X-ray fluorescence microscopy methods for biological tissues

Synchrotron-based X-ray fluorescence microscopy is a flexible tool for identifying the distribution of trace elements in biological specimens across a broad range of sample sizes. The technique is not particularly limited by sample type and can be performed on ancient fossils, fixed or fresh tissue specimens, and in some cases even live tissue and live cells can be studied. The technique can also be expanded to provide chemical specificity to elemental maps, either at individual points of interest in a map or across a large field of view. While virtually any sample type can be characterized with X-ray fluorescence microscopy, common biological sample preparation methods (often borrowed from other fields, such as histology) can lead to unforeseen pitfalls, resulting in altered element distributions and concentrations. A general overview of sample preparation and data-acquisition methods for X-ray fluorescence microscopy is presented, along with outlining the general approach for applying this technique to a new field of investigation for prospective new users. Considerations for improving data acquisition and quality are reviewed as well as the effects of sample preparation, with a particular focus on soft tissues. The effects of common sample pretreatment steps as well as the underlying factors that govern which, and to what extent, specific elements are likely to be altered are reviewed along with common artifacts observed in X-ray fluorescence microscopy data.

Automated summary

Synchrotron-based X-ray fluorescence microscopy is a versatile technique for identifying trace element distribution in biological samples of various sizes. It can be used on different types of samples, including ancient fossils, fixed or fresh tissue specimens, and even live tissue and live cells. The technique can also provide chemical specificity to elemental maps, either at individual points of interest or across a large field of view. However, common sample preparation methods in biology can lead to unexpected problems, such as altered element distributions and concentrations.