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  Integrative Approaches for Cellular Cryo-electron Tomography: Correlative Imaging and Focused Ion Beam Micromachining

Rigort, A., Villa, E., Bäuerlein, F. J. B., Engel, B. D., & Plitzko, J. M. (2012). Integrative Approaches for Cellular Cryo-electron Tomography: Correlative Imaging and Focused Ion Beam Micromachining. In Methods in Cell Biology; Vol. 111 (pp. 259-281). San Diego: Academic Press.

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 Urheber:
Rigort, Alexander1, Autor           
Villa, Elizabeth1, Autor           
Bäuerlein, Felix Jan Benedict1, Autor           
Engel, Benjamin D.1, Autor           
Plitzko, Jürgen M.1, Autor           
Affiliations:
1Baumeister, Wolfgang / Molecular Structural Biology, Max Planck Institute of Biochemistry, Max Planck Society, ou_1565142              

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 Zusammenfassung: The application of cryo-electron tomography to cells and tissues is commonly referred to as “cellular tomography,” and enables visualization of the supramolecular architecture of cells in a near-native state. However, in order to access structural features hidden deep inside cellular volumes, it is necessary to use hybrid techniques to identify and localize features of interest and prepare such regions for subsequent analysis by transmission electron microscopy. We present a workflow that integrates different approaches: (1) correlative cryo-fluorescence microscopy to localize features within frozen-hydrated cells, (2) focused ion beam milling to thin these specimens in a targeted manner, and (3) cryo-electron tomography to provide detailed information about the cellular ultrastructure of thinned samples. We describe the combined use of these techniques and the instrumentation required to enable cryo-electron tomography for a vast range of cellular samples.

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Sprache(n): eng - English
 Datum: 2012
 Publikationsstatus: Erschienen
 Seiten: -
 Ort, Verlag, Ausgabe: -
 Inhaltsverzeichnis: -
 Art der Begutachtung: Expertenbegutachtung
 Identifikatoren: DOI: 10.1016/B978-0-12-416026-2.00014-5
 Art des Abschluß: -

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Titel: Methods in Cell Biology; Vol. 111
  Alternativer Titel : Chapter 14
Genre der Quelle: Reihe
 Urheber:
Affiliations:
Ort, Verlag, Ausgabe: San Diego : Academic Press
Seiten: - Band / Heft: 111 Artikelnummer: - Start- / Endseite: 259 - 281 Identifikator: ISSN: 0091-679X