I have done previously ROS by DHE stain using flowcytometry. My study design included adherent cell culture: Cells after seeding, treatment, trypsinizing, pellet was incubated with FBS-free medium + DHE dye for 15 min in dark at room temperature. Then centrifuged and washed with chilled PBS, and read using PBS in flowcytometer.

Now i want to try using fluorescent microplate reader.. any reliable protocol that I can follow?

What kind of buffer i should use to read results?

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