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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.

SUBMITTER: Nelson J 

PROVIDER: S-EPMC2867732 | biostudies-literature | 2010 Apr

REPOSITORIES: biostudies-literature

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

Nelson Johanna J   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  

Proceedings of the National Academy of Sciences of the United States of America 20100405 16


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 wave  ...[more]

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