Multitime point transport of Aβ in the optic nerve. (A) Timeline of intracisternal injection of Aβ tracer. (B) Bioluminescence imaging showing the fluorescence intensity of Aβ in the brain and optic nerve 30 and 60 min after injection. (C) Representative images of Aβ distribution along the optic nerve in sections optic nerve after normal saline injection (30 min) and Aβ injection (30 min, 60 min). (D) Statistical graph of Aβ transportation distance in the optic nerve 30 and 60 min after injection (n = 5). (E) Representative images of short-range tracking of retinal Aβ transportation. (F) Fluorescence intensity profiles of the location of Aβ and IB4 in the vessels after a 30-min injection of CSF tracer (n = 8). (G) Statistical graphs of the percentage of area covered in short-range tracking experiments (n = 8). (H) Representative images showing the distribution of Aβ along the deep cervical lymph nodes (dcLNs) 30 min after saline injection and 30 min after Aβ injection. (I and J) Representative images and fluorescence intensity profiles of Aβ in the common carotid artery (CCA) and retina after normal saline injection (30 min) and Aβ injection (30 min) (n = 5). (K) Representative images and fluorescence intensity statistics showing the distribution of Aβ along the retina 30 min after Aβ injection following optic nerve ligation. (L) Immunofluorescence staining images of Aβ, Lyve1, CD31, and CD45 at the periorbital lymphatics, with CD45 labeling macrophages to exclude the possibility that Lyve1-labeled lymphatics are macrophages. Data are representative of three independent experiments. All data are presented as mean ± SEM. Statistical analysis was performed using two-tailed unpaired t tests (D, G, and K) or two-way ANOVA with post hoc Tukey tests (J).