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Orcein stain for copper-binding protein and immunohistochemistry for cytokeratin 7 (K7) are commonly used to detect signs of chronic cholestasis in the context of chronic biliary disease, particularly at an early stage when bilirubinostasis is lacking and in the non-icteric phase of the disease.1 2 Copper-binding protein deposits and K7 expression by periportal hepatocytes occur in a patchy and heterogenous fashion. In a recent study, we have observed that more than half of 82 periportal regions in 12 liver biopsies from patients with chronic cholangiopathies did not show any stain for copper, copper-binding protein or K7.3 Only a minority (9%) of these periportal regions were positive for rhodanine, orcein and K7 stains. This observation was based on comparing three sections at different levels from each biopsy used for the orcein, rhodanine and K7 stains. Although these sections were cut serially, they were not suitable to investigate whether copper-binding protein deposits and K7 expression were present in the same hepatocytes. We therefore designed a protocol of sequential histochemical (orcein) and immunohistochemical (K7) staining on a single-tissue …
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