and systems have the potential to provide a framework to study the organization, gene expression, and functionality of lymphatic and blood vessel smooth muscle cells in physiology and disease settings. A series of procedures are described here, including the surgical isolation of mouse collecting lymphatic vessels and blood vessels, whole-mount immunofluorescence staining of muscle cells on the vessels, and the enzymatic digestion and culture of α- smooth muscle actin+ cells from the vessels. For complete details on the use and execution of this protocol, please refer to Jones et al. (2018).