Orion™ Applications

Imaging in up to 21 markers across an entire slide in a single scan enables spatial analysis though examination of biomarker expression, cell classification, and subcellular localization studies to advance pathology, immuno-oncology, infectious disease, and more.

Visualize micro-anatomy

B-cell follicles with germinal centers and interfollicular T-cell zones define the lymphoid micro-anatomy of the tonsil where immune response is generated to upper respiratory tract antigens. Displayed markers: SYTOX blue (nucleus, gray); CD45 (leukocytes, blue); CD3e (T cells, green); CD20 (B cells, red).

Investigate T-cell subsets

Helper, cytotoxic, memory, regulatory and activated/exhausted T cells are simultaneously revealed. Displayed markers: SYTOX blue (nucleus, gray); CD4 (helper T cells, green); CD8a (cytotoxic T cells, red); CD45RO (memory T cells, blue); LAG-3 (activated/exhausted T cells, pink); FoxP3 (T-reg cells, magenta).

Characterize tissue architecture

Cancer tissue micro-environments contain numerous cell types, including epithelial, immune, vascular endothelial and smooth muscle cells in a spatial context for immune response. Displayed markers: SYTOX blue (nucleus, gray); E-cadherin (mucosal and crypt epithelium, magenta); pan-cytokeratin (mucosal and crypt epithelium, cyan) smooth muscle actin (arterioles, red); CD31 (vascular endothelium and crypt sub-mucosa, green); and CD20 (B cells, orange); CD3e (T cells, blue).

Examine checkpoint biomarkers

PD-1/PD-L1 expression in tumor cells and the tumor micro-environment impacts response to checkpoint inhibitor therapy. Displayed markers: SYTOX blue (nucleus, gray); PD-1 (germinal centers, yellow); PD-L1 (crypt sub-mucosa, green); CD20 (B cells, blue); CD3e (T cells, magenta).

21-marker imaging of lung carcinoma with the Orion Instrument

A single lung section was stained with TissuePlex reagents and imaged using the Orion Instrument. All 21 markers were stained in a single-staining procedure and imaged in a single scan. Three regions of interest (ROIs) are shown below that highlight the 21 markers. Additional single channel images are shown in grayscale for clarity.

Neoplastic Lung

Image above shows cytokeratin-positive, focally TTF1-positive carcinoma in ROI #1. CD8a (magenta), Nuclei (blue), Pan-cytokeratin (green), SMA (white), and TTF1 (red).

Merge	CD31	CD45	CD11b

GFAP	CD20	FOXP3	PD-L1

ROI #1. Single channel images demonstrate blood vessels (CD31), immune cell markers (CD45, CD11b, CD20, FOXP3), and checkpoint biomarker (PD-L1).

Nonneoplastic Lung

Image above shows area of non-neoplastic lung tissue in ROI #2. CD8a (magenta), Nuclei (blue), Pan-cytokeratin (green), and TTF1 (red).

Merge	Pan-CK	SMA	CD163

E-Cadherin	TTF1	CD4	LAG3

ROI #2. Single channel images demonstrate lung epithelial cells (Pan-CK, E-Cadherin, TTF1), blood vessels (SMA), and immune cell markers (CD163, CD4, and LAG3).

Lymphoid Aggregate

Lymphoid aggregate is shown above with germinal center in ROI #3. PD-1 (purple), CD8a (magenta), CD3d (red), CD11b (green), and Ki67 (blue).

Merge	Ki67	CD11c	CD68

CD3d	CD8a	PD-1	Nuclei

ROI #3. Single channel images demonstrate proliferation (Ki67), immune cell markers (CD11c, CD68, CD3, and CD8a), and the checkpoint biomarker (PD-1).