Structural analysis of multicellular organisms with cryo-electron tomography

Nat Methods. 2015 Jul;12(7):634-6. doi: 10.1038/nmeth.3401. Epub 2015 May 11.

Abstract

We developed a method for visualizing tissues from multicellular organisms using cryo-electron tomography. Our protocol involves vitrifying samples with high-pressure freezing, thinning them with cryo-FIB-SEM (focused-ion-beam scanning electron microscopy) and applying fiducial gold markers under cryogenic conditions to the lamellae post-milling. We applied this protocol to acquire tomograms of vitrified Caenorhabditis elegans embryos and worms, which showed the intracellular organization of selected tissues at particular developmental stages in otherwise intact specimens.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Caenorhabditis elegans / embryology
  • Caenorhabditis elegans / ultrastructure
  • Cryoelectron Microscopy / methods*
  • Electron Microscope Tomography / methods*
  • Microscopy, Electron, Scanning