Table 18.4 Fluorescence response of 3′-(p-aminophenyl) fluorescein (APF), 3′-(p-hydroxyphenyl) fluorescein (HPF) and dichlorodihydrofluorescein diacetate (H2DCFDA) to various reactive oxygen species (ROS).
| Reactive Oxygen Species (ROS) | ROS Generation Method | APF * | HPF * | H2DCFDA * |
|---|---|---|---|---|
| Hydrogen peroxide (H2O2) | 100 µM of H2O2 | <1 | 2 | 190 |
| Hydroxyl radical (HO•) | 100 µM of ferrous perchlorate (II) and 1 mM of H2O2 | 1200 | 730 | 7400 |
| Hypochlorite anion (–OCl) | 3 µM (final) of –OCl | 3600 | 6 | 86 |
| Nitric oxide (NO) | 100 µM of 1-hydroxy-2-oxo-3-(3-aminopropyl)-3-methyl-1-triazene (NOC-7) | <1 | 6 | 150 |
| Peroxyl radical (ROO•) | 100 µM of 2,2'-azobis(2-amidinopropane), dihydrochloride (AAPH) | 2 | 17 | 710 |
| Peroxynitrite anion (ONOO–) | 3 µM (final) of ONOO– | 560 | 120 | 6600 |
| Singlet oxygen (1O2) | 100 µM of 3-(1,4-dihydro-1,4-epidioxy-1-naphthyl)propionic acid | 9 | 5 | 26 |
| Superoxide anion (•O2–) | 100 µM KO2 | 6 | 8 | 67 |
| Autooxidation | 2.5 hours exposure to fluorescent light source | <1 | <1 | 2000 |
* 10 µm of APF, HPF or DCF (2',7'-dichlorofluorescein) were added to sodium phosphate buffer (0.1 M, pH 7.4); ROS were generated as indicated; and fluorescence was measured using excitation/emission wavelengths of 490/515 nm (for APF and HPF) or 500/520 nm (for DCF). DCF was obtained by hydrolysis of H2DCFDA with base as described in  ; dihydrofluorescein diacetates are colorless and nonfluorescent until both of the acetate groups are hydrolyzed and the products are subsequently oxidized to fluorescein derivatives. | ||||
For Research Use Only. Not for use in diagnostic procedures.
