Figure 16.

It is impossible to regroup blinking events belonging to a single fluorophore using thresholds when signals from different fluorophores overlap or superpose. For illustrative purposes only, we depict the signal from one hypothetical PA-FP in orange and another in blue. Although methods based on grouping events using a blinking correction time cannot identify which event belongs to which fluorophore in such cases, this does not preclude them from yielding an accurate count.