A Quantitative Method to Measure Low Levels of ROS in Nonphagocytic Cells by Using a Chemiluminescent Imaging System

Figure 2

Development of the chemiluminescent imaging method by using luminol-based chemiluminescence to measure low levels of ROS. Various concentrations of H₂O₂ (0, 1, 5, and 10 μM) were mixed with luminol (50 μM in DMSO or NaOH) with or without 4IPBA (180 μM) in a black-walled glass bottom 96-well plate. HRP (0.2 U/ml) were added and chemiluminescence was assessed by signal accumulation mode (SAM, every 30 sec for 300 sec) using a chemiluminescent imaging system. (a) Representative chemiluminescent images. (b) Densitometry analyses of chemiluminescent images were plotted (, ±SD). : linear correlation coefficient.

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