Color normalization of faded H&E-stained histological images using spectral matching

Abstract

Histological samples stained with hematoxylin-eosin (H&E) are commonly used by pathologists in cancer diagnoses. However, the preparation, digitization, and storage of tissue samples can lead to color variations that produce poor performance when using histological image processing techniques. Thus, normalization methods have been proposed to adjust the color of the image. This can be achieved through the use of a spectral matching technique, where it is first necessary to estimate the H&E representation and the stain concentration in the image pixels by means of the RGB model. This study presents an estimation method for H&E stain representation for the normalization of faded histological samples. This application has been explored only to a limited extent in the literature, but has the capacity to expand the use of faded samples. To achieve this, the normalized images must have a coherent color representation of the H&E stain with no introduction of noise, which was realized by applying the methodology described in this proposal. The estimation method presented here aims to normalize histological samples with different degrees of fading using a combination of fuzzy theory and the Cuckoo search algorithm, and dictionary learning with an initialization method for optimization. In visual and quantitative comparisons of estimates of H&E stain representation from the literature, our proposed method achieved very good results, with a high feature similarity between the original and normalized images.