For analysis of liver histopathology by light microscopy, formalin-fixed paraffin-embedded liver sections were cut at 5 μm and stained with hematoxylin and eosin (H&E), sirius red, or immunohistochemically for cytokeratin-19 (CK-19) antigen by the Michigan State University Investigative Histopathology Laboratory as described previously (18 (link),31 (link)). Hepatocellular necrosis was quantified in a masked fashion by examining 5 representative 40X H&E-stained liver sections from each animal and expressed as percent of total tissue area. For quantification of sirius red (collagen deposits) and CK-19 staining (BDECs), images of stained liver sections were captured using a Virtual Slide System VS110 (Olympus, Hicksville, NY) with a 20X objective. The area of positive sirius red and CK-19 staining in at least 200 images per tissue was determined in an automated and unbiased fashion using a batch macro and the color de-convolution tool in ImageJ. Serum activity of alanine aminotransferase (ALT) was determined using commercial reagents (Infinity ALT/GPT Thermo Fisher, Waltham, MA).
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Comprehensive Liver Histopathology Analysis
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Corresponding Organization : Michigan State University
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Variable analysis
independent variables
- None explicitly mentioned
- Hepatocellular necrosis (percent of total tissue area)
- Sirius red staining (collagen deposits, area of positive staining)
- CK-19 staining (BDEC, area of positive staining)
- Serum activity of alanine aminotransferase (ALT)
- Liver sections were formalin-fixed and paraffin-embedded
- Liver sections were cut at 5 μm thickness
- Liver sections were stained with hematoxylin and eosin (H&E), sirius red, or immunohistochemically for cytokeratin-19 (CK-19) antigen
- No positive or negative controls were explicitly mentioned in the information provided.
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