Extended Data Fig. 13: Expansion workflow and imaging results for C. elegans embryos, related to Fig. 3d.

a) Immobilization, permeabilization, fixation, immunostaining, and expansion takes approximately four days. b) Overlay of embryo stained with DAPI before and after expansion after 12-degree affine registration (cyan: pre-expansion; magenta: post-expansion). The embryo was imaged with diSPIM. Single-view maximum intensity projection along xy and xz views are shown for comparison. The registration results from 7 embryos (normalized cross-correlation: 0.72 ± 0.03) show that the expansion factor for whole embryos is 3.29 ± 0.14, with nearly isotropic expansion (3.24 ± 0.14, 3.39 ± 0.14, 3.26 ± 0.17 for x, y and z dimensions, respectively). Scale bar: 5 µm in pre-expansion units. c) Magnified view of the rectangle shown in b), showing no obvious local distortions in nuclei shape during expansion. Scale bar: 2 µm in pre-expansion units.