Is there any ways to measure ROS without using FACS? I'd like toi know the easiest way of measuring ROS.
There are cell permeable fluorescent and chemiluminescent probes. 2'-7'-Dichlorodihydrofluorescein diacetate (DCFH-DA) is one of the most widely used techniques for directly measuring the redox state of a cell. The cell-permeant H2DCFDA passively diffuses into cells and is retained in the intracellular level after cleavage by intracellular esterases. Upon oxidation by ROS, the nonfluorescent H2DCFDA is converted to the highly fluorescent 2',7'-dichlorofluorescein (DCF). you can use this for intracellular detection of ROS by FACS. All the best!!
I agree with Mansoor Hussain, DCF is really usefull but you have to keep in mind that this method just mensures peroxyl radical and don't all of kind of ROS.
Could you let me know the condition that detect DCF well in fresh kidney tissues?
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