In this work, we present a new algorithm and benchmark dataset for stain separation in histology images. Histology is a critical and ubiquitous task in medical practice and research, serving as a gold standard of diagnosis for many diseases. Automating routine histology analysis tasks could reduce health care costs and improve diagnostic accuracy. One challenge in automation is that histology slides vary in their stain intensity and color; we therefore seek a digital method to normalize the appearance of histology images. As histology slides often have multiple stains on them that must be normalized independently, stain separation must occur before normalization. We propose a new digital stain separation method for the universally-used hematoxylin and eosin stain; this method improves on the state-of-the-art by adjusting the contrast of its eosin-only estimate and including a notion of stain interaction. To validate this method, we have collected a new benchmark dataset via chemical destaining containing ground truth images for stain separation, which we release publicly. Our experiments show that our method achieves more accurate stain separation than two comparison methods and that this improvement in separation accuracy leads to improved normalization.

We propose a new mathematical and algorithmic framework for unsupervised image segmentation, which is a critical step in a wide variety of image processing applications. We have found that most existing segmentation methods are not successful on histopathology images, which prompted us to investigate segmentation of a broader class of images, namely those without clear edges between the regions to be segmented. We model these images as occlusions of random images, which we call textures, and show that local histograms are a useful tool for segmenting them. Based on our theoretical results, we describe a flexible segmentation framework that draws on existing work on nonnegative matrix factorization and image deconvolution. Results on synthetic texture mosaics and real histology images show the promise of the method.