title: Registration of whole immunohistochemical slide images: an efficient way to characterize biomarker colocalization.
abstract: OBJECTIVE
  Extracting accurate information from complex biological processes involved in diseases, such as cancers, requires the simultaneous targeting of multiple proteins and locating their respective expression in tissue samples. This information can be collected by imaging and registering adjacent sections from the same tissue sample and stained by immunohistochemistry (IHC). Registration accuracy should be on the scale of a few cells to enable protein colocalization to be assessed.
  
  
  METHODS
  We propose a simple and efficient method based on the open-source elastix framework to register virtual slides of adjacent sections from the same tissue sample. We characterize registration accuracies for different types of tissue and IHC staining.
  
  
  RESULTS
  Our results indicate that this technique is suitable for the evaluation of the colocalization of biomarkers on the scale of a few cells. We also show that using this technique in conjunction with a sequential IHC labeling and erasing technique offers improved registration accuracies.
  
  
  CONCLUSIONS
  Brightfield IHC enables to address the problem of large series of tissue samples, which are usually required in clinical research. However, this approach, which is simple at the tissue processing level, requires challenging image analysis processes, such as accurate registration, to view and extract the protein colocalization information.
  
  
  CONCLUSIONS
  The method proposed in this work enables accurate registration (on the scale of a few cells) of virtual slides of adjacent tissue sections on which the expression of different proteins is evidenced by standard IHC. Furthermore, combining our method with a sequential labeling and erasing technique enables cell-scale colocalization.
authors.affiliation: Laboratories of Image, Signal Processing and Acoustics, École Polytechnique de Bruxelles, Université Libre de Bruxelles, Bruxelles, Belgium DIAPath, Center for Microscopy and Molecular Imaging (CMMI), Gosselies, Belgium.
authors.forename: Xavier
authors.initials: X
authors.lastname: Moles Lopez
authors.affiliation: DIAPath, Center for Microscopy and Molecular Imaging (CMMI), Gosselies, Belgium.
authors.forename: Paul
authors.initials: P
authors.lastname: Barbot
authors.affiliation: DIAPath, Center for Microscopy and Molecular Imaging (CMMI), Gosselies, Belgium.
authors.forename: Yves-Rémi
authors.initials: YR
authors.lastname: Van Eycke
authors.affiliation: Department of Pathology, Erasme Hospital, Bruxelles, Belgium.
authors.forename: Laurine
authors.initials: L
authors.lastname: Verset
authors.affiliation: Department of Pathology, Erasme Hospital, Bruxelles, Belgium.
authors.forename: Anne-Laure
authors.initials: AL
authors.lastname: Trépant
authors.affiliation: DIAPath, Center for Microscopy and Molecular Imaging (CMMI), Gosselies, Belgium.
authors.forename: Lionel
authors.initials: L
authors.lastname: Larbanoix
authors.affiliation: DIAPath, Center for Microscopy and Molecular Imaging (CMMI), Gosselies, Belgium Department of Pathology, Erasme Hospital, Bruxelles, Belgium.
authors.forename: Isabelle
authors.initials: I
authors.lastname: Salmon
authors.affiliation: Laboratories of Image, Signal Processing and Acoustics, École Polytechnique de Bruxelles, Université Libre de Bruxelles, Bruxelles, Belgium DIAPath, Center for Microscopy and Molecular Imaging (CMMI), Gosselies, Belgium.
authors.forename: Christine
authors.initials: C
authors.lastname: Decaestecker
mesh_terms: D015415:Biomarkers / Q000032:analysis*; D002477:Cells / Q000737:chemistry*; D006652:Histological Techniques; D007090:Image Interpretation, Computer-Assisted; D007150:Immunohistochemistry
chemical_list: D015415:Biomarkers
keywords: Biomarker; Colocalization; Digital pathology; Image registration; Immunohistochemistry; Whole slide imaging