Iron, copper, zinc and bromine mapping in cirrhotic liver slices from patients with hemochromatosis studied by microscopic synchrotron radiation X-ray fluorescence analysis in continuous scanning mode

Abstract

Iron (Fe) and copper (Cu) are essential metals in physiological cell metabolism. While Fe is easy to determine biochemically in histological slices, Cu and zinc (Zn) distribution is frequently critical in confirming the presence of an overload in disturbed Fe/Cu metabolism. To analyze Fe, Cu and Zn in a near histological resolution, energy dispersive microscopic synchrotron radiation X-ray fluorescence was applied.In normal liver tissue, after fixation and imbedding in paraffin, mean Fe, Cu and Zn concentrations were 152 ± 54, 20.1 ± 4.3 and 88.919.5 μg/g sample weight, respectively. No substantial, characteristic differences in their distribution were found in the two-dimensional scans. In slices from patients with hemochromatosis mean Fe, Cu and Zn concentrations were 1102 ± 539, 35.9 ± 14.6 and 27.2 ± 6.7 μg/g sample weight, respectively. Additionally, a significant decrease in phosphorus and sulphur concentrations existed. An increased Cu around cirrhotic regenerations nodules is mostly associated with a lymphocytic infiltration in this region. Analyzing concentrations of Fe in different regions of the samples show a clear negative dependency between Fe and Cu, Cu and Zn, but a positive one between Fe and Zn.Conclusion: With a focal beam size of 15 μm in diameter a resolution of the elemental distribution was achieved which is widely comparable with stained histological slices (20× light microscope). The analysis of simultaneous determined elements reveals metabolic differences between Fe, Cu and Zn in liver tissue from patients with hemochromatosis.

Quantitative trace element mapping in liver tissue from patients with Wilson`s disease determined by micro X-ray fluorescence

“…The technical setup used for the performed measurements is extensively described in recent papers (18,24). In short: microscopic synchrotron radiation X-ray fluorescence analysis (µ-SRXRF) measurements were performed at beamline L at the DORIS III storage ring at HASYLAB/DESY in Hamburg.…”

“…Among histochemical stains for Cu Timm's silver stain seems to be the most sensitive method for the demonstration of copper while Rhodanine and Orcein have been proven to be less sensible in staining either free copper or copper-binding protein corresponding to metallothioneins, respectively (6)(7)(8). For quantitative imaging of metals in tissue electron microprobe (10), laser ablation coupled plasma mass spectroscopy -which is tissue destructive - (11)(12) and micro synchrotron-based X-ray fluorescence (micro-SRXRF) (15)(16)(17)(18)(19) have been introduced.…”

“…Metal and trace element investigations by micro Synchrotron X-ray fluorescence (µ-SRXRF) have increasingly expanded basic knowledge and clinical diagnosis (19) by determining element distribution in a variety of animal and human tissues (14)(15)(16)(17)(18).…”

In Situ Imaging of Metals in Cells and Tissues

“…The interconnection between Fe, Cu, and possibly Zn metabolism407 was the impetus for investigating the interplay between these metals in cirrhotic liver slices from hemochromatosis patients 408. SXRF scans with a focal beam diameter of 15 µm showed a substantial increase in Cu content around cirrhotic regeneration nodules primarily associated with lymphocytic infiltration.…”

Rapid and reliable diagnosis of Wilson disease using X‐ray fluorescence

“…This technique has the advantages of being non‐destructive and highly sensitive even at trace levels without any histochemical staining or elaborate sample preparation, which make it compatible with various medical research areas . XRF analysis has allowed investigation of hepatic iron (Fe) distribution , the distribution of metalloproteins in hepatocellular carcinoma and surrounding tissues as well as elemental mapping (calcium (Ca), iron (Fe), copper (Cu), zinc (Zn) and bromine (Br)) on liver tissue sections from patients with haemochromatosis or cirrhosis . Interestingly, higher accumulation of copper in a mouse animal model of WD has been observed using synchrotron X‐ray absorption and fluorescence microspectroscopy .…”