Resorufin (NSC-12097) (NSC-12097) is a highly fluorescent pink dye.

I. ROS/RNS Detection
1. Prepare solution: Dissolve Resorufin in an appropriate solvent, usually dimethyl sulfoxide (DMSO), to prepare a working solution (e.g., 1-10 μM).
2. Cell incubation: Add Resorufin to cultured cells or tissues and incubate for a set time, usually 30 minutes to 1 hour.
3. ROS/RNS generation: Use stimuli such as H2O2 or NO donors to induce ROS/RNS generation.
4. Fluorescence measurement: Use a fluorescence spectrophotometer, fluorescence microscope, or flow cytometer to measure fluorescence emission at approximately 590 nm at an excitation wavelength of approximately 530 nm.
II. Cellular analysis of oxidative stress
1. Treatment: Add Resorufin to cells after exposure to potential oxidative stressors or antioxidants.
2. Measurement: Measure changes in fluorescence intensity to quantify the level of ROS/RNS in cells.
III. Monitoring of mitochondrial red oxygen reduction potential
1. Mitochondrial incubation: Treat cells with mitochondrial inhibitors or other compounds and add Resorufin to track changes in red oxygen reduction.
2. Fluorescence monitoring: Measure fluorescence changes in real time to observe the dynamics of mitochondrial red oxygen reduction.
IV. Analysis of other analytes:
1. Sample preparation: Prepare biological or chemical samples that may contain analytes of interest.
2. Fluorescence detection: Detect the presence and concentration of specific analytes through fluorescence monitoring.