Extended Data Fig. 3. Qualifying 16-plex single-shot Orion antibody panel relative to immunohistochemistry and Cyclic Immunofluorescence (CyCIF).
a, Panels of images from FFPE tonsil sections showing single-antibody immunohistochemistry (IHC) for the indicated markers and matching channels extracted from the 16-plex Orion immunofluorescence (IF) images (H&E stain was performed on the same section as the Orion imaging). Scalebars 50 µm. Images are from one representative tonsil specimen. b, Plots of the fraction of positive for the indicated markers (CD45, CD68, CD20, CD4, FOXP3) from whole-slide Orion IF and CyCIF images acquired from two adjacent sections of 29 FFPE colorectal cancer specimens. Pearson correlation coefficients are indicated. c, t-distributed stochastic neighbor embedding (t-SNE) plots of cells from Orion IF image. Log transformed marker intensities (CD31, CD20, E-cadherin, Ki-67) were used to color the dots in each panel. Fig. 2d contains tSNE plots for additional markers and the inferred cell types. Single cell data is from one representative specimen (C01).