Electron Tomography of Frozen-hydrated Sections of Cells and Tissues

摘要: The technique of cryoelectron tomography frozen-hydrated biological specimens is opening a new window on cellular structure and organization. This imaging method provides full 3D structural information at much higher resolution (typically 5–10 nm) than attainable by light microscopy, can be applied to cells organelles that are maintained in state as close native achieved currently electron microscopy. Not only used visualize directly extended structures, such membranes cytoskeleton, but it also provide maps the location, orientation and, perhaps, conformation large macromolecular complexes, cell’s ‘molecular machinery’. complements coming from single-particle microscopy (Frank et al., 1996, 2006) X-ray crystallography, about subnanometer same molecular assemblies after isolation. As with studies using smaller 1 µ size prepared for plunge-freezing (Dubochet 1988). Cells or rapidly frozen an microscope grid thin layers glass-like, amorphous ice, without formation ice crystals would otherwise disrupt fine (Kellenberger, 1987). Specimens imaged directly, chemical fixation, dehydration staining heavy metals.