Multispectral imaging: a review of its technical aspects and applications in anatomic pathology

Vet Pathol. 2014 Jan;51(1):185-210. doi: 10.1177/0300985813506918. Epub 2013 Oct 15.


The field of anatomic pathology has changed significantly over the last decades and, as a result of the technological developments in molecular pathology and genetics, has had increasing pressures put on it to become quantitative and to provide more information about protein expression on a cellular level in tissue sections. Multispectral imaging (MSI) has a long history as an advanced imaging modality and has been used for over a decade now in pathology to improve quantitative accuracy, enable the analysis of multicolor immunohistochemistry, and drastically reduce the impact of contrast-robbing tissue autofluorescence common in formalin-fixed, paraffin-embedded tissues. When combined with advanced software for the automated segmentation of different tissue morphologies (eg, tumor vs stroma) and cellular and subcellular segmentation, MSI can enable the per-cell quantitation of many markers simultaneously. This article covers the role that MSI has played in anatomic pathology in the analysis of formalin-fixed, paraffin-embedded tissue sections, discusses the technological aspects of why MSI has been adopted, and provides a review of the literature of the application of MSI in anatomic pathology.

Keywords: immunofluorescence; multicolor immunohistochemistry; multispectral imaging; pathology; per-cell quantitation; tissue cytometry.

Publication types

  • Review

MeSH terms

  • Animals
  • Biomarkers, Tumor / analysis*
  • Biomarkers, Tumor / metabolism
  • Fluorescent Antibody Technique / veterinary*
  • Humans
  • Image Processing, Computer-Assisted
  • Immunohistochemistry / veterinary*
  • Paraffin Embedding / veterinary
  • Pathology, Molecular / methods*
  • Pathology, Veterinary / methods*
  • Software
  • Tissue Fixation / veterinary


  • Biomarkers, Tumor