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Hofmann-Wellenhof, R; Smolle, J; Kerl, H.
The influence of staining procedures on the assessment of cell proliferation as defined by the monoclonal antibody Ki-67.
Am J Dermatopathol. 1990; 12(5):458-461 Doi: 10.1097/00000372-199010000-00005
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Leading authors Med Uni Graz: Hofmann-Wellenhof Rainer; Kerl Helmut
Co-authors Med Uni Graz: Smolle Josef
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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 = less than 0.05). However, the staining procedure influenced the quantitative results to some extent. The mean VV of the five staining groups ranged in our study from 0.10 to 0.17%, which is narrow compared with the overall variability among different cases (dermal melanocytic nevus, 0.01%; metastatic malignant melanoma, 0.43%). Therefore, we can state that for a rough evaluation of Ki-67-positive nuclei, the influence of different staining methods is negligible; for a subtle quantitative analysis, however, it would nevertheless be preferable to always apply the same staining technique.
Find related publications in this database (using NLM MeSH Indexing): Antigens, Surface - analysis; Cell Division - analysis; Humans - analysis; Image Interpretation, Computer-Assisted - methods; Immunoenzyme Techniques - methods; Ki-67 Antigen - methods; Melanocytes - pathology; Melanoma - metabolism; Nevus - metabolism; Nuclear Proteins - analysis; Skin Neoplasms - metabolism; Staining and Labeling - metabolism