||Gleason grading is the most widely used system for determining the severity of prostate cancer. The Gleason grade is determined visually under a microscope from prostate tissue that is most often stained with Hematoxylin-Eosin (H&E). In an earlier study we demonstrated that this stain is not ideal for machine learning applications, but that other stains, such as Sirius-hematoxylin (Sir-Htx), may perform better. In this paper we illustrate the advantages of this stain over H&E for blind color
decomposition. When compared to ground truth defined by an experienced pathologist, the relative root-mean-square errors of the color decomposition mixing matrices for Sir-Htx are better than those for H&E by a factor of two, and the Pearson correlation coefficients of the density maps resulting from the decomposition of Sir-Htx-stained tissue gives a 99% correlation with the ground truth. Qualitative examples of the density maps confirm the quantitative findings and illustrate that the density maps will allow accurate segmentation of morphological features that determine the Gleason grade.