Introduction to Electron Microscopy
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Transmission electron microscopy (TEM) offers the possibility of visualizing biological structures at resolution well beyond that of light microscopy. Whether you are interested in the ultrastructure of cells and organelles, or in the detailed molecular structure of biological macromolecules, different modalities of TEM can generally be applied to your system of interest.
The lecture reviews the physical principles underlying image formation by the interaction of electrons with matter, introduces you to basic and advanced instruments and to sample preparation techniques. Using a number of biological examples from work in the Nogales lab, the lecture then describes the capabilities of the TEM methodology. Special emphasis is placed on the image processing methods used to obtain three-dimensional information from TEM data.
Eva Nogales is a Howard Hughes Medical Institute investigator, a Professor of Molecular Cell Biology at the University of California, Berkeley, and Senior Faculty Scientist at the Lawrence Berkeley National Laboratory.
She obtained her B.S. degree in physics from the Universidad Autonoma de Madrid (Spain) and did her thesis work at the Synchrotron Radiation Source (U.K.), under the supervision of Joan Bordas, earning a Ph.D. degree from the University of Keele. Her postdoctoral work in Kenneth Downing's group at the Lawrence Berkeley National Laboratory involved the use of electron crystallography to determine the high-resolution structure of tubulin.
The Nogales lab is interested in the structural characterization of complex biological assemblies, their architecture, interactions with different ligands, and the regulation of their function. They are involved in deciphering the molecular bases of cytoskeletal function during cell division and of essential nucleic acid transactions. Towards these aims they use electron microscopy, image analysis, and functional biochemical and biophysical assays.
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Nogales, E. & Grigorieff, N. (2001) "Molecular machines: putting the pieces together", J. Cell Biol. 152, F1-F10.
Leszchiner, A. & Nogales, E. (2007). Visualizing flexibility at molecular resolution: analysis of heterogeneity in single-particle electron microscopy reconstructions. Ann. Rev. Biophys. Biomol. Struct. 36, 43-62.
Electron Crystallography of Biological Macromolecules by Robert
Glaeser, Kenneth Downing, David DeRosier, Wah Chiu and Joachim Frank. Oxford University Press, 2007.
Transmission Electron Microscopy: Physics of Image Formation (Springer
Series in Optical Sciences, Vol 36) by L. Reimer and H. Kohl, 2008
Three-Dimensional Electron Microscopy of Macromolecular Assemblies:
Visualization of Biological Molecules in Their Native State by Joachim Frank. Oxford University Press, 2006.