Isotope dilution LA-ICP-MS for quantitative imaging of trace elements in mouse brain sections

Isotope dilution (ID) analysis is considered one of the most accurate quantitative methods. However, it has not been widely applied to the quantitative imaging of trace elements in biological samples using laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS), mainly because of difficulties in homogeneously mixing enriched isotopes (the spike) with the sample (e.g., a tissue section). In this study, we present a novel method for the quantitative imaging of trace elements (copper and zinc) in mouse brain sections using ID-LA-ICP-MS. We used an electrospray-based coating device (ECD) to evenly distribute a known amount of the spike (⁶⁵Cu and ⁶⁷Zn) on the sections. The optimal conditions for this process involved evenly distributing the enriched isotopes on mouse brain sections mounted on indium tin oxide (ITO) glass slides using the ECD with the 10 mg g^-1 ɑ-cyano-4-hydroxycinnamic acid (CHCA) in methanol at 80 °C. The mass of the spiked isotopes and the tissue sections on the ITO slides was calculated by weighing them on an analytical balance. Quantitative images of Cu and Zn in Alzheimer's disease (AD) mouse brain sections were obtained using ID-LA-ICP-MS. These imaging results showed that Cu and Zn concentrations in various brain regions typically ranged from 10 to 25 μg g^-1 and 30-80 μg g^-1, respectively. But it is worth noting that the hippocampus contained up to 50 μg g^-1 of Zn, while the cerebral cortex and hippocampus had Cu contents as high as 150 μg g^-1. These results were validated by acid digestion and solution analysis with ICP-MS. The novel ID-LA-ICP-MS method provides an accurate and reliable means for quantitative imaging of biological tissue sections.