I did ROS assay using H2-DCFDA but could not find the function of detecting 2 wavelengths ( ex: 485, em: 520) at the same time as single reading in microplate reader so I got readings separately at the above mentioned wavelengths. I am not familiar well with this wavelength thing so I don't know how to analyze my result as I did ROS assay first time and don't even have idea that how it looks as combined reading. Can anyone please help me how to analyze the data? Any formula to calculate ROS from these wavelength values?

I have 2 separate readings (480nm and 520nm) for ROS assay.

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