Direct comparison of syringeal soft tissue imaging techniques reveals advantages of μCT. (A) Frontal section of a male zebra finch syrinx prepared with conventional histology and stained with hematoxylin and eosin. Muscles appear as bright red, cartilage as speckled blue/purple. (B) Virtual frontal section through a 3D MRI dataset of a Magnevist-contrasted male zebra finch syrinx with isotropic voxel resolution of 23 μm. (C) Virtual frontal section through a 3D μCT dataset of an iodine-contrasted male zebra finch syrinx with an isotropic voxel resolution of 5 μm. The non-destructiveness, high resolution and relatively short scanning times of contrasted samples made μCT the optimal technique for the construction of the syrinx morphome. (D) Volume rendering of an iodine-contrasted μCT scan. (E-G) Virtual horizontal sections through the 3D μCT dataset at different positions as shown in D. Special effort was made to fix and scan the syrinx in situ, with surrounding tissues remaining intact. Abbreviations as listed in Table 1. Scale bars: 1 mm.