I have carried out the DCF-DA assay to determine the intra-cellular ROS production caused by the r-GO in fungus candida albicans and got green fluorescence. Which ROS species is responsible for this phenomina. is it H2O2 or OH. radical? To determine the production of any extracellular OH radical, I have carried out terphthalic acid ROS scavenger test. But the result was negative. Then what will be the possible mechanism and source of this intra-cellular ROS production?.
You valuable comments are highly appreciated.....
DCF is not a specific ROS probe. It can be converted by various cellular components that are not strictly related to oxidative stress. Please refer to the attached papers for more details. For example, Amplex Red or MitoSOX are better ROS probes.
Regards, Christian
Article What does the commonly used DCF test for oxidative stress really show?
Article Measurement of Reactive Oxygen Species, Reactive Nitrogen Sp...
Dear Prof. Christian Q. Scheckhuber
Thank you for your reply. Is the situation is same for both eukaryotic and prokaryotic cells?
Dear Mahesh,
I would assume that the situation is the same in prokaryotic cells because they also contain compounds that can convert DCF like heme enzymes and metal ions.
Cheers, Christian
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