Ventana discovery ultra ihc auto stainer

The dual IHC (ER/Ki67 and Ki67/PR) was performed by the Pathology Solid Tumor Core at City of Hope using Ventana Discovery Ultra IHC Auto Stainer (Roche Diagnostics). Primary antibodies used for the immunostaining include human ERα rabbit monoclonal antibody (Roche Diagnostics, 790−4325), human PR rabbit monoclonal antibody (Roche Diagnostics, 790−4296) and human Ki67 rabbit monoclonal antibody (Roche Diagnostics, 790−4286). Images were captured with VENTANA iScan HT (Roche Diagnostics) and analysed using the Cell Detection and Cell Classification functions implemented in Qupath software.
²⁴ (link) Staining protocol details are described in Supplementary Methods.

The fourth mouse mammary gland of eight-weeks-old C57BL/6J mice were collected with skin and fixed with 10% buffered formalin. After embedding in paraffin, the cross-sections of the tissue were prepared and used for H&E staining and immunohistochemistry. Immunohistochemistry was performed by the Pathology Solid Tumor Core at City of Hope using Ventana Discovery Ultra IHC Auto Stainer (Roche Diagnostics, Indianapolis, IN, United States). Heat-mediated antigen retrieval was performed using Cell Conditioning Buffer 1 (Roche Diagnostics; pH 8.5) for an hour. Antibodies used for the immunostaining included: ERα rabbit polyclonal antibody (06–935, Millipore Sigma; 1:400), anti-PDGFRα rabbit monoclonal antibody (ab134123, Abcam, Cambridge, United Kingdom; 1:50), and anti-DPP4 rabbit monoclonal antibody (ab187048, Abcam; 1:500). Images were captured on the Zeiss Observer II (Carl Zeiss, Oberkochen, Germany; for H&E staining), VENTANA iScan HT (Roche Diagnostics; for PDGFRα and DPP4 immunohistochemistry) or Nano Zoomer S360 (HAMAMATSU PHOTONICS, Shizuoka, Japan; for ERα immunohistochemistry).