title: Structural analysis of multicellular organisms with cryo-electron tomography.
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.
authors.affiliation: Department of Biochemistry, University of Zurich, Zurich, Switzerland.
authors.forename: Jan
authors.initials: J
authors.lastname: Harapin
authors.affiliation: 1] Department of Biochemistry, University of Zurich, Zurich, Switzerland. [2] Institute of Molecular Life Sciences, University of Zurich, Zurich, Switzerland.
authors.forename: Mandy
authors.initials: M
authors.lastname: Börmel
authors.affiliation: Department of Biochemistry, University of Zurich, Zurich, Switzerland.
authors.forename: K Tanuj
authors.initials: KT
authors.lastname: Sapra
authors.affiliation: Institute of Molecular Life Sciences, University of Zurich, Zurich, Switzerland.
authors.forename: Damian
authors.initials: D
authors.lastname: Brunner
authors.affiliation: Center for Microscopy and Image Analysis, University of Zurich, Zurich, Switzerland.
authors.forename: Andres
authors.initials: A
authors.lastname: Kaech
authors.affiliation: 1] Department of Biochemistry, University of Zurich, Zurich, Switzerland. [2] Department of Life Sciences in the Negev, Ben-Gurion University, Beer-Sheva, Israel. [3] National Institute for Biotechnology in the Negev, Ben-Gurion University, Beer-Sheva, Israel.
authors.forename: Ohad
authors.initials: O
authors.lastname: Medalia
mesh_terms: D000818:Animals; D017173:Caenorhabditis elegans / Q000196:embryology / Q000648:ultrastructure; D020285:Cryoelectron Microscopy / Q000379:methods*; D055032:Electron Microscope Tomography / Q000379:methods*; D008855:Microscopy, Electron, Scanning