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Multiplex hyperspectral IHC: A 12-plex assay for high-dimensional analysis of the tumor microenvironment.

Created on 16 Oct 2026

Authors

Kouther Noureddine, Paul Gallagher, John English, Jagoda Korbelik, Anita Carraro, Sylvia Lam, Puneet Arora, Athena Li, Martial Guillaud, Anna McGuire, Calum MacAulay

Published in

ImmunoHorizons. Volume 10. Issue 10. Oct 15, 2026.

Abstract

The immune component of the tumor microenvironment (TME) is highly prognostic of patient outcome, further understanding of TME and the spatial organization of the immune cells within the TME is needed for better patient prognosis and treatment planning. Current brightfield immunohistochemistry techniques quantify immune cell counts and density but generally cannot assess the spatial relationship between tumor and immune cells due to the limited number of markers on a single slide. Here, we present a brightfield-based multiplexed immunohistochemistry workflow that combines multiple labels per staining round with iterative cycles of staining, destaining, and hyperspectral imaging to enable high-dimensional immune profiling within a single tissue section. This approach is directly compared with a sequential immunofluorescence technique applied to adjacent tissue sections to compare and evaluate its performance. By integrating serial imaging, sequential labeling, and image registration, we are able to spatially map the TME using standard brightfield microscopy.

PMID:
42885232
Bibliographic data and abstract were imported from PubMed on 16 Oct 2026.

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