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Structural analysis of multicellular organisms with cryo-electron tomography


Harapin, Jan; Börmel, Mandy; Sapra, K Tanuj; Brunner, Damian; Kaech, Andres; Medalia, Ohad (2015). Structural analysis of multicellular organisms with cryo-electron tomography. Nature Methods, 12(7):634-636.

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.

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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Additional indexing

Item Type:Journal Article, refereed, original work
Communities & Collections:04 Faculty of Medicine > Department of Biochemistry
07 Faculty of Science > Department of Biochemistry

04 Faculty of Medicine > Center for Microscopy and Image Analysis
Dewey Decimal Classification:570 Life sciences; biology
610 Medicine & health
Language:English
Date:July 2015
Deposited On:24 Sep 2015 10:27
Last Modified:08 Dec 2017 14:06
Publisher:Nature Publishing Group
ISSN:1548-7091
Publisher DOI:https://doi.org/10.1038/nmeth.3401
PubMed ID:25961413

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