title: Building Cell Structures in Three Dimensions: Electron Tomography Methods for Budding Yeast.
abstract: Saccharomyces cerevisiaehas been an important model system for numerous cellular, genetic, and molecular studies. However, this small eukaryote presents a challenge for imaging at the electron microscope level. Preparation of yeast using high-pressure freezing followed by freeze-substitution (HPF/FS) results in excellent preservation of cell structure in these difficult-to-fix samples. In particular, cells prepared by HPF/FS can be used for 3D electron tomography (ET) studies where optimum cell preservation is critical. Here, we discuss the advantages of using HPF/FS for ET and show examples of the utility of this method for building yeast cell structures in three dimensions.
authors.affiliation: Boulder Laboratory for 3-D Electron Microscopy of Cells, Department of Molecular, Cellular, and Developmental Biology, University of Colorado, Boulder, Colorado 80309-0347 eileen.otoole@colorado.edu mark.winey@colorado.edu.
authors.forename: Eileen T
authors.initials: ET
authors.lastname: O'Toole
authors.affiliation: Boulder Laboratory for 3-D Electron Microscopy of Cells, Department of Molecular, Cellular, and Developmental Biology, University of Colorado, Boulder, Colorado 80309-0347.
authors.forename: Thomas H
authors.initials: TH
authors.lastname: Giddings
authors.affiliation: Boulder Laboratory for 3-D Electron Microscopy of Cells, Department of Molecular, Cellular, and Developmental Biology, University of Colorado, Boulder, Colorado 80309-0347 eileen.otoole@colorado.edu mark.winey@colorado.edu.
authors.forename: Mark
authors.initials: M
authors.lastname: Winey
mesh_terms: D055032:Electron Microscope Tomography / Q000379:methods*; D017110:Freeze Substitution; D005615:Freezing; D056229:Fungal Structures / Q000648:ultrastructure*; D021621:Imaging, Three-Dimensional / Q000379:methods*; D011309:Preservation, Biological; D012441:Saccharomyces cerevisiae / Q000648:ultrastructure*