A method to reuse archived H&E stained histology slides for a multiplex protein biomarker analysis

James P. Hinton, Katerina Dvorak, Esteban Roberts, Wendy J. French, Jon C. Grubbs, Anne E. Cress, Hina A. Tiwari, Raymond B. Nagle

Research output: Contribution to journalArticlepeer-review

16 Scopus citations


Archived Hematoxylin and Eosin (H&E) stained pathology slides are routinely stored to index formalin-fixed paraffin-embedded (FFPE) sample tissue blocks. FFPE blocks are clinically annotated human tumor specimens that can be valuable in studies decades after the tissue is collected. If stored properly, they have the potential to yield a valuable number of serial sectioned slides for diagnostic or research purposes. However, some retrospective studies are limited in scope because the tissue samples have been depleted or not enough material is available in stored blocks for serial sections. The goal of these studies was to determine if archived H&E-stained slides can be directly reutilized by optimizing methods to de-stain and then re-stain the H&E stained slides to allow the detection of several biomarkers of interest using a conjugated antibody with chromogen multiplex immunohistochemistry procedure. This simple but innovative procedure, combined with image analysis techniques, demonstrates the ability to perform precise detection of relevant markers correlated to disease progression in initially identified tumor regions in tissue. This may add clinical value in retaining H&E slides for further use.

Original languageEnglish (US)
Article number86
Pages (from-to)1-15
Number of pages15
JournalMethods and Protocols
Issue number4
StatePublished - Dec 2019


  • De-stain
  • H&E stained slides
  • Image analysis
  • Immunohistochemistry
  • Re-stain

ASJC Scopus subject areas

  • Biotechnology
  • Structural Biology
  • Biochemistry, Genetics and Molecular Biology (miscellaneous)


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