title: ImmunoMembrane: a publicly available web application for digital image analysis of HER2 immunohistochemistry.
abstract: OBJECTIVE
  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.
  
  
  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 κ(w) =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/.
authors.affiliation: Institute of Biomedical Technology, University of Tampere, Tampere, Finland.
authors.forename: Vilppu J
authors.initials: VJ
authors.lastname: Tuominen
authors.forename: Teemu T
authors.initials: TT
authors.lastname: Tolonen
authors.forename: Jorma
authors.initials: J
authors.lastname: Isola
mesh_terms: D000465:Algorithms; D014408:Biomarkers, Tumor / Q000378:metabolism; D001943:Breast Neoplasms / Q000175:diagnosis* / Q000378:metabolism; D002863:Chromogenic Compounds; D004273:DNA, Neoplasm / Q000032:analysis; D005260:Female; D005819:Genetic Markers; D006801:Humans; D007091:Image Processing, Computer-Assisted; D007150:Immunohistochemistry / Q000379:methods*; D017404:In Situ Hybridization, Fluorescence; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000235:genetics / Q000378:metabolism*; D015203:Reproducibility of Results; D012984:Software; D046888:Tissue Array Analysis
chemical_list: D014408:Biomarkers, Tumor; D002863:Chromogenic Compounds; D004273:DNA, Neoplasm; D005819:Genetic Markers; D018719:Erb-b2 Receptor Tyrosine Kinases; C508053:ERBB2 protein, human