Fig. 4.

Generation of forebrain-patterned organoids from mouse EpiSCs. (A) Diagram of the protocol for forebrain-patterned neural organoid generation. (B) Brightfield images of forebrain organoids. (C) Confocal immunofluorescence image of a developing organoid after 48 h (d2). (D,E) Confocal immunofluorescence images of a d4 organoid. (F) z-stack reconstruction of a d4 organoid. (G) Confocal immunofluorescence image of a day 8 organoid. Scale bars: 200 μm (B); 25 μm (C-G). For C-G, all organoids were cleared (see Materials and Methods) and imaged in toto without sectioning.