Journal
APPLIED IMMUNOHISTOCHEMISTRY & MOLECULAR MORPHOLOGY
Volume 27, Issue 7, Pages 558-563Publisher
LIPPINCOTT WILLIAMS & WILKINS
DOI: 10.1097/PAI.0000000000000626
Keywords
immunohistochemistry; hematoxylin; eosin; cIQc; counterstain
Funding
- Centre de recherche du Centre Hospitalier de l'Universite de Montreal (CRCHUM)
- Institut du cancer de Montreal (ICM)
- Fonds de Recherche du Quebec-Sante (FRQ-S)
- Mitacs
- Institut du cancer de Montreal
- FRQ-S Clinical Research Scholar, Junior 1
- Rising Star Award (Prostate Cancer Canada)
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Hematoxylin and eosin (H&E) staining is a well-established technique in histopathology. However, immunohistochemistry (IHC) interpretation is done exclusively with hematoxylin counterstaining. Our goal was to investigate the potential of H&E as counterstaining (H&E-IHC) to allow for visualization of a marker while confirming the diagnosis on the same slide. The quality of immunostaining and the fast-technical performance were the main criteria to select the final protocol. We stained multiple diagnostic tissues with class I IHC tests with different subcellular localization markers (anti-CK7, CK20, synaptophysin, CD20, HMB45, and Ki-67) and with double-staining on prostate tissues with anti-high molecular weight keratins/p63 (DAB detection) and p504s (alkaline phosphatase detection). To validate the efficacy of the counterstaining, we stained tissue microarrays from the Canadian Immunohistochemistry Quality Control (cIQc) with class II IHC tests (ER, PR, HER2, and p53 markers). Interobserver and intraobserver concordance was assessed by kappa statistics. Excellent agreement of H&E-IHC interpretation was observed in comparison with standard IHC from our laboratory (kappa, 0.87 to 1.00), and with the cIQc reference values (kappa, 0.81 to 1.00). Interobserver and intraobserver agreement was excellent (kappa, 0.89 to 1.00 and 0.87 to 1.00, respectively). We therefore show for the first time the potential of using H&E counterstaining for IHC interpretation. We recommend the H&E-IHC protocol to enhance diagnostic precision for the clinical workflow and research studies.
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