Fig. 4

Evaluation of glymphatic system function in the ischemia–reperfusion stroke model using NIR-II fluorescence imaging. a Workflow of the in vivo imaging experiment. The schematic diagram was created by using the online software https://biorender.com. b Representative NIR-II images showing impaired glymphatic system function after MCAO and undamaged glymphatic function in sham mice. Time-series of fluorescence signal variation in each hemisphere of MCAO mice c and sham mice d. White scale bar: 2 mm. Comparison of mean intensity between contralateral hemisphere and ipsilateral hemisphere at 10 min e, two-way ANOVA with Sidak’s multiple comparisons test, MCAO: P = 0.1758 < Sham: P = 0.9727 and 30 min f, two-way ANOVA with Sidak’s multiple comparisons test, MCAO: P = 0.1658 < Sham: P = 0.8214, n = 4 for MCAO group, n = 5 for sham group. g Schematic illustrating the quantification process of the ratio. The schematic diagram was created by using the online software https://biorender.com. h Time-series of ratio within 30 min after CM infusion initiation in MCAO mice and sham mice, n = 5 per group, two-way ANOVA with Sidak’s multiple comparisons test, 5 min: P = 0.0085, 10 min: P < 0.0001, 15 min: P = 0.0005, 20 min: P = 0.0002, 25 min: P = 0.0012, 30 min: P = 0.0037. i The NIR-II tracer filling rate in MCA region in MCAO mice and sham mice calculated from the intensity variation within 30 min duration, n = 4 for MCAO group, n = 5 for sham group, two-way ANOVA with Sidak’s multiple comparisons test, MCAO: P = 0.0219, Sham: P = 0.5736. j Imaging of the NIR-II tracer distribution on the brain surface at the dorsal and ventral positions. White scale bar: 2 mm. The ratio of MCAO mice and sham mice at the dorsal k and ventral l positions, n = 5 per group, t test, Dorsal: P = 0.0336, Ventral: P = 0.1951, Colored dot represents the ratio of individual animal. Center is median and quartiles shown by box and whiskers