Microchip electrophoresis with laser-induced fluorescence detection for the determination of the ratio of nitric oxide to superoxide production in macrophages during inflammation

In this investigation, ME–LIF was used for the simultaneous detection of nitric oxide and superoxide using the fluorescent probes DAF-FMDA and MitoSOX Red. This method was also used to study the variations of the nitric oxide/superoxide ratio in RAW 264.7 macrophage cell lysates under physiological and proinflammatory conditions. Additionally, the effect of the natural antioxidant carnosine and the second messenger calcium in modulating this ratio was investigated. These results highlight the roles played by different stimulation protocols in influencing the release and bioavailability of nitric oxide with respect to superoxide.