Extended Data Fig. 8: rCATS across the adult mouse brain.

a, Serial coronal sections of a brain from a perfusion fixed adult mouse, revealing organ architecture with rCATS (WGA-CF633) in confocal overview imaging. Light areas typically correspond to regions rich in cell bodies or fiber tracts. Data are acquired across serial sections from the same biological specimen. b, rCATS in various example regions at two different magnifications, using WGA-AF594 or WGA-CF633 (see Supplementary Table 1 for detailed parameters). (Left) confocal, (right) xy-STED revealing differential tissue organization in different brain regions. The overview in the somatosensory cortex covers the superficial molecular layer (layer I, upper right corner) and layers II/III below. In the thalamus, imaging was performed in the ventral posterior nucleus, with the ad-axonal line in many of the fibers suggestive of myelination. In the medulla, bundles of thick nerve fibers are prominent. The ad-axonal line points to a myelination of these fibers. Imaging of the cerebellar cortex was performed in the vermal region, with the granular, Purkinje, and molecular layers visible in the overview. STED imaging was performed in the granular layer. In the hippocampal CA3 region overview, the different layers are conspicuous (left to right: stratum radiatum, stratum lucidum, stratum pyramidale). In the overview of the DG (left), the hilar region in the center is bordered by the granule cell layer above and below. The STED image (right) shows the transition between granule cell layer (top) and molecular layer (bottom) of the lower DG blade. In the CA1 region, the following layers can be discerned in the overview from top to bottom: stratum oriens, stratum pyramidale, stratum radiatum, stratum lacunosum moleculare. The STED image is from the transition between stratum pyramidale and stratum radiatum. All images are raw data. Imaging data in b were acquired across brain slices from n = 6 biological specimens.