Slides were prepared for a representative section of each of the 373 formalin-fixed and paraffin-embedded (FFPE) tissue samples; FFPE tissue section (4-µm-thick) were prepared and stained with H&E using an automatic stainer for routine diagnostic purposes. Representative slides containing deeply invasive tumor parts were selected to visually assess the TSR and tumor pT stage.
For multiplexed H&E- and IHC-stained sections, IHC staining for cytokeratin (CK) (Novocastra™ Liquid Mouse Monoclonal Antibody with 1:500 dilution) was performed. Intermediate filaments found in epithelial cells of all types and markers for carcinoma cells were first analyzed using BOND-MAX Autoimmunostainer (Leica Biosystems, Melbourne, Australia). The IHC-stained slides were scanned at × 200 total magnification (tissue-level pixel size, 0.32 μm/pixel) with Aperio Digital Pathology Slide Scanner (Aperio Technologies, Inc., Vista, CA). After scanning, the same slides were stained with H&E after removing aminoethylcarbazole (AEC) by washing with distilled water, and then, the slides were incubated with 70%, 80%, and 95% ethanol for 2 min each, followed by incubation with 0.15 M KMnO4 for 1 min as previously described20 (link). A total of 15 pairs of multiplexed H&E and CK whole-slide images (WSIs) of gastric carcinoma were prepared for this study.
For multiplexed H&E- and IHC-stained sections, IHC staining for cytokeratin (CK) (Novocastra™ Liquid Mouse Monoclonal Antibody with 1:500 dilution) was performed. Intermediate filaments found in epithelial cells of all types and markers for carcinoma cells were first analyzed using BOND-MAX Autoimmunostainer (Leica Biosystems, Melbourne, Australia). The IHC-stained slides were scanned at × 200 total magnification (tissue-level pixel size, 0.32 μm/pixel) with Aperio Digital Pathology Slide Scanner (Aperio Technologies, Inc., Vista, CA). After scanning, the same slides were stained with H&E after removing aminoethylcarbazole (AEC) by washing with distilled water, and then, the slides were incubated with 70%, 80%, and 95% ethanol for 2 min each, followed by incubation with 0.15 M KMnO4 for 1 min as previously described20 (link). A total of 15 pairs of multiplexed H&E and CK whole-slide images (WSIs) of gastric carcinoma were prepared for this study.
Free full text: Click here
