In this study, a novel approach to measure the absolute cytoplasmic Caconcentration([Ca]cyt) using the Caindicator fluo-3 AM was established. The parameters associated with the probe fluo-3 AM were optimized to accurately determine fluorescence intensity from the Ca-bound probe. Using three optimized parameters(final concentration of 6 m M probe,incubation time of 135 min, loading probe before plasma treatment), the maximum fluorescence intensity(F?=?527.8 a.u.) and the minimum fluorescence intensity(F?=?63.8 a.u.) were obtained in a saturated Casolution or a solution of lacking Ca. Correspondingly, the maximum [Ca]cytinduced by cold plasma was 1232.5 n M. Therefore, the Caindicator fluo-3 AM was successfully applied to measure the absolute [Ca]cytin Saccharomyces cerevisiae stimulated by cold plasma at atmospheric air pressure.
