The disclosed subject matter relates to an automated determination the stain quality and segmentation quality of a tissue sample. By way of example, separate image data is acquired of an unstained form of a biological specimen, the biological specimen stained with a nuclei marker and the biological specimen stained with a segmentation marker. A correlation map (Cr) from the separate image data and a ridgeness map (Pr) from the image data of the biological specimen stained with a segmentation marker are each determined. A staining quality score and segmentation quality score are then determined from the correlation map (Cr) and the ridgeness map (Pr).