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- W2505111835 abstract "This chapter explains the formation of a three-dimensional (3D) image of a macromolecule in its entirety, at the highest possible resolution. Specimen preparation methods perform the seemingly impossible task of stabilizing the initially hydrated molecule so that it can be placed and observed in the vacuum. The contrast generated by the molecule itself is normally inadequate for direct observation in the electron microscope. Various contrasting methods have been developed. Negative staining with heavy metal salts such as uranyl acetate produces a high contrast and prevents the molecule from collapsing. Instead of the molecule, with its interior density variations, only a cast of the exterior surface of the molecule is imaged and only its shape can be reconstructed. To some extent, a stain may penetrate into crevices, but this does not change the fact that only the boundary of the molecule is visualized. As sophisticated averaging methods were developed, it became possible to make sense of the faint images produced by the molecule itself, but biologically relevant information could be obtained only after methods were found to sustain the molecule in a medium that closely approximates the aqueous environment; these methods are embedment in glucose, tannic acid, and vitreous ice. The chapter also describes gold-labeling techniques that are crucial in 3D electron microscopy." @default.
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- W2505111835 date "1996-01-01" @default.
- W2505111835 modified "2023-10-17" @default.
- W2505111835 title "Electron Microscopy of Macromolecular Assemblies" @default.
- W2505111835 doi "https://doi.org/10.1016/b978-012265040-6/50002-3" @default.
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