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Nano-fEM: Protein Localization Using Photo-activated Localization Microscopy and Electron Microscopy
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Biologie
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Journal JoVE Biologie
Nano-fEM: Protein Localization Using Photo-activated Localization Microscopy and Electron Microscopy

Nano-fEM: Protein Localization Using Photo-activated Localization Microscopy and Electron Microscopy

DOI:
10.3791/3995-v

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13:13 min

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December 03, 2012

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, , , , ,
1Department of Biology, Howard Hughes Medical Institute,University of Utah

Chapitres

  • 00:05Titre
  • 01:24High Pressure Freezing
  • 02:45Freeze-substitution
  • 04:27Infiltration and Polymerization
  • 06:15PALM Imaging
  • 08:37SEM Imaging
  • 09:29Aligning PALM and EM Images
  • 11:51Representative Nano-fEM Images
  • 12:39Conclusion

Summary

Traduction automatique

Traduction automatique

We describe a method to localize fluorescently tagged proteins in electron micrographs. Fluorescence is first localized using photo-activated localization microscopy on ultrathin sections. These images are then aligned to electron micrographs of the same section.

Tags

Nano-fEMProtein LocalizationPhoto-activated Localization MicroscopyElectron MicroscopyFluorescence MicroscopyImmuno-electron MicroscopyCorrelative ApproachesFiducial MarkersPlastic EmbeddingDiffraction Limit Of LightNano-resolution Fluorescence Electron Microscopy

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