Scenario description
In this scenario, multiple target expression patterns in one or more cell types are being investigated (Figures 21, 22 and 23).
Figure 21. Multiplex expression of KRT19 (green), PECAM1 (red) and MKI67 (white) in human breast cancer using the RNAscope ® 2.5 LS Multiplex Fluorescent Assay. Cells are counterstained with DAPI (blue).
Figure 22. Multiplex expression pattern for Vglut1 (red), Vglut2 (green) and Vgat (white) in mouse brain using the RNAscope ® 2.5 LS Multiplex Fluorescent Assay. Cells are counterstained with DAPI (blue).
Figure 23. Multiplex expression of TGFβ1 (green), CD8A (yellow), CD4 (red) and TNFα (white) in human lung cancer using the RNAscope ® 2.5 LS Multiplex Fluorescent Assay. Cells are counterstained with DAPI (blue).
Analysis guidelines
One can qualitatively and quantitatively assess the expression pattern by using methodologies #1, #2 and #3. For further guidelines on how to quantify RNAscope ® fluorescent assay results see Tech Note SOP 45-006.
To understand co-localization across markers and positivity in specific cell populations, percentage of cells positive can be scored visually at 40x based on number of cells with ≥1 dot/cell.
