High-resolution x-ray diffraction microscopy of specifically labeled yeast cells.
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.
Nelson
Johanna
J
Department of Physics and Astronomy, Stony Brook University, Stony Brook, NY 11794-3800, USA. Johanna.Nelson@stonybrook.edu
Huang
Xiaojing
X
Steinbrener
Jan
J
Shapiro
David
D
Kirz
Janos
J
Marchesini
Stefano
S
Neiman
Aaron M
AM
Turner
Joshua J
JJ
Jacobsen
Chris
C
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*
D003102:Colloids; D012097:Repressor Proteins; D029701:Saccharomyces cerevisiae Proteins; C508130:Whi5 protein, S cerevisiae; D006046:Gold