(A) Z-projection of the PLLp at ~32 hpf from a single deconvolved DiSPIM volume. Red and blue dashed lines indicate the position of cross-sections in B and C, respectively. Membranes are indicated in green and nuclei in magenta. (B) Length-wise section of the PLLp at the position indicated in by the red line in A. Arrowheads indicate superficial nuclei. (C) Cross-section of the PLLp at the position indicated by the blue line in A. Arrowheads indicate superficial nuclei. (D) Single processed Airyscan super-resolution confocal slice adjacent to the skin showing superficial nuclei. (E) Schematic of nuclear position in a single PLLp taken from an Airyscan-processed confocal stack. Gray dots indicate all PLLp nuclei, red dots indicate the position of superficial nuclei (see text for details), and blue dots indicate the position of protoneuromast apical constrictions. (F) Average of the positions of nuclei in n = 10 PLLp taken from Airyscan confocal stacks. PLLp were aligned based on the position of the leading-most protoneuromast. Gray indicates the position of all nuclei, red filled topo-lines indicate the position of superficial nuclei and blue open topo-lines indicate the position of protoneuromast apical constrictions. X and Y axes in E, F are distance in microns from the center of the first formed protoneuromast, located at 0,0. E,F are top view of the PLLp as in panel G. (G,H, I) 3D reconstruction of a DiSPIM volume of a ~32 hpf PLLp showing the superficial cells in magenta and the remaining cells in green in top (G), bottom (H), and side (I) views.