Unsupervised method for normalization of hematoxylin-eosin stain in histological images

•We present a normalization method of H&E-stained histological images.•To do so, histological and H&E properties were considered.•Public images datasets were used for performance comparison.•Visual and quantitative analysis prove the relevance of histological normalization.•The influence of highly eosin stained regions demands future researches. Histological images stained with hematoxylin-eosin are widely used by pathologists for cancer diagnosis. However, these images can have color variations that highly influence the histological image processing techniques. To deal with this potential limitation, normalization methods are useful for color correction. In this paper, a histological image color normalization is presented by considering the biological and hematoxylin-eosin properties. To this end, the stain representation of a reference image was applied in place of the original images representation, allowing the preservation of histological structures. This proposal was evaluated on histological images with great variations of contrast, and both visual and quantitative analyzes yielded promising results.