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Giedrius Lelkaitis

Image analysis and virtual double staining in hormone receptor analysis


Hormone receptor status in breast carcinomas is a prognostic and predictive marker, which is performed routinely by estimating percentage tumor cells with positive nuclear IHC reaction for estrogen and progesterone receptors on paraffin-embedded tumor tissue. Additionally, the intensity of nuclear staining may be evaluated. The method is subject to inter-observer variability and estimation (rounding) bias, further influenced by differences in tumor cellularity and overall staining intensity. Image analysis and virtual double staining tries to overcome the aforementioned limitations by combining images of estrogen (nuclear) and pan-cytokeratin (cytoplasmatic) stained consecutive slides of tumor tissue to make an image of virtual double staining, detecting groups or isolated epithelial cells in manually drawn regions of interest, filtering cells without discernible nucleus and counting cells with negative, weak, moderate and strong intensity of estrogen staining.

In the presented study 109 cores of breast carcinomas where stained for estrogen receptor and evaluated using H-score by 5 observers. Image analysis of virtual double staining was performed using the same ER slides and consecutive slides stained for pan-cytokeratin. Result comparison of observer assessment and image analysis will be discussed, regarding inter- and intra-observer variability and estimation bias.


Dr. Giedrius Lelkaitis graduated Medical faculty of Vilnius University in 2001, completed residency of Pathology in National Centre of Pathology at Vilnius University Hospital in 2005. Since 2009 he has been senior consultant at the Institute of Pathology, Aalborg University Hospital with focus on diagnostic breast pathology and head and neck pathology. Dr. Lelkaitis is a board member of Danish Breast Cancer Cooperative group and Danish CancerBiobank Advisory Committee.


Brügmann A, Eld M, Lelkaitis G, Nielsen S, Grunkin M, Hansen JD, Foged NT, Vyberg M. Digital image analysis of membrane connectivity is a robust measure of HER2 immunostains. Breast Cancer Res Treat. 2012 Feb;132(1):41-9. doi: 10.1007/s10549-011-1514-2. Epub 2011 Apr 22. PubMed PMID: 21512768.

Brügmann A, Lelkaitis G, Nielsen S, Jensen KG, Jensen V. Testing HER2 in breast cancer: a comparative study on BRISH, FISH, and IHC. Appl Immunohistochem Mol Morphol. 2011 May;19(3):203-11. doi: 10.1097/PAI.0b013e3181f7118e. PubMed PMID: 21475037.

Mele M, Jensen V, Wronecki A, Lelkaitis G. Myofibroblastoma of the breast: Case report and literature review. Int J Surg Case Rep. 2011;2(6):93-6. doi: 10.1016/j.ijscr.2011.02.006. Epub 2011 Mar 3. PubMed PMID: 22096693; PubMed Central PMCID: PMC3199680.

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