Color Normalization in Breast Cancer Immunohistochemistry Images Based on Sparse Stain Separation and Self-Sparse Fuzzy Clustering

Diagnostics (Basel). 2025 Sep 12;15(18):2316. doi: 10.3390/diagnostics15182316.

Abstract

Background and Objective: The color normalization of breast cancer immunohistochemistry (IHC)-stained images helps change the color distribution of undesirable IHC-stained images to be more interpretable for the pathologists. This will affect the Allred score that the pathologists use to estimate the drug quantity for treating breast cancer patients. Methods: A new color normalization technique based on sparse stain separation and self-sparse fuzzy clustering is proposed. Results: The quaternion structural similarity was used to measure the quality of the normalization algorithm. Our technique has a structural similarity score lower than other techniques, and the color distribution similarity is closer to the target. We applied automated and unsupervised nuclei classification with Automatic Color Deconvolution (ACD) to test the color features extracted from normalized images. Conclusions: The classification result from our unsupervised nuclei classification with ACD is similar to other normalization methods, but it offers an easier perception to the pathologists.

Keywords: breast cancer; color deconvolution; fuzzy clustering; histopathological images; immunohistochemistry.