Slidepath digital imaging hub dih

Immunohistochemical staining was performed using freshly cut 5 micron sections from each site shipped to Stanford University and a commercial antibody for MUC1 (1:50 dilution; SC-7313, Santa Cruz Biotechnology) [20 (link)]. The digital image documentation of all stained slides was performed using the Leica SCN400 scanning system with the SL801 autoloader (Leica Microsystems; Concord, Ontario, Canada) at magnification equivalent to 40x. The images were transferred into the SlidePath digital imaging hub (DIH; Leica Microsystems). In parallel, separate TMA sections were stained with hematoxylin and eosin (H & E) and high molecular weight keratins (HMWK, 34bE12, Dako); these sections were scored for the presence of cancer in each core on the TMA as described previously [21 (link)–26 (link)]. A single pathologist (LF) scored MUC1 protein staining only in cores in which cancer was present as determined using the H & E and HMWK.
The immunohistochemical staining intensity for MUC1 was defined as absent, weak (faint cytoplasmic staining of scattered cells), moderate (intermediate or heterogeneous cytoplasmic staining in tumor cells), or strong (dense cytoplasmic staining of nearly all tumor cells) as shown in Fig 1.

This study was done on the total of 19 radical prostatectomy specimens obtained from Vancouver Prostate Centre Tissue Bank. The study protocol was approved by University of British Columbia (UBC) Clinical Research Ethics Board (CREB) and Vancouver Coast Hospital Research Institute Research (VCHRI) Research Ethics Board (REB). All patients gave informed consent as approved by UBC CREB and VCHRI REB. Immunohistochemical staining was conducted by Ventana autostainer model Discover XT™ (Ventana Medical System, Tuscan, Arizona) with enzyme‐labeled biotin–streptavidin system and solvent‐resistant DAB Map kit using 1/400 concentration of mouse monoclonal antibody against Cdc25C and 1/800 concentration of goat polyclonal antibody against CLU. All stained slides were digitalized with the SL801 autoloader and Leica SCN400 scanning system (Leica Microsystems, Concord, Ontario, Canada) at magnification equivalent to ×20 and subsequently stored in the SlidePath digital imaging hub (DIH; Leica Microsystems) of the Vancouver Prostate Centre. For each biomarker, representative cores (clearly positive, clearly negative, and mixed positive/negative) were manually identified by a pathologist and values on a four‐point scale were assigned.