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Synchrotron radiation spectroscopic techniques, such as X-ray absorption fine structure (XAFS) has been recognized as a powerful structural characterization method. Its most striking advantages are its element-specificity, sensitivity to the short-range order (typically several Å) and chemical state. Nowadays, XAFS is routinely used in a wide range of scientific fields, including physics, chemistry, biology, materials sciences, environmental sciences, and so on. In this chapter, the synchrotron radiation XAFS technique will be introduced. The advantages of using XAFS for the fine structure characterization of two-dimensional nanomaterials will be discussed. Recent research progress in the field will be reviewed. XAFS provides quantitative structural information at an atomic scale around the specific atom, i.e. near-neighbor species and distance, coordination number, fluctuation in bond distance for the two-dimensional nanomaterials. The distance determination can be accurate to about 0.01 Å or better. An outlook of time- and spatial-resolved synchrotron radiation techniques for two-dimensional nanomaterials will also be provided.

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