Intravital imaging of lymphatic vessels in lungs of ventilated, anesthetized mice. (A) 3D maximum intensity projections of the cleared lung from the Prox1-eGFP mouse showing paucity of lymphatics near distal pleural surfaces and prominent collecting lymphatics surrounding the pulmonary vein. (B) Distal lung intravital microscopy imaging showing initial lymphatic in Prox1-eGFP:Rosa26^mTmG mouse. (C) Surgical preparation, window design, and placement of window for imaging around the pulmonary vein on the mediastinal pleural surface. (D and E) Intravital imaging of functional collecting lymphatics in lungs of (D) Prox1-eGFP:Rosa26^mTmG and (E) Rosa26^mTmG mice. All images are representative of three or more independent experiments.