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- Deep phenotyping and spatial interrogation of the NSCLC tumor microenvironment using the fully validated Next Generation MultiOmyx™ Platform

# 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.

Read or download to see:

- 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%.

[Download poster](https://cdn.sanity.io/files/s3p6uqps/production/4d6bfdd654c94f14375dfcd41aa3494bb7569e4b.pdf)

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