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 inFig 1
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
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