Deep phenotyping and spatial interrogation of the NSCLC tumor microenvironment using the fully validated Next Generation MultiOmyx™ Platform
Multiplexed immunofluorescence is often limited to a small imaging area; this SITC 2024 NeoGenomics poster demonstrates a Next Generation MultiOmyx (MO 2.0) workflow that generates whole-tissue data from a 16-marker panel across 20 NSCLC samples on the CyteFinder II microscope, benchmarked against clinically validated IHC and the prior MO 1.0 workflow.
Presented by Indica Labs.
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- The MO 2.0 workflow images a 16-marker panel across whole NSCLC tissue sections, rather than only a selected region of interest. The immunofluorescence panel spans immune, stroma, and tumor biomarkers and is applied over 8 iterative staining rounds — a Cy3 and a Cy5 marker each round — on the CyteFinder II microscope, using a customized software package co-developed by RareCyte and NeoGenomics to drive whole-tissue acquisition across a set of 20 NSCLC samples.
- Marker readouts from the MO 2.0 workflow were checked against two independent references. Tumor H-scores for Ki67 and PD-L1 measured by MO 2.0 immunofluorescence matched clinically validated IHC on serial sections with strong concordance (r = 0.94-0.97), while Pearson correlation of cell density (count/area) for all 16 panel biomarkers between the earlier MO 1.0 and the MO 2.0 workflows averaged r = 0.96.
- Repeatability held across separate runs, with every biomarker inside the preset variability limit. 3 NSCLC samples were stained in triplicate across 3 separate batches — 9 serial sections in total — and inter-run and intra-run coefficients of variation were calculated for both cell density and staining intensity, with all study biomarkers meeting the acceptability criterion of %CV at or below 25%.
