Adult Prox1GFP, LckCre::tdTOMATOfl/fl mice were anesthetized by ketamine/xylazine intraperitoneal (i.p.) injection. Mice were injected intravenously (i.v.) or into the cisterna magna (i.c.m.) with Qtracker655 vascular tracker (ThermoFisher - Qdot655 – 5µl with 95µl saline (i.v.) or 5µl (i.c.m.)). Furamidine dihydrochloride (125µg -Sigma Aldrich) was injected i.v. prior to imaging. Meningeal lymphatic vessels were imaged through a thinned skull preparation above the superior sagittal, transverse sinuses or cerebellum. The core temperature of the mice was maintained at 37C. Imaging was performed with a Leica TCS SP8 multiphoton system (Leica Microsystems) equipped with a Chameleon Ultra II tunable Ti:sapphire laser (Coherent). GFP, tdTOMATO and Qdot655 were excited with an excitation wavelength of 880 nm (or 780nm for furamidine57 (link)). Second harmonic signal (bone and meningeal signal) was visualized using a 400/450 external detector. Images were obtained using a 25X water immersion objective with 0/95NA and external HyD non-descanned detectors (Leica Microsystems). Three-dimensional images data were collected by obtaining images from the x,y and z-planes. The resulting images were analyzed with Imaris software (Bitplane).