High resolution imaging of the cortex isolated from sea urchin eggs and embryos

• Published in: Methods in cell biology Vol. 151; pp. 419 - 432
• Main Authors: Henson, J H, Samasa, Bakary, Burg, E C
• Format: Journal Article
• Language: English
• Published: United States 2019
• Subjects: Animals; Cell Membrane - ultrastructure; Cytoplasm - ultrastructure; Cytoskeleton - ultrastructure; Embryo, Nonmammalian; Exocytosis - genetics; Fertilization - genetics; Molecular Imaging - methods; Ovum - ultrastructure; Sea Urchins - growth & development; Sea Urchins - ultrastructure; Embryo; Egg; Fertilization; Cortex; Cytoskeleton; Cytokinesis; Sea urchin
• ISSN: 0091-679X
• DOI: 10.1016/bs.mcb.2019.01.005

The cellular cortex-consisting of the plasma membrane and the adjacent outer few microns of the cytoplasm-is a critically important, dynamic and complex region in the sea urchin egg and embryo. Some 40 years ago it was discovered that isolated cortices could be obtained from eggs adhered to glass coverslips and since that time this preparation has been used in a wide range of studies, including seminal research on fertilization, exocytosis, the cytoskeleton, and cytokinesis. In this chapter, we discuss methods for isolating cortices from eggs and embryos, including those undergoing cell division. We also provide protocols for analyzing cortical architecture and dynamics using specific localization methods combined with super-resolution Structured Illumination and Stimulated Emission Depletion light microscopy and platinum replica transmission electron microscopy.