Structural studies by electron tomography: from cells to molecules.
Electron tomography (ET) is uniquely suited to obtain three-dimensional reconstructions of pleomorphic structures, such as cells, organelles or supramolecular assemblies. Although the principles of ET have been known for decades, its use has gathered momentum only in recent years, thanks to technological advances and its combination with improved specimen preparation techniques. The rapid freezing/freeze-substitution preparation is applicable to whole cells and tissues, and it is the method of choice for ET investigations of cellular ultrastructure. The frozen-hydrated preparation provides the best possible structural preservation and allows the imaging of molecules, complexes, and supramolecular assemblies in their native state and their natural environment. Devoid of staining and chemical fixation artifacts, cryo-ET provides a faithful representation of both the surface and internal structure of molecules. In combination with advanced computational methods, such as molecular identification based on pattern recognition techniques, cryo-ET is currently the most promising approach to comprehensively map macromolecular architecture inside cellular tomograms.
Lucić
Vladan
V
Department of Structural Biology, Max Planck Institute of Biochemistry, D-82152 Martinsried, Germany. vladan@biochem.mpg.de
Förster
Friedrich
F
Baumeister
Wolfgang
W
D000465:Algorithms; D000818:Animals; D002477:Cells / Q000648:ultrastructure; D020285:Cryoelectron Microscopy / Q000379:methods; D005583:Fourier Analysis; D007091:Image Processing, Computer-Assisted / Q000379:methods / Q000706:statistics & numerical data; D046911:Macromolecular Substances; D015394:Molecular Structure; D015388:Organelles / Q000648:ultrastructure; D014054:Tomography / Q000379:methods* / Q000706:statistics & numerical data
D046911:Macromolecular Substances