Laboratory of Molecular and Genetic Cardiovascular Pathophysiology, Vascular Biology Area, Centro Nacional de Investigaciones Cardiovasculares (CNIC), Madrid, Spain.
Centro de Investigación Biomédica en Red de Enfermedades Cardiovasculares (CIBERCV), Madrid, Spain.
Methods Mol Biol. 2022;2419:597-610. doi: 10.1007/978-1-0716-1924-7_37.
Confocal imaging of the mouse aorta is a powerful, indispensable technique for the study of cardiovascular pathology ex vivo. Whole mount en face preparations allow visualization of wide areas of the luminal vessel surface, thus enabling a thorough analysis of multiple cellular and structural features of the endothelial cell-rich intimal layer. This method is a suitable tool for the study of endothelial cell dysfunction and leukocyte infiltration, both of which contribute to the onset of pathological vascular conditions such as atherosclerosis. This chapter provides a complete guide on how to perfuse-fix mouse aorta, dissect the vessel, immunostain target proteins, and carry out en face confocal image acquisition and analysis.
共聚焦成像技术是研究离体心血管病理学的强大、不可或缺的工具，可对小鼠主动脉进行共聚焦成像。全血管平铺明场制备可使管腔表面的大面积区域可视化，从而能够对富含内皮细胞的内膜层的多个细胞和结构特征进行全面分析。该方法是研究内皮细胞功能障碍和白细胞浸润的合适工具，这两者均会导致动脉粥样硬化等病理血管状况的发生。本章提供了一个完整的指南，介绍如何对小鼠主动脉进行灌流固定、血管解剖、免疫染色靶蛋白，以及进行明场共聚焦图像采集和分析。
