Experimental observation of gastruloids and shape quantification using LOCO-EFA.

A: Representative images of fixed gastruloids after 48 h, 72 h and 96 h. Cell nuclei were stained with DAPI. Shown is the mid-plane of a z-stack. Scale bar: 100 μm (48 h and 72 h), 200 μm (96 h). B: Image of a gastruloid with a binary mask created from that image. Numerical values of L2/L1 and L3/L1 for this gastruloid are given. Scale bar: 100 μm. C: LOCO-EFA analysis of measured gastruloids at 72 h (n = 132, two biological replicates) and 96 h (n = 413, 8 biological replicates). Each data point is a gastruloid. The scatter plot shows the scaled LOCO-EFA coefficients L2/L1 and L3/L1. The 3 insets show images of the gastruloids highlighted in the LOCO-EFA scatter plot. Cell nuclei were stained with DAPI. Shown is the mid-plane of a z-stack. For each gastruloid, the scaled LOCO-EFA coefficients L2/L1 and L3/L1 are given. Scale bar: 100 μm.