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Superoxide anion (O 2 •-) is an important biomarker for reactive oxygen species (ROS) generated through physiological and pathological processes. However, due to the short half-life of O 2 •- and high autofluorescence of cell tissues, in situ real-time tracking and monitoring of endogenous O 2 •- can be difficult. In this paper, a fluorescent probe IFP-O 2 was developed to detect endogenous O 2 •- in cells. The probe could instantaneously react with O 2 •- to produce fluorescence off-on effect; its detection limit was as low as 10 nM. Cell experiments also showed that the probe had low toxicity and mitochondrial targeting ability. The article presents, for the first time, a probe that can be employed to measure endogenous O 2 •- in oral cancer Cal-27 cells and is a promising tool for monitoring and evaluating apoptosis. Image 1 [ABSTRACT FROM AUTHOR]