Quantitative kinetics of intracellular singlet oxygen generation using a fluorescence probe

• Published in: Scientific reports Vol. 10; no. 1; p. 10616
• Main Authors: Murotomi, Kazutoshi, Umeno, Aya, Sugino, Sakiko, Yoshida, Yasukazu
• Format: Journal Article
• Language: English
• Published: England Nature Publishing Group 30.06.2020; Nature Publishing Group UK
• Subjects: 3T3 Cells; Animals; Fluorescence; Fluorescent Dyes; Fluorescent indicators; Hep G2 Cells; Humans; Intracellular; Mammalian cells; Mice; Mitochondria; Mitochondria - metabolism; Oxidation; Oxidation-Reduction; Physiology; Reactive oxygen species; Singlet Oxygen - metabolism; Time-Lapse Imaging
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/s41598-020-67155-7

Singlet oxygen ( O ) is a type of reactive oxygen species involved in numerous physiological activities. We previously reported that O -specific oxidation products are increased in patients with prediabetes, suggesting that measurement of O may be an important indicator of physiological and pathological conditions. The turnover in the generation and quenching of O is extremely rapid during biological activities owing to it high reactivity and short lifetime in solution. However, the dynamic changes in O generation in living cells have not been fully explored. In this study, we investigated whether the kinetics of O generation can be quantified using a far-red fluorescent probe for mitochondrial O , Si-DMA, following addition of the O generator, endoperoxide, to mammalian cells. The kinetics of Si-DMA fluorescence intensity dose-dependently increased following treatment of mammalian living cells with endoperoxide. Alternatively, treatment with O quenchers decreased the fluorescence intensities following endoperoxide treatment. Our results indicate that the kinetics of intracellular O can be readily obtained using Si-DMA and time-lapse imaging, which provides new insights into the mechanism of O generation in mammalian cells and the exploration of O generators and quenchers.