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Updated: Sep 8, 2025

Biological Samples Preparation for Speciation at Cryogenic Temperature using High-Resolution X-Ray Absorption Spectroscopy
Published on: May 27, 2022
Biological Samples Preparation for Speciation at Cryogenic Temperature using High-Resolution X-Ray Absorption
Caroline Bissardon1, Marie-Pierre Isaure2, Emmanuel Lesuisse3
1University Grenoble Alpes, INSERM UA7, Synchrotron Radiation for Biomedicine (STROBE).
Abstract:
The study of elements with X-ray absorption spectroscopy (XAS) is of particular interest when studying the role of metals in biological systems. Sample preparation is a key and often complex procedure, particularly for biological samples. Although X-ray speciation techniques are widely used, no detailed protocol has been yet disseminated for users of the technique. Further, chemical state modification is of concern, and cryo-based techniques are recommended to analyze the biological samples in their near-native hydrated state to provide the maximum preservation of chemical integrity of the cells or tissues. Here, we propose a cellular preparation protocol based on cryo-preserved samples. It is demonstrated in a high energy resolution fluorescence detected X-ray absorption spectroscopy study of selenium in cancer cells and a study of iron in phytoplankton. This protocol can be used with other biological samples and other X-ray techniques that can be damaged by irradiation.
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