Well, I found a paper pretty old, 2001, that shows a decrease in the DCFH-DA signal. Is this decrease a quenching effect? Or another phenomenon but never a measurement of antioxidant enzymatic.
What do you think?
help me a little bit whit this, OP
The intracellular redox machinery, including antioxidant enzymes, can indeed influence the oxidation of dichloro-dyhydro fluorescein diacetate (DCFH-DA) within cells. DCFH-DA is commonly used as a probe to measure cellular oxidative stress levels.
When DCFH-DA enters cells, it is deacetylated by intracellular esterases to form dichloro-dyhydro fluorescein (DCFH). DCFH can then react with reactive oxygen species (ROS) within the cells, resulting in the formation of the fluorescent compound 2',7'-dichlorofluorescein (DCF). This fluorescence can be measured and used as an indicator of intracellular oxidative stress.
However, it's important to note that the decrease in DCFH-DA signal observed in the paper you mentioned may not necessarily be a quenching effect. Other factors could contribute to the decrease, such as the enzymatic breakdown of DCFH-DA or alterations in the intracellular redox environment.
To accurately determine the specific phenomenon responsible for the decrease in the DCFH-DA signal observed in the study, further investigations and experiments are needed. These could include measuring the activity of antioxidant enzymes, such as superoxide dismutase, catalase, or glutathione peroxidase, as well as evaluating the effect of ROS scavengers or modulators on the DCFH-DA signal.
It's worth noting that since the paper you mentioned is from 2001, subsequent research might have provided more insights into the mechanism of DCFH-DA oxidation and the interpretation of changes in the DCFH-DA signal. Consulting more recent literature on the topic can help provide a more comprehensive understanding of the current knowledge and experimental approaches in this field.
Thanks a lot. According to my experience, It is pretty unlikely that redox enzymes do something to the fluorescein molecule; enzymes are specific, and they are not reducing around any molecule.
As you noted, the paper is old. By now, nobody dares to say that fluorescein derivatives are suitable for observing intracellular reductions, but the result caught my attention.
Cheers, OP
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