Background Tissue consists of heterogenous cell types, each with diverse functions and functional states, arranged spatially in a way that impacts patient health status. Resolving this complexity at the subcellular level has historically been challenged by fluorescence overlap, autofluorescence, a limited number of targets that can be simultaneously assessed, and low throughput. Orion technology breaks these barriers, providing rapid, straightforward, highly multiplexed whole slide tissue analysis. The recently announced Orion platform rapidly generates high resolution 21-channel images to enable comprehensive phenotypic profiling and characterization of tissue architecture including micro-anatomy, analysis of tumor heterogeneity and the complex tissue microenvironment.

Methods FFPE tissue samples are stained with TissuePlex™ reagents and whole slides are imaged on the Orion Instrument. Resulting 21-channel datasets are reviewed by a pathologist to verify staining specificity. The TissuePlex core panel includes antibodies against CD45, CD4, FoxP3, CD8A, CD11b, LAG-3, PD-L1, CD11c, CD163, CD68, PD-1, Ki-67, CD3d, and CD20. Single marker IHC is used as the gold standard to evaluate TissuePlex reagents and instrument spectral isolation performance is evaluated using fluorescent microspheres.

Results We will present the novel Orion spatial biology platform, provide a technology and workflow overview, demonstrate instrument validation results, and present sample datasets.

Conclusions The Orion platform enables rapid and deep phenotypic analysis of tissue samples for high resolution, whole slide sample analysis.