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Research Article

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, and Chris Jacobsen
PNAS April 20, 2010 107 (16) 7235-7239; https://doi.org/10.1073/pnas.0910874107
Johanna Nelson
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  • For correspondence: Johanna.Nelson@stonybrook.edu
Xiaojing Huang
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Jan Steinbrener
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David Shapiro
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Janos Kirz
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Stefano Marchesini
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Aaron M. Neiman
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Joshua J. Turner
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Chris Jacobsen
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  1. Edited by Margaret M. Murnane, University of Colorado at Boulder, Boulder, CO, and approved March 11, 2010 (received for review September 22, 2009)

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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.

  • coherent imaging
  • immunogold labeling

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.

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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
Proceedings of the National Academy of Sciences Apr 2010, 107 (16) 7235-7239; DOI: 10.1073/pnas.0910874107

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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
Proceedings of the National Academy of Sciences Apr 2010, 107 (16) 7235-7239; DOI: 10.1073/pnas.0910874107
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Proceedings of the National Academy of Sciences: 107 (16)
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