High-Quality Immunohistochemical Stains through Computational Assay Parameter Optimization

Abstract

Accurate profiling of tumors using immunohistochemistry (IHC) is essential in cancer diagnosis. The inferences drawn from IHC-stained images depend to a great extent on the quality of immunostaining, which is in turn affected strongly by assay parameters. To optimize assay parameters, the available tissue sample is often limited. Moreover, with current practices in pathology, exploring the entire assay parameter space is not feasible. Thus, the evaluation of IHC stained slides is conventionally a subjective task, in which diagnoses are commonly drawn on images that are suboptimal. In this work, we introduce a framework to analyze IHC staining quality and its sensitivity to process parameters. To that extent, first histopathological sections are segmented automatically. Then, machine learning techniques are employed to extract disease-specific staining quality metrics (SQMs) targeting a quantitative assessment of staining quality. Finally, an approach to efficiently analyze the parameter space is introduced to infer sensitivity to process parameters. We present results on microscale IHC tissue samples of five breast tumor classes, based on disease state and protein expression. A disease-type classification F1-score of 0.82 and a contrast-level classification F1-score of 0.95 were achieved. With the proposed SQMs, an area under the curve of 0.85 was achieved on average over different disease types. Our methodology provides a promising step in automatically evaluating and quantifying staining quality of IHC stained tissue sections, and it can potentially standardize immunostaining across diagnostic laboratories.