Ventana medical system benchmark xt ish staining module

Four-µm thick tissue sections of INCPH and control cases were incubated with the primary antibodies CD34 (qbend/10, Novocastra, 1 h incubation) and Glutamine synthetase (6/GS, Biocare, 1 h incubation), by using an automated staining module (Ventana Medical System-Benchmark XT/ISH Staining Module, Roche, Switzerland). Results were evaluated by two experienced liver pathologists (MG, MB).

Paraffin blocks that were representative of all histologic patterns present together were selected for immunohistochemical analysis. For this purpose, one to two blocks per case were used. Immunostaining for MUC1, MUC2, MUC5AC, and MUC6 proteins was performed on 4-µm tissue sections, using an automated staining module (Ventana Medical System-Benchmark XT/ISH Staining Module, Roche, Switzerland). Tissue sections were incubated with the primary antibodies with different incubation periods and different dilutions. Anti-MUC1 antibody (Ma695, 1:100, Leica/Novocastra), anti-MUC2 antibody (Ccp58, 1:50, Leica/Novocastra), anti-MUC5AC antibody (CLH2, 1:50, Leica/Novocastra) and anti-MUC6 antibody (CLH5, 1:50, Leica/Novocastra) were used.