Visudyne

Mice were anesthetized via i.p. ketamine (100 mg/mL)/xylazine (10 mg/mL). Visudyne (Valeant Ophtalmics) was reconstituted following manufacturer instructions and 5 μL was injected i.c.m. After 15 minutes, Visudyne was photoconverted with a nonthermal 689-nm wavelength laser (Coherent Opal Photoactivator, Lumenis) on 5 different spots through the intact skull (1 on the cisterna magna, 2 on the transverse sinuses, 1 on the superior sagittal sinus, and 1 at the confluence of sinuses). Each spot was irradiated with a light dose of 50 J/cm2 at an intensity of 600 mW/cm2 for 83 s. Mice were sutured and allowed to recover on a heating pad followed by postsurgical analgesic administration. Mice were utilized for experiments seven days after lymphatic ablation. Functional parameters following Visudyne treatment and lack of potential off target effects have been thoroughly characterized using an identical paradigm (Da Mesquita et al., 2018 (link); Louveau et al., 2018 (link)).

Adapted from previous publications^{28 (link),51 (link),52 (link)}. Mice were anesthetized with ketamine/xylazine (i.p.). Visudyne® (Valeant Ophtalmics) was reconstituted following manufacturer instructions and 5µl was injected either i.c.m into the cisterna magna (for meningeal lymphatic ablation) or 2µl in each nostril (for nasal lymphatics ablation). After 15 min, Visudyne was photo-converted with a non-thermal 689nm wavelength laser light (Coherent Opal Photoactivator, Lumenis) on 5 different spots through the intact skull (1 on the cisterna magna, 2 on the transverse sinuses, 1 on the superior sagittal sinus and 1 at the junction of all sinuses). For nasal lymphatic ablation, the laser was applied through the soft palate of the mouse’s mouth. Each spot was irradiated with a light dose of 50J/cm² at an intensity of 600mW/cm² for 83s. Mice were then sutured and allowed to recover on a heating pad until responsive. Post-surgery, mice were given analgesic subcutaneously.