Exploring interplay between tumor intrinsic features and immune cell phenotypes in multi-tumor tissue microarrays by a single step 15-plex fluorescence immunohistochemistry and imaging on the Orion Platform
Immunohistochemistry in clinical practice reads one to six markers and needs multiple rounds of antigen retrieval over two days to get them; this AACR 2024 poster from Navigate BioPharma stained 15 markers on a single FFPE slide in one step, imaged them on the Orion platform, and checked the result against the laboratory’s own clinical-grade assays.
Presented by Navigate BioPharma Services.
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- The single-step 15-plex panel tracked the laboratory’s own clinical-grade assays across 11 shared biomarkers. Linear regression on 14 tissue microarray cores returned R² from 0.72 (CD68) to 0.98 (CK), and slopes from 0.69 to 1.10, against serial sections stained with the in-house 6-plex TSA-OPAL workflow and imaged on PhenoImager HT — agreement with an existing assay across those cores, not a standalone measure of accuracy.
- One staining step stood in for multiple rounds of antigen retrieval spread over two days. The stated problem is that immunohistochemistry in clinical practice is restricted to one to six markers and relies on sequential staining over two days, carrying a high risk of tissue and signal loss; here all 15 markers were applied in a single step to one FFPE slide, which could then be processed for H&E.
- Immune phenotypes were read cell by cell in place, beside the tumor cells they neighbor. The rare-cell panels show an exhausted regulatory T cell carrying four markers at once (CD3e+CD4+LAG3+FOXP3+); CD68+CD163+ M2 macrophages sitting next to PDL1+ tumor cells; and a proliferating cytotoxic T cell (CD3+CD8a+Ki67+) beside proliferating CK+Ki67+ tumor cells, the last two both in head and neck carcinoma.
