Reference standards for histopathological assays are critical for validating, optimizing, and routinely monitoring assay performance in order to maintain quality control and ensure the sensitivity, specificity, and accuracy of your assay. The RNAscope™ ISH assay is a robust technology that identifies RNA expression patterns and localization at the single cell level with morphological context. Here we demonstrate the application of the RNAscope™ assay on the IHC HDx™ Reference Standards from Horizon Discovery. The benefits of this compatibility include:

• Generation of excellent quality RNA ISH staining by the RNAscope™ assay with the IHC HDx™ Reference Standards, validating the use of these cell lines and slides with the RNAscope™ assay
• Generation of staining patterns by the RNAscope™ assay that are highly concordant with protein detection for many established and emerging biomarkers
• Generation of discrete signal patterns by the RNAscope™ assay as opposed to the diffuse signals from IHC, enabling more objective and quantitative assessment of biomarker expression than IHC
• Detection of low expression levels, not detectable by IHC, due to the ultra-sensitive capabilities of the RNAscope™ assay

IHC HDx™ Reference Standards provide confidence through verification and validation activities. The external standard tests the analytical technical performance of your assay with a range of negative and positive protein expressing cell lines and allows to establish the practical lower limit of detection of your assay with validated low protein-expressing cells. The RNAscope™ ISH assay results display a quantitative correlation with the IHC HDx™ Reference
Standards comparable to IHC, which increases confidence and consistency in experimental results. The key benefit to the RNAscope™ strategy is high sensitivity due to its amplification
method and high specificity based on the double Z probe design, resulting in a high signal-to-noise ratio in many tissues.