PCK VDS, Tumor Detection PCK VDS, Tumor Detection

Process APP




PCK VDS, Tumor Detection

This APP has been developed to improve quantification of Ki-67, ER and PR positive cells within breast carcinoma, by automatically identifying tumor areas. The APP uses the Virtual Double Staining (VDS) technique, which enables automated and robust detection of tumor regions.

Two serial sections stained for ER and Pancytokeratin (PCK), respectively, must be used in this APP. Tumor regions are identified automatically on the PCK stained slides and the outlined tumor region are overlaid on the Ki-67, ER and PR stained slide, thus automatically identifying tumor regions.

PCK is used as a cytoplasmatic marker to localize the cancer tissue.

In US: For Research Use Only, not for use in diagnostic procedures.


Figure 1

Figure 1

The original TMA-core stained with PCK.

Figure 2

Figure 2

The original TMA-core stained with ER.

Figure 3

Figure 3

Tissue alignment. The PCK and ER TMA-cores are aligned, one on top of the other.

Figure 4

Figure 4

Part of tissue stained for PCK.

Figure 5

Figure 5

The tumor region is segmented in the PCK tissue and outlined with a ROI. The ROI obtained from the PCK tissue is superimposed on the aligned ER tissue (not seen here). The ROI only outlines tumor regions. Tissue that is not tumor and the background are excluded.

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1. American Society of Clinical Oncology/College of American Pathologists Guideline Recommendations for Immunohistochemical Testing of Estrogen and Progesterone Receptors in Breast Cancer, M. E. H. Hammond et al., J. Clin Oncol 28:2784-95, 2010.

2. Learning histopathological patterns, A. Kårsnäs, A. L. Dahl, R. Larsen, Journal of Pathology Informatics, 2(2):12, 2011

3. Segmenting Clustered Nuclei Using H-minima Transform-Based Marker Extraction and Contour Parameterization, C. Jung et al., IEEE Transactions on Biomedical Engineering 2010, 57(10): 2600-2604

Quantitative Output variables

The output consist of one or more ROIs, outlining the tumor areas. The total area of the tumor regions is estimated as the variable ‘Tumor Area’.


To employ the VDS approach for Ki-67, ER and PR, two serial sections must be stained for Ki-67, ER, PR and PCK, respectively.
This APP works as a three-step process as described below.

First, alignment of the two serial sections [See EXAMPLES: figure 3]. The alignment is done both on a large scale, and on a finer detailed level, to get the best possible match of the two tissue sections.

Second, the tumor areas are automatically detected from the PCK slide and outlined as regions of interest (ROIs) [See EXAMPLES: figure 4] and [See EXAMPLES: figure 5]. The ROIs are then superimposed on the aligned ER tissue slide to outline the tumor region for subsequent analysis limited to the inside of the tumor regions.

Staining Protocol

Staining protocols have been developed by the NordiQC and are available from their website. It is also possible to use the links below:


Additional information

This APP has been developed in cooperation with Professor Mogens Vyberg from NordiQC and Aalborg University Hospital Denmark.


VDS, Virtual Double Staining, breast carcinoma, immunohistochemistry, breast cancer, quantitative, digital pathology, image analysis, PCK, Cytokeratin, TMA

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