3D Reconstruction of Chicken Embryo Development

Fertilized White Leghorn chicken (Gallus gallus) eggs were incubated in a humidified atmosphere at 37.0°C until HH11–13 [7] (link). The embryos were isolated (HH11 n = 6; HH12 n = 10; HH13 n = 13), fixed overnight in 4% paraformaldehyde, included in paraffin, sectioned and stained for hematoxylin and eosin (H&E) as previously described [18] (link). Sections were imaged on an Olympus AX70 microscope (Olympus) equipped with an Olympus XC50 camera (Olympus). For 3D reconstructions, serial paraffin transversal sections of freshly isolated embryos at stage HH12 and HH13 stained with H&E were digitalized using a Panoramic MIDI scanner (3D Histech) and reconstructed with Amira 4.1 software (Visage Imaging).