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Correlated cryogenic photoactivated localization microscopy and cryo-electron tomography.

ABSTRACT: Cryo-electron tomography (CET) produces three-dimensional images of cells in a near-native state at macromolecular resolution, but identifying structures of interest can be challenging. Here we describe a correlated cryo-PALM (photoactivated localization microscopy)-CET method for localizing objects within cryo-tomograms to beyond the diffraction limit of the light microscope. Using cryo-PALM-CET, we identified multiple and new conformations of the dynamic type VI secretion system in the crowded interior of Myxococcus xanthus.

SUBMITTER: Chang YW 

PROVIDER: S-EPMC4081473 | biostudies-literature | 2014 Jul

REPOSITORIES: biostudies-literature

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Correlated cryogenic photoactivated localization microscopy and cryo-electron tomography.

Chang Yi-Wei YW   Chen Songye S   Tocheva Elitza I EI   Treuner-Lange Anke A   Löbach Stephanie S   Søgaard-Andersen Lotte L   Jensen Grant J GJ  

Nature methods 20140511 7

Cryo-electron tomography (CET) produces three-dimensional images of cells in a near-native state at macromolecular resolution, but identifying structures of interest can be challenging. Here we describe a correlated cryo-PALM (photoactivated localization microscopy)-CET method for localizing objects within cryo-tomograms to beyond the diffraction limit of the light microscope. Using cryo-PALM-CET, we identified multiple and new conformations of the dynamic type VI secretion system in the crowded  ...[more]

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