The Cytonuclear IHC module measures cell-by-cell IHC positivity for single or multi stain applications. Each cell is measured for cytoplasmic or nuclear positivity of every individual stain as well as the colocalization of different marker combinations. Positive cells are binned and assigned an intensity score (0, 1+, 2+, and 3+) based on stain optical density. The module also reports a variety of other metrics relating to cell/nuclear size and stain intensity. Optionally, the tool can output the complete table of cell data including individual cell sizes and optical density measurements for each stain in each cellular compartment. This module has the option to define specific cell phenotypes according to marker positivity, allowing users to define and count specific cell populations based off of defined positivity and negativity rules.
In the first example shown above, the module is configured to output an intensity markup for a single IHC marker (DAB). Cells below the positivity threshold for the DAB marker are blue and cells above the positivity threshold are scoring according to optical density thresholds set in the module, weak positive (1+) in yellow, moderate positive (2+) in orange and strong positive (3+) in red.
In the second example, the module is used to analyze a dual-stain IHC and a co-expression mark-up is generated. Cells positive for the red cytoplasmic marker only are red, cells positive for the brown cytoplasmic marker are green and dual positive cells are cyan.