Pittsburgh Heart, Lung and Blood Vascular Medicine Institute, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania.
Division of Hematology/Oncology, Department of Medicine, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania.
Curr Protoc Cytom. 2020 Dec;95(1):e80. doi: 10.1002/cpcy.80.
Live imaging is critical to determining the dynamics and spatial interactions of cells within the tissue environment. In the lung, this has proven to be difficult due to the motion brought about by ventilation and cardiac contractions. A previous version of this Current Protocols in Cytometry article reported protocols for imaging ex vivo live lung slices and the intact mouse lung. Here, we update those protocols by adding new methodologies, new approaches for quantitative image analysis, and new areas of potential application. © 2020 Wiley Periodicals LLC. Basic Protocol 1: Live imaging of lung slices Support Protocol 1: Staining lung sections with fluorescent antibodies Basic Protocol 2: Live imaging in the mouse lung Support Protocol 2: Intratracheal instillations Support Protocol 3: Intravascular instillations Support Protocol 4: Monitoring vital signs of the mouse during live lung imaging Support Protocol 5: Antibodies Support Protocol 6: Fluorescent reporter mice Basic Protocol 3: Quantification of neutrophil-platelet aggregation in pulmonary vasculature Basic Protocol 4: Quantification of platelet-dependent pulmonary thrombosis Basic Protocol 5: Quantification of pulmonary vascular permeability.
实时成像对于确定组织环境中细胞的动力学和空间相互作用至关重要。在肺部，由于通气和心脏收缩引起的运动，这已被证明是困难的。本《当代细胞分析方法》文章的前一个版本报告了用于对离体活肺切片和完整的小鼠肺进行成像的方案。在这里，我们通过添加新的方法、新的定量图像分析方法和新的潜在应用领域来更新这些方案。© 2020 威利父子公司。基本方案 1：肺切片的实时成像支持方案 1：用荧光抗体对肺切片进行染色基本方案 2：在小鼠肺部进行实时成像支持方案 2：气管内滴注支持方案 3：血管内滴注支持方案 4：在活体肺成像期间监测小鼠的生命体征支持方案 5：抗体支持方案 6：荧光报告小鼠基本方案 3：肺血管中中性粒细胞-血小板聚集的定量基本方案 4：血小板依赖性肺血栓形成的定量基本方案 5：肺血管通透性的定量。
