Background: Digital image (DI) analysis avoids visual subjectivity in interpreting\nimmunohistochemical stains and provides more reproducible results. An automated\nprocedure consisting of two variant methods for quantifying the cytokeratin-19\n(CK19) marker in breast cancer tissues is presented.\nMethods: The first method (A) excludes the holes inside selected CK19 stained areas,\nand the second (B) includes them. 93 DIs scanned from complete cylinders of tissue\nmicroarrays were evaluated visually by two pathologists and by the automated\nprocedures.\nResults and conclusions: There was good concordance between the two\nautomated methods, both of which tended to identify a smaller CK19-positive area\nthan did the pathologists. The results obtained with method B were more similar to\nthose of the pathologists; probably because it takes into account the entire positive\ntumoural area, including the holes. However, the pathologists overestimated the\npositive area of CK19. Further studies are needed to confirm the utility of this\nautomated procedure in prognostic studies.
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