Fig. 2.

Dual IFL for NPY (Alexa-647, imaged in red channel) and TH (Cy3, imaged in green channel) within the medial hypothalamus of a clarified intact (whole) brain hybrid prepared from a newborn rat pup (P0) perfused with Hydrogel solution A. a Maximum intensity flattened Z-projection obtained from 262 1.5 µm-thick optical frames (393 µm). This confocal image was obtained using a ventral approach. The total volume of the flattened image is 0.137 mm³ (i.e., 591 µm × 591 µm × 393 µm). The inset in a is a photograph of this hybrid sample, positioned ventral side up, that was obtained during confocal image collection. b Rotated view of the z-stack depicted in a (ventral is at the top, V). See the supplementary video for a 3D rendering of the entire volume. Scale bar in a 100 µm. 3V third ventricle; R rostral (color figure online)