期刊
ANALYTICAL CHEMISTRY
卷 89, 期 22, 页码 12168-12175出版社
AMER CHEMICAL SOC
DOI: 10.1021/acs.analchem.7b02817
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资金
- U.S. National Institutes of Health Office of Research Infrastructure Programs [P40 OD010440]
- Victorian Government's Operational Infrastructure Support Program
- University of Melbourne Research Grant Support Scheme
- Australian Research Council [DP130100357]
- Multimodal Australian ScienceS Imaging and Visualisation Environment (MASSIVE)
Analytical approaches that preserve the endogenous state of the examined system are essential for the in vivo study of bioinorganics. X-ray fluorescence microscopy of biological samples can map elements in vivo at subcellular resolutions in tissue samples and multicellular organisms. However, X-ray irradiation induces modifications that accumulate with dose. Consequently, the utility of X-ray fluorescence microscopy is intrinsically limited by the radiation damage it causes and the degree to which it alters the target features of interest. Identification of the dose threshold, below which the integrity of the specimen and its elemental distribution is preserved, is required to ensure valid interpretation of concentrations. Here we use the nematode, Caenorhabditis elegans, to explore these issues using three chemical-free specimen preparations: lyophilization, cryofixation, and live. We develop quantitative methods for investigating damage and present dose limits for each preparation pertaining to the micrometer-scale spatial distribution of specific analytes (potassium, calcium, manganese, iron, and zinc), and discuss dose-appropriate guidelines for X-ray fluorescence microscopy of microscale biological samples.
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