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Abstract
Digitizing slides and analyzing the resulting images with image analysis algorithms has become a standard method to assess staining patterns of tissue processed by immunohistochemistry (IHC). An innovative peptide-based control slide capable of detecting procedural errors that may occur in the analytical phase of the IHC staining process was developed. A software application that quantifies the peptide spots staining intensities and assesses HER2 scoring on cell line and tissue microarrays was developed. Slides processed according to standard operating procedure and by simulating antibody dilution errors were analyzed with this HER2 algorithm. The coefficient of variance (CV) of staining intensity for the peptide spots were less than 1% during the reproducibility assessment. Error detection sensitivity of the peptide-based control was compared with cell line and tissue-based controls. Characteristics that make the developed algorithm robust are presented. These studies demonstrate that images generated by the iScan™ and analyzed by the analysis algorithm are reliable in the quantification of the peptide spot staining intensities on HER2 control slides.