Fluorescence of Dyes in Solutions with High Absorbance. Inner Filter Effect Correction
Figure 4
The dependence of the total fluorescence intensity of ATTO-425 on the solution absorbance.
On Panels A and B the red curves F(AFL) and F0(AFL) represent the dependences of the total fluorescence intensity on the solution absorbance calculated according to Eq. 2 and Eq. 3 (q = 0.9), respectively. The dependence of the ATTO-425 fluorescence on its absorbance recorded by a Cary Eclipse spectrofluorimeter is given on Panel A (black curve); this dependence after correction for the primary inner filter effect and for the primary and secondary inner filter effects is given in Panel B (curves 1 and 2, respectively). For comparison, the dependence of the total fluorescence on absorbance for NATA is given by the green curve (q = 0.14 [21]). Insets: Fluorescence spectra of ATTO-425 in solutions with different absorbance values (black curve 1–0.1, 2; dark red curve 2–0.2, dark green curve 3–0.5, dark yellow curve 4–0.7, dark blue curve 5–0.9, dark violet curve 6–2.8, dark blue curve 7–5.7, pink curve 8–9.5, red curve 9–13.9, green curve 10–20.1, blue curve 11–29.4): recorded (Panel A) and corrected for primary inner filter effect (Panel B).