ImmunoMembrane: a publicly available web application for digital image analysis of HER2 immunohistochemistry

• Published in: Histopathology Vol. 60; no. 5; pp. 758 - 767
• Main Authors: Tuominen, Vilppu J, Tolonen, Teemu T, Isola, Jorma
• Format: Journal Article
• Language: English
• Published: Oxford, UK Blackwell Publishing Ltd 01.04.2012; Blackwell; Wiley Subscription Services, Inc
• Subjects: Algorithms; Biological and medical sciences; Biomarkers, Tumor - metabolism; breast cancer; Breast Neoplasms - diagnosis; Breast Neoplasms - metabolism; Chromogenic Compounds; DNA, Neoplasm - analysis; Female; Genetic Markers; Gynecology. Andrology. Obstetrics; human epidermal growth factor receptor 2; Humans; Image Processing, Computer-Assisted; immunohistochemistry; Immunohistochemistry - methods; In Situ Hybridization, Fluorescence; Investigative techniques, diagnostic techniques (general aspects); Mammary gland diseases; Medical sciences; open source software; Pathology. Cytology. Biochemistry. Spectrometry. Miscellaneous investigative techniques; quantification; Receptor, ErbB-2 - genetics; Receptor, ErbB-2 - metabolism; Reproducibility of Results; Software; Tissue Array Analysis; Tumors; Immunohistochemistry; Breast disease; quantification; open source software; erbB2 Gene; Quantization; Breast cancer; Malignant tumor; Digital image; Human Epidermal growth factor Receptor 2; Mammary gland diseases; Image analysis; Anatomic pathology; C-Onc gene; Software; Internet; Protooncogene; Cancer
• ISSN: 0309-0167; 1365-2559; 1365-2559
• DOI: 10.1111/j.1365-2559.2011.04142.x

Tuominen V J, Tolonen T T & Isola J 
(2012) Histopathology 60, 758–767
ImmunoMembrane: a publicly available web application for digital image analysis of HER2 immunohistochemistry Aims:  Assessment of the human epidermal growth factor receptor 2 (HER2) with immunohistochemistry (IHC) is routine practice in clinical pathology laboratories. Visual classification of the staining reaction (usually into 0/1+, 2+ or 3+) is subjective and prone to significant inter‐ and intra‐observer variation. In this study, we describe ImmunoMembrane, an easy‐to‐use HER2 IHC analysis software, which is freely available as a web application, requiring no download or installation. Methods and results:  ImmunoMembrane uses colour deconvolution for stain separation and a customized algorithm for cell membrane segmentation. A quantitative score (IM‐score, 0–20 points) is generated according to the membrane staining intensity and completeness. Specimens are classified into 0/1+, 2+ or 3+ based on IM‐score cut‐offs defined using a training set. The classification and membrane segmentation are presented as a pseudo‐coloured overlay image. With a validation set (144 HercepTest®‐stained whole tissue sections), ImmunoMembrane matched well with the pathologist’s visual classification (weighted kappa κw = 0.80), as well as fluorescence in‐situ hybridization (FISH) (IHC disagreement 3.5%, n = 144) and chromogenic in‐situ hybridization (CISH) (IHC disagreement 2.8%, n = 144). Conclusions:  We anticipate that publicly available web applications, such as ImmunoMembrane, will accelerate the adoption of automated image analysis in clinical diagnostics of HER2 IHC. ImmunoMembrane is freely accessible at: http://jvsmicroscope.uta.fi/immunomembrane/.