Cryo transmission electron microscopy (cryoTEM) is used with vitreous ice-embedded samples as a quantitative tool to investigate the assembly, regulation and function of biologically important macromolecular complexes. Despite continuous progress in x-ray crystallographic and NMR methods, it is still difficult using these techniques alone to obtain atomic level structural information about the many complex macromolecular assemblies that govern fundamental cell biological processes. It is now clear that a powerful approach is to combine structural information obtained at different levels. The structural information obtained from cryoTEM and computer based 3-D reconstruction spans the gap between our ability to obtain structural information about large macromolecular assemblies using conventional light and electron microscopy, and high-resolution data obtained from NMR or X-ray crystallography. These data can be combined to build up a detailed picture of the overall architecture of the complexes and the interactions between the components.
The development of an electron cryomicroscopy laboratory is a major component of our ongoing low temperature electron microscopy initiative. The focus of the electron cryomicroscopy laboratory will be analysis of macromolecules frozen in vitreous ice. Another application of the newly installed 200KV electron cryomicroscope will be electron tomography. We will be able to construct three-dimensional models of ultrastructural morphology.
The establishment of the electron cryomicroscopy laboratory is another step in our continuing effort to obtain better morphological preservation with less chemical processing.
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Electron Microscopy Preperation