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After digital stain separation of original chromogenic IHC images, quantification of the diaminobenzidine (DAB) reaction pattern was performed based on intensity and extent of cytoplasmic, nuclear, and stromal expression.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>Assessment of molecular expression proposed that c\u2010Myc and HIF\u20101\u03b1 may be used as strong screening markers, VEGF for risk\u2010stratification and VEGFRII and CD105 for prognosis of precancer into oral cancer.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion<\/jats:title><jats:p>Our analysis indicated that the proposed method can help in establishing IHC as an effective quantitative immunoassay for molecular pathology and alleviate diagnostic ambiguities in the clinical decision process. \u00a9 2015 Wiley Periodicals, Inc. 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