Abstract
This chapter presents a summary of standard structural and immunocytochemical methods considered as the most basic for any ultrastructural study. Three-dimensional (3D) imaging techniques in electron microscopy (EM) have been greatly improved in two areas: (1) electron tomography of cell organelles or cell sections and (2) reconstruction of macromolecules from single particles. X-ray crystallography has been a highly useful technique to elucidate the structure of large individual proteins. However, X-ray crystallography becomes limited when the structure of multicomponent complexes needs to be analyzed. Cryo-EM is a powerful tool to investigate the structure, assembly, conformational changes and dynamic interactions of biological supramolecules, and provides unique information for understanding large, complex biological systems. Electron tomography of frozen-hydrated tissue sections enables the analysis of the 3D structure of the specimen in situ , keeping the cells in near-native state. Cryo-tomography has the potential of visualizing the 3D organization of cells and organelles at molecular resolution and it is a non-invasive method.
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