Structural analysis of multicellular organisms with cryo-electron tomography.

Harapin, Jan; Börmel, Mandy; Sapra, K Tanuj; Brunner, Damian; Kaech, Andres; Medalia, Ohad · Nat Methods · 2015

basic_science · Level V

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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.

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