Third, validated IHC assays may be implemented readily into clinical practice. Second, clinical laboratories typically perform IHC on FFPE tissue sections processed by standard methods, making potentially available hundreds of millions of specimens for study. This is an important advantage over “grind and bind” assays in which tissue is solubilized for biochemical analysis, which may lead to false negative results if few biomarker-positive cells are present in a background of biomarker-negative tissue elements. First, it allows direct visualization of biomarker expression in histologically relevant regions of the examined tissue. Immunohistochemistry (IHC) is an important technique for biomarker validation for several reasons.
The virtual slide(s) for this article can be found here: This study demonstrated that computer-aided methods to classify image areas of interest (e.g., carcinomatous areas of tissue specimens) and quantify IHC staining intensity within those areas can produce highly similar data to visual evaluation by a pathologist.
Image pro plus vs image j software#
Comparisons of IHC intensity data derived using pixel analysis software versus pathologist visual scoring demonstrated high Spearman correlations of 0.88 for %Pos (p < 0.0001) and 0.90 for OD*%Pos (p < 0.0001).
Image pro plus vs image j manual#
A comparison of the IHC staining data obtained from manual annotations and software-derived annotations showed strong agreement, indicating that software efficiently classifies carcinomatous areas within IHC slide images. Two metrics for IHC staining were used: the percentage of carcinoma with S100A1 staining (%Pos), and the product of the staining intensity (optical density of staining) multiplied by the percentage of carcinoma with S100A1 staining (OD*%Pos). To evaluate computer-aided image classification, IHC staining within pathologist annotated and software-classified areas of carcinoma were compared for each case. Using TMAs representing 215 ovarian serous carcinoma specimens stained for S100A1, we assessed the degree to which data obtained using computer-aided methods correlated with data obtained by pathologist visual scoring. Computer-aided analysis of digitized whole slide images may overcome these limitations. However, the method of pathologist semi-quantification is costly, inherently subjective, and produces ordinal rather than continuous variable data. Due to the ubiquitous availability of IHC techniques in clinical laboratories, validated IHC biomarkers may be translated readily into clinical use. IHC is useful for validating biomarkers discovered through genomics methods as large clinical repositories of FFPE specimens support the construction of tissue microarrays (TMAs) for high throughput studies.
Immunohistochemical (IHC) assays performed on formalin-fixed paraffin-embedded (FFPE) tissue sections traditionally have been semi-quantified by pathologist visual scoring of staining.
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