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Correlative multiscale microCT-SBF-SEM imaging of resin-embedded tissue

Correlative multiscale microCT-SBF-SEM imaging of resin-embedded tissue
Correlative multiscale microCT-SBF-SEM imaging of resin-embedded tissue

Three-dimensional biological microscopy presents a trade-off between spatial resolution and field of view. Correlative approaches applying multiple imaging techniques to the same sample can therefore mitigate against these trade-offs. Here, we present a workflow for correlative microscopic X-ray microfocus computed tomography (microCT) and serial block face scanning electron microscopy (SBF-SEM) imaging of resin-embedded tissue, using mammalian placental tissue samples as an example. This correlative X-ray and electron microscopy (CXEM) workflow allows users to image the same sample at multiple resolutions, and target the region of interest (ROI) for SBF-SEM based on microCT. We detail the protocols associated with this workflow and demonstrate its application in multiscale imaging of horse placental villi and ROI selection in the labyrinthine zone of a mouse placenta. These examples demonstrate how the protocol may need to be adapted for tissues with different densities.

Correlative microscopy, CXEM, microCT, Multiscale, Placenta, SBF-SEM
0091-679X
241-267
Academic Press
Laundon, Davis
b2ba5687-d949-47dd-b84f-fb3c0c032550
Katsamenis, Orestis L.
8553e7c3-d860-4b7a-a883-abf6c0c4b438
Thompson, James
a0a1e940-d720-47de-81d7-ebcd48738239
Goggin, Patricia
e1e6172b-6e76-47aa-a2a6-3f2a7f4a0a7d
Chatelet, David S.
6371fd7a-e274-4738-9ccb-3dd4dab32928
Chavatte-Palmer, Pascale
3b1ff263-828e-4f55-ab66-13168e3ed45f
Gostling, Neil J.
4840aa40-cb6c-4112-a0b9-694a869523fc
Lewis, Rohan M.
caaeb97d-ea69-4f7b-8adb-5fa25e2d3502
Narayan, Kedar
Collinson, Lucy
Verkade, Paul
Laundon, Davis
b2ba5687-d949-47dd-b84f-fb3c0c032550
Katsamenis, Orestis L.
8553e7c3-d860-4b7a-a883-abf6c0c4b438
Thompson, James
a0a1e940-d720-47de-81d7-ebcd48738239
Goggin, Patricia
e1e6172b-6e76-47aa-a2a6-3f2a7f4a0a7d
Chatelet, David S.
6371fd7a-e274-4738-9ccb-3dd4dab32928
Chavatte-Palmer, Pascale
3b1ff263-828e-4f55-ab66-13168e3ed45f
Gostling, Neil J.
4840aa40-cb6c-4112-a0b9-694a869523fc
Lewis, Rohan M.
caaeb97d-ea69-4f7b-8adb-5fa25e2d3502
Narayan, Kedar
Collinson, Lucy
Verkade, Paul

Laundon, Davis, Katsamenis, Orestis L., Thompson, James, Goggin, Patricia, Chatelet, David S., Chavatte-Palmer, Pascale, Gostling, Neil J. and Lewis, Rohan M. (2023) Correlative multiscale microCT-SBF-SEM imaging of resin-embedded tissue. In, Narayan, Kedar, Collinson, Lucy and Verkade, Paul (eds.) Methods in Cell Biology: Volume Electron Microscopy. (Methods in Cell Biology, 177) Academic Press, pp. 241-267. (doi:10.1016/bs.mcb.2023.01.014).

Record type: Book Section

Abstract

Three-dimensional biological microscopy presents a trade-off between spatial resolution and field of view. Correlative approaches applying multiple imaging techniques to the same sample can therefore mitigate against these trade-offs. Here, we present a workflow for correlative microscopic X-ray microfocus computed tomography (microCT) and serial block face scanning electron microscopy (SBF-SEM) imaging of resin-embedded tissue, using mammalian placental tissue samples as an example. This correlative X-ray and electron microscopy (CXEM) workflow allows users to image the same sample at multiple resolutions, and target the region of interest (ROI) for SBF-SEM based on microCT. We detail the protocols associated with this workflow and demonstrate its application in multiscale imaging of horse placental villi and ROI selection in the labyrinthine zone of a mouse placenta. These examples demonstrate how the protocol may need to be adapted for tissues with different densities.

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More information

e-pub ahead of print date: 10 April 2023
Published date: 12 July 2023
Additional Information: Funding Information: This project was funded by Leverhulme grant number RPG-2019-208. Equipment in the Biomedical Imaging Unit was supported by MR/L012626/1 Southampton Imaging under MRC UKRMP Funding. Beamtime allocation at the μ-VIS X-ray Imaging Centre/3D X-ray Histology facility ( www.muvis.org/www.xrayhistology.org ) was supported by the National Research Facility for Lab-based X-ray CT (nxct.ac.uk) through EPSRC grant EP/T02593X/1.
Keywords: Correlative microscopy, CXEM, microCT, Multiscale, Placenta, SBF-SEM

Identifiers

Local EPrints ID: 484213
URI: http://eprints.soton.ac.uk/id/eprint/484213
ISSN: 0091-679X
PURE UUID: 2fbbbe81-02ff-47cc-896b-cef67686188e
ORCID for Davis Laundon: ORCID iD orcid.org/0000-0002-1508-664X
ORCID for Orestis L. Katsamenis: ORCID iD orcid.org/0000-0003-4367-4147
ORCID for James Thompson: ORCID iD orcid.org/0000-0002-9285-1317
ORCID for Neil J. Gostling: ORCID iD orcid.org/0000-0002-5960-7769
ORCID for Rohan M. Lewis: ORCID iD orcid.org/0000-0003-4044-9104

Catalogue record

Date deposited: 13 Nov 2023 18:40
Last modified: 20 Jan 2024 03:15

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Contributors

Author: Davis Laundon ORCID iD
Author: James Thompson ORCID iD
Author: Patricia Goggin
Author: David S. Chatelet
Author: Pascale Chavatte-Palmer
Author: Rohan M. Lewis ORCID iD
Editor: Kedar Narayan
Editor: Lucy Collinson
Editor: Paul Verkade

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