F_norm

For binding affinity analysis, ligand-dependent changes in TRIC are plotted as F_norm values vs. ligand concentration in a dose-response curve. F_norm values are plotted as parts per thousand (‰). For each well (each data point), a TRIC trace is recorded. All traces are then normalized to start at 1 (relative fluorescence). For each trace, the F_norm value for the Dose Response Curve is calculated by dividing F₁ by F₀. F₁ corresponds to the fluorescence value measured in the heated state, while F₀ is the fluorescence value measured in the cold state before the IR laser is turned on. Both values are averages taken between two time points signified by two cursors (red and blue).

Note that an F_norm Outlier can be automatically identified and excluded from analysis by Dianthus software during the fitting of affinity data.