Cytonuclear FL measures the immunofluorescent positivity of up to three markers in the nucleus or cytoplasm and reports the number of cells that are positive for each marker, the intensity score for each cell (0, 1+, 2+, 3+), and colocalization of different marker combinations. Optionally, the tool can output the complete table of cell data including individual cell sizes and intensity measurements for each dye in each cellular compartment.
In the dual stained mouse pancreas shown above, the cytonuclear FL module is used to 1) identify cells in mouse pancreas positive for one or both biomarkers and 2) score intensity of the nuclear biomarker.
In the first set of images, a co-expression markup is generated where negative cells are marked blue in the nucleus, cells positive for the cytoplasmic biomarker are marked red in the cytoplasm, cells positive for the green biomarker are marked green in the nucleus and dual positive cells are marked both red and green.
In the second set of images, an intensity markup is generated. Here, the nuclear intensity of the green biomarker only is evaluated. Nuclei that are negative for the green biomarker are blue, yellow indicates weak positivity (1+), orange indicates moderate positivity (2+), and red indicates strong positivity (3+).