Light sheet volumetric image of the hypothalamus of an E16 chick embryo. An E16 embryo was perfused, fixed with 4% PFA, and the brain was dissected out. The isolated brain was CUBIC-cleared, and immunostained with anti-orexin (red), anti-TH (white) and anti-c-Fos (green) antibodies. Nuclei were counterstained with DAPI (blue). Coronal images were acquired every 5μm using a 5x objective. The movie is a coronal stack animation from dorsal to ventral; the third ventricle runs diagonally from the upper left-hand-side corner to the bottom-center.