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GCIB-SEM: A path to 10 nm isotropic imaging of cubic millimeter volumes

K.J. Hayworth, D. Peale, M. Januszewski, G.W. Knott, Z. Lu, C.S. Xu, H.F. Hess
doi: https://doi.org/10.1101/563239
K.J. Hayworth
HHMI Janelia Research Campus
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D. Peale
HHMI Janelia Research Campus
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M. Januszewski
Google
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G.W. Knott
EPFL
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Z. Lu
Dalhousie University
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C.S. Xu
HHMI Janelia Research Campus
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H.F. Hess
HHMI Janelia Research Campus
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Abstract

Focused Ion Beam Scanning Electron Microscopy (FIB-SEM) generates 3D datasets optimally suited for segmentation of cell ultrastructure and automated connectome tracing but is limited to small fields of view and is therefore incompatible with the new generation of ultrafast multibeam SEMs. In contrast, section-based techniques are multibeam-compatible but are limited in z-resolution making automatic segmentation of cellular ultrastructure difficult. Here we demonstrate a novel 3D electron microscopy technique, Gas Cluster Ion Beam SEM (GCIB-SEM), in which top-down, wide-area ion milling is performed on a series of thick sections, acquiring < 10 nm isotropic datasets of each which are then stitched together to span the full sectioned volume. Based on our results, incorporating GCIB-SEM into existing single beam and multibeam SEM workflows should be straightforward and should dramatically increase reliability while simultaneously improving z-resolution by a factor of 3 or more.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY 4.0 International license.
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Posted February 28, 2019.
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GCIB-SEM: A path to 10 nm isotropic imaging of cubic millimeter volumes
K.J. Hayworth, D. Peale, M. Januszewski, G.W. Knott, Z. Lu, C.S. Xu, H.F. Hess
bioRxiv 563239; doi: https://doi.org/10.1101/563239
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GCIB-SEM: A path to 10 nm isotropic imaging of cubic millimeter volumes
K.J. Hayworth, D. Peale, M. Januszewski, G.W. Knott, Z. Lu, C.S. Xu, H.F. Hess
bioRxiv 563239; doi: https://doi.org/10.1101/563239

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