High-resolution x-ray diffraction microscopy of specifically labeled yeast cells

Johanna Nelson*, Xiaojing Huang, Jan Steinbrener, David Shapiro, Janos Kirz, Stefano Marchesini, Aaron M. Neiman, Joshua J. Turner, Chris Jacobsen

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

86 Scopus citations

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 α-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.

Original languageEnglish (US)
Pages (from-to)7235-7239
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume107
Issue number16
DOIs
StatePublished - Apr 20 2010

Keywords

  • Coherent imaging
  • Immunogold labeling

ASJC Scopus subject areas

  • General

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