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Edited by Margaret M. Murnane, University of Colorado at Boulder, Boulder, CO, and approved March 11, 2010 (received for review September 22, 2009)
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.
Footnotes
- 1To whom correspondence should be addressed. E-mail: Johanna.Nelson{at}stonybrook.edu.
Author contributions: J.K. and C.J. designed research; J.N., X.H., S.M., and J.J.T. performed research; A.M.N. contributed new reagents; J.S. contributed analytic tools; J.N. and D.S. analyzed data; and J.N. and C.J.J. wrote the paper.
The authors declare no conflict of interest.
This article is a PNAS Direct Submission.
This article contains supporting information online at www.pnas.org/cgi/content/full/0910874107/DCSupplemental.
High-resolution x-ray diffraction microscopy of specifically labeled yeast cells
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