Three-dimensional reconstruction methods for Caenorhabditis elegans ultrastructure

Abstract The roundworm Caenorhabditis elegans is one of the major model organisms in modern cell and developmental biology. Here, we present methods for the three-dimensional (3D) reconstruction of the worm ultrastructure. We describe the use of (1) serial-section analysis, (2) electron tomography, and (3) serial block face imaging by scanning electron microscopy (SEM). Sample preparation for high-pressure freezing/freeze substitution (HPF/FS) has been extensively covered in a previous volume of this “Methods in Cell Biology” series and will only be described briefly. We will discuss these 3D methods in light of recent research activities related to worm and early embryo biology.