Light sheet volumetric image of the hypothalamus of an E20 chick embryo. An E20 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 rotation of a maximum intensity projection 3D render of the volume, dorsal is to the left in the first frame.