martes, 23 de agosto de 2011

Cryo-electron tomography

3-D Ultrastructure of O. tauri: Electron Cryotomography of an Entire Eukaryotic Cell

Henderson, Gregory P. and Gan, Lu and Jensen, Grant J. (2007) 3-D Ultrastructure of O. tauri: Electron Cryotomography of an Entire Eukaryotic Cell. PLoS ONE, 2 (8). e749. ISSN 1932-6203 http://resolver.caltech.edu/CaltechAUTHORS:HENplosone07

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Abstract

The hallmark of eukaryotic cells is their segregation of key biological functions into discrete, membrane-bound organelles. Creating accurate models of their ultrastructural complexity has been difficult in part because of the limited resolution of light microscopy and the artifact-prone nature of conventional electron microscopy. Here we explored the potential of the emerging technology electron cryotomography to produce three-dimensional images of an entire eukaryotic cell in a near-native state. Ostreococcus tauri was chosen as the specimen because as a unicellular picoplankton with just one copy of each organelle, it is the smallest known eukaryote and was therefore likely to yield the highest resolution images. Whole cells were imaged at various stages of the cell cycle, yielding 3-D reconstructions of complete chloroplasts, mitochondria, endoplasmic reticula, Golgi bodies, peroxisomes, microtubules, and putative ribosome distributions in-situ. Surprisingly, the nucleus was seen to open long before mitosis, and while one microtubule (or two in some predivisional cells) was consistently present, no mitotic spindle was ever observed, prompting speculation that a single microtubule might be sufficient to segregate multiple chromosomes.

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