Experimental steering of electron microscopy studies using prior X-ray computed tomography

•Using microCT pre-scans to accurately steer serial block face SEM.•High throughput screening and mapping samples to reduce time hunting for features of interest.•Using microCT to optimise specimen preparation and staining.•Using microCT to guide site-specific TEM sample preparation. Transmission el...

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Veröffentlicht in:Ultramicroscopy 2019-06, Vol.201, p.58-67
Hauptverfasser: Starborg, Tobias, O'Sullivan, James D.B., Carneiro, Claudia Martins, Behnsen, Julia, Else, Kathryn J., Grencis, Richard K., Withers, Philip J.
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Sprache:eng
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Zusammenfassung:•Using microCT pre-scans to accurately steer serial block face SEM.•High throughput screening and mapping samples to reduce time hunting for features of interest.•Using microCT to optimise specimen preparation and staining.•Using microCT to guide site-specific TEM sample preparation. Transmission electron microscopy (TEM) and scanning electron microscopy (SEM) can provide unrivalled high-resolution images of specific features and volumes of interest. However, the regions interrogated are typically very small, and sample preparation is both time-consuming and destructive. Here we consider how prior X-ray micro-computed tomography (microCT) presents an opportunity to increase the efficiency of electron microscopy in biology. We demonstrate how it can be used to; select the most promising samples and target site-specific locations; provide a wider context of the location being interrogated (multiscale correlative imaging); guide sample preparation and 3D imaging schemes; as well as quantify the effects of destructive sample preparation and staining procedures. We present a workflow utilising open source software in which microCT can be used either broadly, or precisely, to experimentally steer and inform subsequent electron microscopy studies. As automated sample registration procedures are developed to enable correlative microscopy, experimental steering by prior CT could be beneficially routinely incorporated into many experimental workflows.
ISSN:0304-3991
1879-2723
DOI:10.1016/j.ultramic.2019.03.002