Fig. 3
Light sheet imaging of the whole mouse brain and of connections at the skull-brain interface visualized using endogenous fluorophores preserved by ADAPT-3D. (A) 3D whole-mount projection of the brain from a ChATCre;tdTomatofl/fl mouse (white) injected retro-orbitally with Lectin-Dylight649 to label vasculature (fire) acquired by light sheet microscopy. (B) Extended display near lateral ventricle from (A) displaying blood vessels in the core of the brain and preservation of fine neuron dendrites in the cortex. (C) 3D whole-mount projection of a brain from a 16-week old mouse expressing CD11c-eYFP which was decolorized, delipidated, and incubated in ADAPT:RI followed by imaging with light sheet microscopy. (D) 500-micron coronal maximum intensity projection from whole brain of CD11c-eYFP where white arrow points to a CD11c-positive neuron. (E) A 50-micron x-y maximum intensity projection of the brain borders from a light sheet image of the whole skull from a Lyve1CreER;tdTomatofl/fl (red) mouse injected i.v. with Lectin-Dylight649 and CD31-AF647 to label blood vessels (white). The layers of the brain borders are annotated with the following abbreviations: muscle [MS], skull [SK], dura mater [D], leptomeninges [LM], and brain parenchyma [B]. (F) Dorsal view of the light sheet imaging volume acquired from the whole skull in E where arrowheads point to dural lymphatics and full arrows point to meningeal macrophages. (G) Extended display of Lyve1 positive skull channels where the asterisk denotes consecutive skull channels bridging the skull bone marrow and meninges. (H) A graphical depiction of the relationship between the depth of light penetration in light sheet images of intact mouse skulls and the corresponding tissue preparation times reported for the different clearing protocols in the literature including the 8-day time frame determined here for ADAPT-3D.
