Electron tomography of microtubule end-morphologies in C. elegans embryos.
In this chapter we describe the preparation of early mitotic C. elegans embryos for the tomographic reconstruction of end-morphologies of spindle microtubules. Early embryos are prepared by high-pressure freezing and freeze-substitution for thin-layer embedding in Epon/Araldite. We further describe data acquisition, tomographic reconstruction, and 3-D modeling of microtubules in serially sectioned mitotic spindles. The presented techniques are applicable to other model systems.
O'Toole
Eileen
E
The Boulder Laboratory for 3-D Electron Microscopy of Cells, University of Colorado, Boulder, CO, USA.
Müller-Reichert
Thomas
T
D000818:Animals; D017173:Caenorhabditis elegans / Q000196:embryology*; D055032:Electron Microscope Tomography / Q000379:methods; D004625:Embryo, Nonmammalian / Q000648:ultrastructure; D017110:Freeze Substitution / Q000379:methods; D021621:Imaging, Three-Dimensional; D008870:Microtubules / Q000648:ultrastructure*