Automated IHC Quantification of Whole Tissue and TMA Slides

Digital images of both IHC-stained whole tissue sections and IHC-stained TMA slides were captured using a whole slide scanner at 10× magnification (Aperio Scanscope, Leica Microsystems, Milton Keynes, UK), and the digital images were stored in SVS format. The digital images were retrieved using a file management web interface (Aperio eSlide Manager, version 12.4.3.8007, Leica Biosystems, IL, USA) and reviewed with the server software (Aperio Imagescope, version 12.3.3.5048, Leica Biosystems, IL, USA). Digital images of representative whole tissue section fields and TMA cores were extracted through the server software (Aperio Imagescope, version 12.3.3.5048, Leica Biosystems, IL, USA) at 10× magnification and quantified into H-scores using an automated IHC profiler built in a digital image analysis software (ImageJ version 1.52a, National Institute of Health, Bethesda, MD, USA) [50 (link)].