I want to measure ROS (H2O2 actually) production on my cells by flow cytometry.
My problem is that every sample should have THE SAME INCUBATION TIME of the probe, since the fluorescence increases over time.
So if I incubate 30 min, and then launch the flow cytometer, and that each reading takes 2min/well on a 96-well plate for example, therefore the last well will be incubated with the probe more time than the first one... and the results are not interpratable. For this reason, i was wondering if someone knows an alternative... I was thinking to fix the cells before starting the reading, so i expect to block the reaction from living cells but i am not sure if it is a recommandable technique and that the fluorescence will be stable over time (over 2h for instance).
Thank you in advance for any suggestion,