To analyze the overall three-dimensional shape changes during H/H and H/P fold formation over time, we developed a protocol for live imaging of wing imaginal discs in culture (Methods). Cultured wing imaginal discs sustained cell proliferation for at least 10 h (Supplementary Fig. 1m, Supplementary Movie 1) and formed H/H and H/P folds with no visible difference in shape from the hinge folds of fixed wing imaginal discs (Supplementary Figure 1n–q, Supplementary Movie 1).