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Cryo-electron microscopy of vitreous sections
Authors:Al-Amoudi Ashraf  Chang Jiin-Ju  Leforestier Amélie  McDowall Alasdair  Salamin Laurée Michel  Norlén Lars P O  Richter Karsten  Blanc Nathalie Sartori  Studer Daniel  Dubochet Jacques
Institution:Laboratoire d'Analyse Ultrastructurale, Batiment de Biologie, Université de Lausanne, Lausanne, Switzerland.
Abstract:Since the beginning of the 1980s, cryo-electron microscopy of a thin film of vitrified aqueous suspension has made it possible to observe biological particles in their native state, in the absence of the usual artefacts of dehydration and staining. Combined with 3-d reconstruction, it has become an important tool for structural molecular biology. Larger objects such as cells and tissues cannot generally be squeezed in a thin enough film. Cryo-electron microscopy of vitreous sections (CEMOVIS) provides then a solution. It requires vitrification of a sizable piece of biological material and cutting it into ultrathin sections, which are observed in the vitrified state. Each of these operations raises serious difficulties that have now been overcome. In general, the native state seen with CEMOVIS is very different from what has been seen before and it is seen in more detail. CEMOVIS will give its full potential when combined with computerized electron tomography for 3-d reconstruction.
Keywords:bacterial envelope  desmosome  high-pressure freezing  intermediate filament  vitrification
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