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Rasmus Røge

Virtual double staining in the assessment of proliferation markers


Ki67 is a proliferation marker important for classification of malignant tumours such as breast carcinoma. As part of the NordiQC international immunohistochemical external quality assessment scheme, 370 laboratories were invited to score proliferation indices on twenty Ki67 stained breast carcinomas on a virtual slide using their preferred method. Results varied greatly underlining the necesity of standardization of interpretation of Ki67 stained slides. Digital image analysis has been suggested as the crux of modern pathology, simplying laborous and uniform tasks such as calculation of proliferation indices. Visiopharm A/S (Denmark) recently introduced a new Virtual Double Staining technique where two parallel sections stained for different markers can be virtually fused combining the information in both images. Using this technique, two parallel sections (from the same tumours as used in the webmodule) stained for either Pancytokeratin or Ki67 were combined and allowed the program to calculate a Ki67 proliferation index for tumour cells only. Obtained proliferation indices were compared to manually assessed proliferation indices, where more than 500 cells from each core were examined by experienced pathologists for Ki67 positivity. The results will be presented at the conference.


Resident in Pathology, Aalborg University Hospital. In addition working as scheme organizer and assessor in the Nordic Immunohistochemical Quality Control (NordiQC) General and Breast Cancer module.

Fields of Research: Quality Assurance in Immunohistochemistry, Proliferation Markers, Digital Image Analysis.

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