Simultaneous, high-throughput detection of RNA and protein biomarkers using the Orion™ imaging system and HCR™ Gold for comprehensive analysis of tumor samples in clinical trials
Multiomic tissue analysis usually means cyclic staining that can take days and degrade the sample; this AACR 2025 proof-of-concept poster from RareCyte and Molecular Instruments detects RNA and protein biomarkers on the same FFPE tumor section, pairing single-round Orion protein imaging with a 1-step, protease-free HCR Gold RNA-ISH workflow.
Presented by RareCyte.
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- One 15-plex assay reads RNA and protein on the same FFPE tissue section. 10 protein markers stained with ArgoFluor-conjugated antibodies, 4 RNA markers detected by HCR Gold, and a nuclear dye were combined on a single whole-slide section, with CD8a read as both an RNA transcript and a protein.
- A 1-step, protease-free RNA protocol and single-round protein staining replace the multi-day cycles of cyclic multiplexing. HCR Gold amplifies RNA signal without protease or enzyme digestion, while Orion scans up to 18 biomarkers in one staining round, avoiding the tissue degradation and days-to-weeks processing of cyclic staining methods.
- The proof-of-concept panel resolved tumor-infiltrating immune cells across non-small cell lung, gastric and colon tumors. Whole-tissue images captured cytotoxic and helper T cells, macrophages, B cells and memory T cells, and pinpointed individual T cells expressing cytokine RNA — IFNγ and TNFα — infiltrating the tumor, read alongside CD8a and MALAT1 transcripts on the same sections.
