssen},
howpublished = {bioRxiv},
year = {2026},
month = sep,
doi = {10.64898/2026.09.27.753993},
url = {https://www.biorxiv.org/content/10.64898/2026.09.27.753993v1},
note = {CC BY},
}
RIS
TY - UNPB
TI - Whole cryo-FIB lamella acquisition at high magnification with rectangular beam montage cryo-electron tomography
AU - H. G. Brown
AU - S. Valimehr
AU - D. Styczynski
AU - M. Mudaliyar
AU - S. S. Kundapura
AU - M. D. Johnson
AU - D. Ghosal
AU - E. Hanssen
PY - 2026
DA - 2026/09/27
PB - bioRxiv
DO - 10.64898/2026.09.27.753993
UR - https://www.biorxiv.org/content/10.64898/2026.09.27.753993v1
AB - The visualisation of large biological samples in cryo-electron tomography requires both few-nanometre resolution and a wide, ideally >10 micrometrre, field of view. For a single image, increasing magnification trades increased resolution for a smaller field of view. Montaging, where high-magnification images are stitched together to increase field of view, addresses the trade-off but is dose-inefficient with round beams that are standard in transmission electron microscopy (TEM), since this illumination shape does not tessellate. Our workflow greatly simplifies montaging schemes for dose-sensitive specimens, allowing large montages of any shape - from individual cells or other features of interest up to whole lamellae - to be collected at high resolution without compromising electron dose efficiency. We demonstrate the workflow by acquiring montage tomograms of whole yeast (Saccharomyces cerevisiae) cells in cryo-FIB lamellae and of human neurons.
ER -