Fig. 4

Visualising internal structures of hCSs using Airy-beam light sheet fluorescent microscopy. a Clearvolume 3D representation of multiple internalised rosette lumen of differing sizes shown by RDYPROBE staining (F-actin) with dimensional scale. b Magnified region (a, dotted line) showing characteristic ovoid or tubular shaped lumens expected of a ventricle-like structure. c Composite (i) and individual channel max-projected images of (ii) RDYPROBE (F-actin) staining and (iii) Nestin-positive filaments around apical membranes, showing the presence of multiple large rosettes, Scale bars = 100 μm. d 3D render of magnified region from ci (dotted line) displaying of a singular rosette lumen co-stained for radial Nestin filaments and RDYPROBE (F-actin) with dimensional scale. e Orthogonal views of stained tissues permit examination of a cortical rosette in all axes. Scale bar = 50 μm