Multiplex Immunofluorescence Staining, Whole Slide Imaging, and Spatial Phenotyping of T cell Exhaustion, Regulatory T cells, and Myeloid Derived Suppressor Cells in Tumor FFPE Samples
Profiling exhausted T cells, regulatory T cells, and myeloid-derived suppressor cells in the tumor microenvironment demands many markers, spatial context, and fast turnaround at once; this SITC 2019 poster combines three UltiMapper I/O immunofluorescence panels, whole-slide scanning on the RareCyte CyteFinder II HT, and HALO image analysis to phenotype these populations across lung, colorectal, and breast tumor FFPE samples with same-day sample-to-results.
Presented by Indica Labs / RareCyte / Ultivue.
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- Three UltiMapper I/O multiplex immunofluorescence panels profiled the tumor immune microenvironment across lung, colorectal, and breast FFPE tissue. Each panel was imaged in 5 channels — a DAPI channel plus four markers — with a PD-1 panel for immune-cell exhaustion, a T-reg panel for regulatory and cytotoxic T cells, and an MDSC panel for myeloid-derived suppressor cells.
- The automated workflow returned same-day sample-to-results, from staining through analysis. Staining on a Leica BOND RX autostainer completed in 5.5 hours, whole-slide scanning ran about 15 minutes per slide on the RareCyte CyteFinder II HT, and HALO image analysis finished in 10 minutes.
- Serial sections stained with the three panels were co-registered in HALO onto one coordinate system for spatial phenotyping. Cytotoxic T-cell infiltration was quantified 200 µm inside and 200 µm outside the tumor margin, and T-helper-cell density was measured within a 25 µm radius, resolving immune phenotypes by their position relative to the tumor margin and stroma.
