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Updated: Jun 14, 2025
Retrograde Perfusion and Filling of Mouse Coronary Vasculature as Preparation for Micro Computed Tomography Imaging
Published on: February 10, 2012
Protocol for volume correlative light X-ray and electron microscopy of endothelial cells in mouse tissue
Natalia Reglero-Real1, Lorena Pérez-Gutiérrez2, Rebeca S Saleeb3
1Centre 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.
Abstract:
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 (microCT). 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.
