Orion™: 16-plex single-step stain and imaging of oral squamous cell carcinoma

Highly multiplexed tissue imaging usually builds its picture over repeated rounds of staining and imaging; this SITC 2023 poster shows a whole FFPE section of oral squamous cell carcinoma stained with a 16-plex panel in one round and imaged on Orion in a single scan.

Presented by RareCyte.

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• One staining round, one imaging round, on a standard FFPE section. The section was stained in one round with a 16-plex immunofluorescence panel, using ArgoFluor-conjugated antibodies and standard IF protocols, imaged whole-slide at 20X on the Orion instrument in a single scan with tissue autofluorescence isolated as an additional channel, and written out as OME-TIFF; the protocol then de-coverslips and H&E-stains the same section.
• A single scan separates normal tongue mucosa, invasive tumor and the stroma between them. E-Cadherin identifies the basal epithelial layer and most tumor cells while Pan-CK marks squamous differentiation; Ki67 shows the tumor to be far more proliferative than the normal epithelium; and CD31 capillary endothelium, SMA and MYH11 in arteriolar walls, FAP in fibroblasts, and Vimentin across many stromal cell types resolve the submucosal stroma.
• The immune response is read cell by cell at the tumor border. CD20 B cells, CD4 helper T cells and CD8a cytotoxic T cells gather within an active area of cellular fibrosis marked by FAP at the edge of the tumor nodule; in a T cell aggregate beside nests of tumor cells, FOXP3 identifies regulatory T cells and the checkpoint marker PD-1 marks activated cells, including lone CD8 T cells that strongly express PD-1 within the tumor nests.