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Extending the methodology of X-ray crystallography to allow imaging of micrometre-sized non-crystalline specimens

Abstract

The contrast and penetrating power afforded by soft X-rays when they interact with matter makes this form of radiation ideal for studying micrometre-sized objects1,2. But although soft X-rays areuseful for probing detail too fine for visible light microscopy in specimens too thick for electron microscopy, the highest-resolution applications of X-ray imaging have been traditionally limited to crystalline samples. Here we demonstrate imaging (at 75 nm resolution) of a non-crystalline sample, consisting of an array of gold dots, by measuring the soft X-ray diffraction pattern from which an image can be reconstructed. The crystallographic phase problem3 — the usually unavoidable loss of phase information in the diffraction intensity — is overcome by oversampling4 the diffraction pattern, and the image is obtained using an iterative algorithm5. Our X-ray microscopy technique requires no high-resolution X-ray optical elements or detectors. We believe that resolutions of 10–20 nm should be achievable; this would provide an imaging resolution about 100 times lower than that attainable with conventional X-ray crystallography, but our method is applicable to structures roughly 100 times larger. This latter feature may facilitate the imaging of small whole cells or large subcellular structures in cell biology.

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Figure 1: A scanning electron microscope image of the specimen.
Figure 2: A diffraction pattern of the specimen (using a logarithmic intensity scale).
Figure 3
Figure 4

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Acknowledgements

The decision to try oversampling as a phasing technique was arrived at in a conversation in the late 1980s with G. Bricogne. W. Yun and H. N. Chapman also participated in early parts of this experiment. We thank C. Jacobsen for help and advice, especially with the numerical reconstruction, and we thank him and M. Howells for use of the apparatus20 in which the exposures were made; we also thank S. Wirick for help with data acquisition. P.C. thanks the Leverhulme Trust Great Britain for supporting the nanofabrication programme at King's College, London. This work was performed at the National Synchrotron Light Source, which is supported by the US Department of Energy. Our work was supported in part by the US Department of Energy.

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Correspondence to Jianwei Miao.

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Miao, J., Charalambous, P., Kirz, J. et al. Extending the methodology of X-ray crystallography to allow imaging of micrometre-sized non-crystalline specimens. Nature 400, 342–344 (1999). https://doi.org/10.1038/22498

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