I am planning to do some experiments to detect ROS in arabidopsis and I was wondering what is the difference between using H2DCFDA and Singlet oxygen sensor?
For Singlet oxygen you have to use SOSG, (singlet oxygen sensor green) which is sensitive and give localisation of singlet oxygen only, while DCFDA will give information about total ROS in the cell.
H2DCFDA is a permeable and chemically reduced form of fluorescein, which when enters in the living cells , its acetate groups is cleavage of by intracellular esterases. When these reduced fluoresceins come it contact with singlet, hydroxyl or superoxide radicals, it get oxidised and gives green fluorescence. But SOSG is very less sensitive to hydroxyl or superoxide and give fluorescence only with singlet oxygen species.
For Singlet oxygen you have to use SOSG, (singlet oxygen sensor green) which is sensitive and give localisation of singlet oxygen only, while DCFDA will give information about total ROS in the cell.
H2DCFDA is a permeable and chemically reduced form of fluorescein, which when enters in the living cells , its acetate groups is cleavage of by intracellular esterases. When these reduced fluoresceins come it contact with singlet, hydroxyl or superoxide radicals, it get oxidised and gives green fluorescence. But SOSG is very less sensitive to hydroxyl or superoxide and give fluorescence only with singlet oxygen species.
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