详细信息:Millipore’s FlowCellect™ MitoLive Kit includes MitoSense Red, a fluorescent cationic dye that accumulates in the mitochondria and is responsive to mitochondrial potential changes and Calcein acetoxymethylester (calcein-AM) a non-fluorescent, cell permeant compound that is hydrolyzed by intracellular esterases into the fluorescent anion calcein and provides a measure of cellular vitality. The simultaneous use of the reagents allows researchers to obtain information on early and late apoptosis in one simple assay. MitoSense Red is excitable by a red laser and fluoresces maximally at 650 nm (Red2 channel fluorescence on the guava easyCyte 8HT). Uninduced cells with intact mitochondrial membrane potential demonstrate high Red2 fluorescence while cells which have impaired mitochondrial membrane potential depict lower Red2 fluorescence. The cell permeant dye, Calcein-AM included in the kit, enters all cells and if the cell is vital is hydrolyzed by intracellular esterases into the fluorescent anion calcein which are retained in cells with intact membranes. Healthy cells thus exhibit high green fluorescence. Cells in which the intracellular esterases are impacted on treatment or have compromised membranes demonstrate low green fluorescence. The assay kit allows the identification of 3 populations of cells; healthy live cells with intact membrane potential, live cells with impaired mitochondria and non-vital or dead cells with impaired mitochondria. The entire assay can be performed in approximately 30 min once cellular samples are ready.
