Measurement and Clinical Significance of Biomarkers of Oxidative Stress in Humans

Table 1

Fluorescent probes used for the measurements of reactive oxygen and nitrogen species by flow cytometry.

Probe (localization)	ROS/RNS	Fluorescence	Leukocytes	Platelets	Limitations and confoundings
DCFH-DA (intracellular)	HO^•; ONOO^−; ROO^•; NO2^•; Indirect; H2O2	↑ green (DCF)	Yes	Yes	Hemolysis; Self-propagation of DCF radicals; MDR substrates or inducers; Esterase inhibitors; Plasma esterase in whole blood or PRP; EDTA and citrate; Antioxidants
DAF-2 DA/DAF-FM DA (intracellular)	NO^•	↑ green (DAF-Ts)	Yes	No	MDR substrates or inducers; Esterase inhibitors; Plasma esterase in whole blood
DHR123 (intracellular)	HClO; H2O2; ONOO^−	↑ green (Rho123)	Yes	No	Self-propagation of DHR radicals; MDR substrates or inducers; Antioxidants
HE (intracellular)	O2^•−	↑ red (ethidium)	Yes	No	Intercalating agents
C11-BODIPY^581/591 (membrane)	HO^•; ROO^•	Shift from red to green	Yes	Yes	Hemolysis; Antioxidants

C11-BODIPY^581/591: 4,4-difluoro-5-(4-phenyl-1,3-butadienyl)-4-bora-3a,4a-diaza-s-indacene-3-undecanoic acid; DAF-2 DA: 4,5-diaminofluorescein diacetate; DAF-FM DA: 4-amino-5-methylamino-2′,7′-difluorofluorescein diacetate; DAF-Ts: triazolofluoresceins; DCF: 2′,7′-dichlorofluorescein; DCFH-DA: dihydrochlorofluorescein diacetate; DHR123: dihydrorhodamine 123; EDTA: ethylenediaminetetraacetic acid, H₂O₂: hydrogen peroxide; HClO: hypochlorous acid; HE: hydroethidine; MDR: multidrug resistance; NO^•: nitrogen monoxide; NO₂^•: nitrogen dioxide; O₂^•−: superoxide radical; HO^•: hydroxyl radical; ONOO⁻: peroxynitrite; PRP: platelet-rich plasma; Rho123: rhodamine 123; ROO^•: peroxyl radicals.