Fig. 3.

Time series of zebrafish craniofacial development. (A) Zebrafish larvae were analyzed from 2 to 6 dpf. Scanning electron microscope (SEM) images confirmed the soft tissue morphology captured by DAPI staining and confocal microscopy used in zFACE. (B) Changes in zFACE features were followed across developmental time points, and alterations in 28 measurements were determined (representative changes are shown in the graphs). The numbers of individual larvae for each group were as follows: n=30 at 3 dpf, n=31 at 4 dpf, n=28 at 5 dpf and n=28 at 6 dpf. Specifically, 20 zFACE measurements changed between 3 and 4 dpf, 12 changed between 4 and 5 dpf, and 6 nominally changed between 5 and 6 dpf. *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001; n.s., not significant (one-way ANOVA). (C) Untransformed zFACE features were subjected to multivariate statistical testing using principal component analysis (PCA). The first two components captured 66% of the variance in the dataset. There were differences in developmental times that were captured across PC1, with no strain-specific differences observed. Scale bars: 50 µm.