The Influence of Staining Procedures on the Assessment of Cell Proliferation as Defined by the Monoclonal Antibody KI-67

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Abstract
We examined the influence of different staining techniques [(three-step immunoperoxidase technique (IP): alkaline phosphatase–anti–alkaline phosphatase technique (APAAP)] on the quantitative evaluation of Ki-67–labeled nuclei. We studied five melanocytic skin tumors. From each case, five parallel sections were prepared and stained using the peroxidase-antiperoxidase (PAP) technique (slide 1) and the APAAP technique once (slide 2). Slide 3 consisted of a single repetition of the APAAP technique, slide 4 was a double repetition, and slide 5 was a third repetition. We assessed the volume fraction (VV) of Ki-67–positive nuclei using computer-assisted image analysis. For each staining group, the mean value and standard deviation of VV were calculated. Comparing VV values obtained from the different staining groups we did not find a statistically significant difference between the IP and the various APAAP steps (Wilcoxon test, p =

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