Fig 3. Photobleaching of BcO binding to AV at 15°C.

The photobleaching rate constant was elucidated with the (A) $\overline{rF}\left(t\right)$ and (B) $\overline{F}\left(t\right)$ signals by collecting the reaction with continuous (black) and discontinuous (dashed color) laser illumination in which the beam was blocked during the times denoted by dashes and the sample was illuminated only during time intervals of 10 s. The slow photobleaching rate constant varied from 6 × 10⁻³ to 1 × 10⁻² s^-1, and was laser power dependent. The (A) $\overline{rF}\left(t\right)$ and (B) $\overline{F}\left(t\right)$ normalization functions were: $\overline{rF}\left(t\right)=\left[rF\left(0\right)-rF\left(t\right)\right]/\left[rF\left(0\right)-rF\left(\mathrm{\infty }\right)\right]$ and $\overline{F}\left(t\right)=\frac{F\left(t\right)}{F\left(0\right)}=\left[F\left(t\right)-rF\left(\mathrm{\infty }\right)\right]/\left[rF\left(0\right)-rF\left(\mathrm{\infty }\right)\right]$, respectively.