(Designed for Fluorescent RNAscope®)
Indica Labs’ Multiplex FISH module quantifies up to four DNA or RNA FISH probes on a cell-by-cell basis. This analysis module can be used with most multiplex RNA FISH assays including RNAscope® and supports the H-score protocol for RNAscope® recommended by the manufacturer, ACD Bio.
In the example above the module measures a two-plex RNAscope assay on pelleted cancer cells. Single cells are identified using the nuclear dye DAPI. The multiplexed red and green probes are measured within the cell boundaries (spots per cell and probe area per cell). The module reports the average number of each probe per cell or across the entire region of analysis. In the analysis markup image, cells positive for the red probe alone are highlighted in different shades of red, cells positive for the green probe alone in different shades of green and cells positive for both probes are highlighted in shades of yellow. The color shading is determined by the number of probes and the corresponding RNAscope H-score, dark color = lower expression and light color = higher expression.