Validation of tumor protein marker quantification by two independent automated immunofluorescence image analysis platforms.

摘要: Protein marker levels in formalin-fixed, paraffin-embedded tissue sections traditionally have been assayed by chromogenic immunohistochemistry and evaluated visually pathologists. Pathologist scoring of chromogen staining intensity is subjective generates low-resolution ordinal or nominal data rather than continuous data. Emerging digital pathology platforms now allow quantification fluorescence signals computer-assisted image analysis, providing values. Fluorescence offers greater dynamic signal range immunohistochemistry, combined with analysis holds the promise enhanced sensitivity analytic resolution, consequently more robust quantification. However, commercial scanners software differ features capabilities, claims objective quantitative are difficult to validate as pathologist there no accepted gold standard. Here we provide first side-by-side validation two technologically distinct platforms. We document highly consistent results (1) concordance values (2) agreement outcome predictions both for objective, data-driven cutpoint dichotomization Kaplan-Meier analyses employment compute receiver-operating curves. The examined rely on imaging hardware, microscopy vs line scanning, functionally software. nuclear-localized tyrosine-phosphorylated Stat5a/b computed each platform a cohort 323 breast cancer cases revealed high after linear calibration, finding confirmed an independent 382 case cohort, correlation coefficients >0.98. Data-driven optimal cutpoints prediction either were reciprocally applicable derived alternate platform, identifying patients low Nuc-pYStat5 at ~3.5-fold increased risk disease progression. Our identified concordant that may serve benchmarks testing other platforms, interoperator variability supports implementation tumor laboratories.