Use of diaminofluoresceins to detect and measure nitric oxide in low level generating human immune cells

Nitric oxide (
- NO) has been implicated in multiple physiological and pathological immune processes. Different methods have been developed to detect and quantify
- NO, where one of the principal difficulties are the accurately detection in cellular system with low levels of
- NO production. The choice of the
- NO detection method to be used depends on the characteristics of the experimental system and the levels of
- NO production which depend on either the organism source of samples or the experimental conditions. Recently, high sensitive methods to detect and image
- NO have been reported using 4,5-diaminofluorescein-based fluorescent probes (DAF) and its derivate 4,5-diaminofluorescein diacetate (DAF-2 DA). This work was aimed to adapt and optimize the use of DAF probes to detect and quantify the
- NO production in systems of high, moderate and low out-put production, especially in human PBMC and their subpopulations. Here, we report an original experimental design which is useful to detect and estimate
- NO fluxes in human PBMC and their subpopulations with high specificity and sensitivity.