title: High-resolution x-ray diffraction microscopy of specifically labeled yeast cells.
abstract: X-ray diffraction microscopy complements other x-ray microscopy methods by being free of lens-imposed radiation dose and resolution limits, and it allows for high-resolution imaging of biological specimens too thick to be viewed by electron microscopy. We report here the highest resolution (11-13 nm) x-ray diffraction micrograph of biological specimens, and a demonstration of molecular-specific gold labeling at different depths within cells via through-focus propagation of the reconstructed wavefield. The lectin concanavalin A conjugated to colloidal gold particles was used to label the alpha-mannan sugar in the cell wall of the yeast Saccharomyces cerevisiae. Cells were plunge-frozen in liquid ethane and freeze-dried, after which they were imaged whole using x-ray diffraction microscopy at 750 eV photon energy.
authors.affiliation: Department of Physics and Astronomy, Stony Brook University, Stony Brook, NY 11794-3800, USA. Johanna.Nelson@stonybrook.edu
authors.forename: Johanna
authors.initials: J
authors.lastname: Nelson
authors.forename: Xiaojing
authors.initials: X
authors.lastname: Huang
authors.forename: Jan
authors.initials: J
authors.lastname: Steinbrener
authors.forename: David
authors.initials: D
authors.lastname: Shapiro
authors.forename: Janos
authors.initials: J
authors.lastname: Kirz
authors.forename: Stefano
authors.initials: S
authors.lastname: Marchesini
authors.forename: Aaron M
authors.initials: AM
authors.lastname: Neiman
authors.forename: Joshua J
authors.initials: JJ
authors.lastname: Turner
authors.forename: Chris
authors.initials: C
authors.lastname: Jacobsen
mesh_terms: D003102:Colloids / Q000737:chemistry; D006046:Gold / Q000737:chemistry; D021621:Imaging, Three-Dimensional / Q000379:methods; D008027:Light; D008854:Microscopy, Electron / Q000379:methods; D008855:Microscopy, Electron, Scanning / Q000379:methods; D009154:Mutation; D016011:Normal Distribution; D012097:Repressor Proteins / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics; D012542:Scattering, Radiation; D012984:Software; D014961:X-Ray Diffraction / Q000379:methods*
chemical_list: D003102:Colloids; D012097:Repressor Proteins; D029701:Saccharomyces cerevisiae Proteins; C508130:Whi5 protein, S cerevisiae; D006046:Gold