Protocol for volume correlative light X-ray and electron microscopy of endothelial cells in mouse tissue

Natalia Reglero-Real; Lorena Pérez-Gutiérrez; Rebeca S. Saleeb; Sussan Nourshargh; Lucy Collinson; Azumi Yoshimura

STAR Protocols (Sep 2024)

Protocol for volume correlative light X-ray and electron microscopy of endothelial cells in mouse tissue

Natalia Reglero-Real,
Lorena Pérez-Gutiérrez,
Rebeca S. Saleeb,
Sussan Nourshargh,
Lucy Collinson,
Azumi Yoshimura

Affiliations

Natalia Reglero-Real: Centre for Microvascular Research, William Harvey Research Institute, Barts and the London School of Medicine and Dentistry, Queen Mary University of London, EC1M 6BQ London, UK; Departamento e Instituto de Biología Molecular (IUBM, UAM), Centro de Biología Molecular Severo Ochoa, Universidad Autónoma de Madrid, UAM-CSIC, Campus de Cantoblanco, 28049 Madrid, Spain; Corresponding author
Lorena Pérez-Gutiérrez: Centre for Microvascular Research, William Harvey Research Institute, Barts and the London School of Medicine and Dentistry, Queen Mary University of London, EC1M 6BQ London, UK; Cancer Immunology and Immunotherapy Lab, Center for Cooperative Research in Biosciences (CIC BioGUNE), Basque Research and Technology Alliance (BRTA), Derio, 48160 Bizkaia, Spain
Rebeca S. Saleeb: Centre for Microvascular Research, William Harvey Research Institute, Barts and the London School of Medicine and Dentistry, Queen Mary University of London, EC1M 6BQ London, UK; Institute of Regeneration and Repair, University of Edinburgh, EH16 4UU Edinburgh, UK
Sussan Nourshargh: Centre for Microvascular Research, William Harvey Research Institute, Barts and the London School of Medicine and Dentistry, Queen Mary University of London, EC1M 6BQ London, UK
Lucy Collinson: Electron Microscopy Science Technology Platform, The Francis Crick Institute, NW1 1AT London, UK; Corresponding author
Azumi Yoshimura: Electron Microscopy Science Technology Platform, The Francis Crick Institute, NW1 1AT London, UK; Graduate School of Medicine, Yamaguchi University, Ube 755-8505, Japan; Corresponding author

Journal volume & issue: Vol. 5, no. 3
p. 103257

Abstract

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Summary: Correlative light and electron microscopy (CLEM) greatly facilitate capturing the ultrastructure of spatially and/or temporally rare events. Here, we present a protocol for targeting regions of interests (ROIs) in tissue endothelial cells (ECs) using X-ray micro-computed tomography (μCT). We describe steps for ROI targeting guided by vasculature patterns and positions of EC nuclei visualized by light and X-ray microscopy. The protocol is applicable to thin or translucent tissues that contain defined landmarks visible in both light and X-ray microscopy.For complete details on the use and execution of this protocol, please refer to Reglero-Real et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

Published in STAR Protocols

ISSN: 2666-1667 (Online)
Publisher: Elsevier
Country of publisher: United States
LCC subjects: Science: Science (General)
Website: https://star-protocols.cell.com/

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