Abstract
A workflow was developed and implemented in a software tool for the automated combination of spatially resolved laser ablation-inductively coupled plasma-mass spectrometry (LA-ICP-MS) data and data on the morphology of the biological tissue. Making use of a recently published biological annotation software, FishImager automatically assigns the biological feature as regions of interest (ROIs) and overlays them with the quantitative LA-ICP-MS data. Furthermore, statistical tools including cluster algorithms can be applied to the elemental intensity data and directly compared with the ROIs. This is effectively visualized in heatmaps. This allows gaining statistical significance on distribution and co-localization patterns. Finally, the biological functions of the assigned ROIs can then be easily linked with elemental distributions. We demonstrate the versatility of FishImager with quantitative LA-ICP-MS data of the zebrafish embryo tissue. The distribution of natural elements and xenobiotics is analyzed and discussed. With the help of FishImager, it was possible to identify compartments affected by toxicity effects or biological mechanisms to eliminate the xenobiotic. The presented workflow can be used for clinical and ecotoxicological testing, for example. Ultimately, it is a tool to simplify and reproduce interpretations of imaging LA-ICP-MS data in many applications.Graphical abstract
Highlights
The zebrafish embryo (ZFE, Danio rerio) is an important model organism and used in various research fields such as developmental biology, biomedical research, andtoxicology [1]
regions of interest (ROIs) are often manually assigned in the data processing software to the LA-ICPMS results and compared [20, 21]
Automated identification of ROIs based on sample features can be performed in application-specific software, but these results cannot be combined with the LA-ICP-MS data [17, 22]
Summary
The zebrafish embryo (ZFE, Danio rerio) is an important model organism and used in various research fields such as developmental biology, biomedical research, and (eco)toxicology [1]. One versatile quantitative elemental imaging technique for biological tissues is laser ablation-inductively coupled plasma-mass spectrometry (LAICP-MS). It is applied in various applications, such as determining the distribution of anti-cancer compounds in tumor tissue and nanoparticles in organisms, or the (immune) response on (i) Quality controls like measurement uncertainty and validation parameters such as limit of detection and quantification, linearity of the calibration range, precision, and accuracy. (ii) Tools for statistical analyses such as cluster analysis and principal component analysis These have been recently included in the software HDIP (Hierarchical Data Format version 5-image processing) [19]. The establishment of quantitative LA-ICP-MS image analysis for diagnostics in medicine, pharmacology studies, and (eco)toxicological studies requires an increased effort to generate reproducible data, to link them with data from other analyses, and with morphology and function, for example
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