Description of study: To characterize the effect of Chiron on the shape transition, we analyzed samples treated with Chiron as well as samples without Chiron, at day 2, 3 and 4 after aggregation (3 of each: 15 samples in total). To make the nuclei visible, they were stained with Sytox DeepRed and imaged in far red light. The samples treated with Chiron were also imaged in the red and green channel to visualize the mesoderm and endoderm germ layers. Figure 1 shows typical gastruloid shapes imaged in 2D brightfield microscopy.
Figure 1. Brightfield images of mouse gastruloids at 48, 72, 96 and 120h (2,3,4, 5 days) after aggregation (2,3,4, 5 days). Chiron pulse between 48 and 96h. Credit: Sergei Ponomartcev.
Figure 2. 2D Fluorescence images of mouse gastruloids at 48, 72, 96 and 120 h (2,3,4 and 5 days after aggregtion, dpa). Chiron pulse between 48 and 96h. Credit: Sergei Ponomartcev. Size of 3-day gastruloid ~300 um.
