Ultrastructure Expansion Microscopy applied to C. elegans embryos

Visualization of organelles using expansion microscopy has been previously applied to Caenorhadbitis elegans adult gonads or worms. However, its application to embryos has remained a challenge due to the protective eggshell barrier. Here, by combining freeze-cracking and ultrastructure expansion microscopy (U-ExM), we demonstrate a four-time isotropic expansion of C. elegans embryos. As an example structure, we chose the nuclear pore and demonstrate that we achieve sufficient resolution to distinguish them individually. Our work provides proof of principle for U-ExM in C. elegans embryos, which will be applicable for imaging a wide range of cellular structures in this model system.