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Nat Methods. 2014 Jul;11(7):737-9. doi: 10.1038/nmeth.2961. Epub 2014 May 11.

Correlated cryogenic photoactivated localization microscopy and cryo-electron tomography.

Author information

1
1] Division of Biology and Biological Engineering, California Institute of Technology, Pasadena, California, USA. [2] Howard Hughes Medical Institute, Pasadena, California, USA.
2
Division of Biology and Biological Engineering, California Institute of Technology, Pasadena, California, USA.
3
Max Planck Institute for Terrestrial Microbiology, Marburg, Germany.

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.

PMID:
24813625
PMCID:
PMC4081473
DOI:
10.1038/nmeth.2961
[Indexed for MEDLINE]
Free PMC Article

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