Characteristics of microvasculature can be revealed by immunohistochemical tissue staining, but manual quantification of these characteristics on whole-slide images containing potentially hundreds of vessels on each is tedious and subject to operator variability. Conventionally, manual quantification is performed for select regions of the whole tissue section where the regions may not be representative of the pathology. Major challenges to automated segmentation are the irregularity of staining

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... of vessel walls, resulting in apparent fragmentation of intact vessels on the images, and artefactual appearance of DAB stain on structures other than vessel walls. Our objective was to develop and validate a fully automated segmentation of the vascular smooth muscle layer on whole-section histology of wild type and regenerated post-ischemia mouse hind limb microvasculature, stained for smooth muscle using 3,3'-Diaminobenzidine (DAB) immunostain.