Preparation of the dyes:
PBFI-AM (Molecular Probes) 50 ug is dissolved in 8.5 ul of DMSO and 8.5 ul of Pluronic F-127. Working concentration 2.5 mM
SBFI-AM (Molecular Probes) 50 ug is dissolved in 8.9 ul of DMSO and 8.9 ul of Pluronic F-127. Working concentration 2.5 mM
Corona Green (Molecular Probes) 50 ug is dissolved in 15.25 ul of DMSO and 15.25 ul of Pluronic F-127. Working concentration of 2.5 mM
Loading of the dyes:
Add 2 ul of PBFI-AM or SBFI AM or Corona Green per 1 ml of cells (approximately 5 x 15 cells per ml) for a final concentration of 5 uM and vortex gently.
Incubate cells are at 37°C for 60 minutes to load the dye.
Immediately prior of flow cytometric examination, add 1 ul of propidium iodide (PI; 10 mg/ml stock) for a final concentration of 10 ug/ml.