(Designed for RNAscope®)
Indica Labs FISH-IF quantification module measures any number of fluorescently-labeled DNA/RNA ISH probes and immunofluorescent (IF) protein biomarkers on a cell-by-cell basis. This allows the user to rapidly contextualize the corresponding protein and gene expression profile of every cell across the tissue. HALO FISH-IF analysis is designed to work with single or dual IHC-ISH assays including RNAscope™ and supports the H-score protocol for RNAscope as recommended by the manufacturer, ACD Bio.
An example of FISH-IF quantification is shown in the slider above. Here, DAPI and an IF-labeled membrane biomarker (cyan) are used to identify cells of interest and the RNA FISH probe (pink) is measured in these cells. Cells that express the membrane biomaker are highlighted in cyan. The FISH probe spots are highlighted in pink. IF stains can be identified and quantified in any compartment – nucleus, cytoplasm and/or membrane.
The module reports numerous outputs including, but not limited to: number of IF positive cells in total and by cell compartment, Average IF intensity and average IF intensity by cell compartment, percentage IF positive total and by cell compartment, number and percentage of probe positive cells, probe copies per cell and area per cell, cell score for each probe (0, +1, +2, +3 and +4), and an H-score for each probe. Additionally, users can define cell phenotypes based on IF and probe positivity.