Qualitative and quantitative evaluation of the tissue micro-environment by high-resolution 17-plex immunofluorescence reveals distinct cell populations
Multiplexed tissue images are only useful if you can both see the cell populations and measure them; this SITC 2021 poster pairs a single-scan 17-plex Orion immunofluorescence panel with HALO AI image analysis to qualitatively visualize and quantitatively profile the distinct cell populations of lymphoid tissue.
Presented by RareCyte and Indica Labs.
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- A single Orion scan captured a 17-plex immunofluorescence panel across whole slides of three lymphoid tissues. FFPE sections of reactive tonsil, quiescent lymph node and SLL/CLL lymphoma were stained with one 17-plex panel — spanning leukocyte, B-cell, T-cell, macrophage, checkpoint, vascular, epithelial and proliferation markers plus a nuclear dye — and imaged whole-slide in a single scan on the Orion instrument, then fed to a downstream HALO AI image-analysis workflow.
- Qualitatively, the images placed distinct cell populations in their spatial context across all three tissues. Follicular B cells (CD20), interfollicular T-cell zones (CD3d, CD4, CD8), macrophages (CD68, CD163), blood vessels (CD31) and proliferating cells (PCNA, Ki-67) were read directly from the panels, showing for instance that nearly all germinal-center B cells in the reactive tonsil were proliferating while the lymphoma cells expressed PCNA but not Ki-67.
- Quantitatively, HALO AI segmented the tissue regions and measured how they differed between the reactive tonsil and quiescent lymph node. The T-cell-to-B-cell zone-area ratio was 27% lower in the tonsil than the lymph node (2.6 versus 3.6); the T-cell region ran 81.9% CD3+ in the lymph node against 50.7% in the tonsil; and the CD4:CD8 ratio fell from 3.1 to 1.4, marking the tonsil’s more balanced helper-to-cytotoxic T-cell mix.
