Characterization of Liver Malignant Lesions

Five-µm liver sections were colored with the Masson’s Trichrome (MT) stain. Tissue sections were also processed for immunohistological analysis with antibodies against Melan-A (clone A103, Agilent Dako, Mississauga, ON, Canada) and αSMA (clone 1A4, Agilent Dako) in a Dako Autostainer Plus Link, according to the manufacturer’s protocol using the EnVision peroxidase procedure with the DAB or Magenta chromogen (Agilent Dako) [45 (link)]. The demasking was done at high pH. Secondary antibody without primary antibody was used as negative control, and positive controls were performed with skin and muscle tissues. Sections were digitized with a slide scanner (NanoZoomer 2.0HT; Hamamatsu Photonics, Bridgewater, NJ, USA), and images were analyzed with the NDP.view 2 software (Hamamatsu Photonics). Liver malignant lesions were counted on MT-stained slides using the measure feature of NDP.view 2. They were then categorized according to their size (<50 µm, 50–500 µm, >500 µm in diameter) [23 (link)].