We have measured the ROS contents in algal cells after bacterial extracts treatment, and the method as followed:
Intracellular ROS were detected using a fluorescent probe,
2′,7′-dichlorofluorescin diacetate (DCFH-DA), based on
Choudhury et al. (2013), with slight modifications. The final
DCFH-DA concentration in the mixture was 10 μM and this
was incubated with the suspended cells at 37 °C in the dark for
30 min and mixed every 5min during this time. Then, the cells
were immediately washed three times with sterile f/2 medium
(without silicate) and finally suspended with 500 μL f/2
medium. The fluorescence intensity was monitored using a
spectrofluorometer with excitation wavelength at 488 nm and
emission wavelength at 525 nm.
hope this answer could be helpful to you!
Dear Yi Li, thank you for the kind response and methodology for measuring the ROS. I perform the same in bacterial cells.
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