Laser ablation-inductively coupled plasma-mass spectrometry for quantitative mapping of the copper distribution in liver tissue sections from mice with liver disease induced by common bile duct ligation

Laser ablation-inductively coupled plasma-mass spectrometry (LA-ICP-MS) was deployed for quantitative mapping of the Cu distribution in cryo-sections of liver tissue from mice with cholestatic liver disease induced via common bile duct ligation (CBDL). Cu distribution maps of the liver sections were obtained from the CBDL-operated mice sacrificed at different time points (2, 4 and 6 weeks) after the surgical intervention and compared with those of the corresponding control (sham-operated) mice. Cu quantification was accomplished versus matrix-matched thin sections of spiked liver tissue homogenates and versus spiked gelatin droplet standards. No statistical differences were obtained between the results using the two calibration approaches and thus, both were considered suitable for quantitative Cu bioimaging of liver cryo-sections. On the basis of practical considerations, i.e. simplicity, low cost and availability of material, spiked gelatin droplet standards are the preferred choice for quantitative determination of the Cu distribution in liver tissue cryo-sections. An inhomogeneous hepatic Cu distribution was observed in the CBDL mice, in contrast to the homogeneous hepatic Cu distribution established for the sham-operated mice. The Cu levels increased with the progression of the disease and a strong accumulation was observed in some necrotic areas. High-resolution LA-ICP-MS bioimaging, using a circular spot size of 2 μm, was suitable for the visualization of the Cu distribution in liver tissue on a (sub-)cellular level. In addition to the quantitative Cu mapping, also the spatial distribution of Zn was monitored in the liver cryo-sections of the control and the 2, 4 and 6 weeks CBDL mice, but in all cases, Zn was homogeneously distributed across the tissue.

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