Electron microscopic detection of copper in the liver of two patients withmorbus Wilson by EELS and EDX

A 20-year-old male patient with morbus Wilson was liver transplanted becaus e of terminal failure of liver function. The explanted liver showed a stron g macronodular cirrhosis as typically seen in Wilson disease. There were vi sible granular accumulations in the hepatocytes after the rubeanic acid or rhodanine method for histochemical detection of copper. The electron micros copic studies on ultrathin sections revealed numerous electron-dense lysoso mes and residual bodies. The elemental analysis in transmission electron mi croscope (TEM) with electron energy loss spectroscopy (EELS) and in scannin g electron microscope (SEM) with energy dispersive x-ray analysis (EDX) sho wed copper-specific signals of electron-dense accumulations inside these da rk lysosomes and residual bodies. In a second case, Wilson disease was diag nosed after autopsy of a 31-year-old patient by liver electron microscopy a nd EELS; strong electron-dense lysosomes and residual bodies with positive copper signals were found inside hepatocytes. for negative control, hepatoc ytes with iron accumulation after idiopathic hemochromatosis and liver cirr hosis were also analyzed by EELS in TEM, which showed strong iron, but only a few or no copper signals. Atomic absorption spectroscopy (AAS) in 16 liv er samples of healthy and cirrhotic liver revealed only in both cases of Wi lson disease a strong increased copper concentration higher than 100 mug Cu /g. The electron microscopic detection of copper-containing hepatocytic lys osomes is helpful for rile diagnosis of early stages of Wilson disease in a ddition to the quantification of hepatic copper by AAS.