Electron microscopy, immunostaining, cytoskeleton visualization, in situ hybridization, and three-dimensional reconstruction of Xenopus oocytes

Abstract

Although the overwhelming development of molecular techniques in recent decades has made ultrastructural studies less popular, to the point that ultrastructural interpretation is becoming a dying art, it still remains an indispensable tool for cell and developmental biologists. The introduction of EM-immunocytochemistry and three-dimensional visualization methods allows us to complement the knowledge gained from ultrastructural and molecular approaches. Because the first clues about the functions of newly discovered genes often come from the subcellular localization patterns of their proteins or RNAs, in this chapter we describe the methods that allow for precise ultrastructural localization and visualization of protein and RNA molecules within the compartments, organelles, and cytoskeleton of Xenopus oocytes.