Detection of irradiation induced reactive oxygen species production in live cells

Bo Gao; Debin Zhu

doi:10.1117/12.710703

27 October 2006 Detection of irradiation induced reactive oxygen species production in live cells

Bo Gao, Debin Zhu

Proceedings Volume 6047, Fourth International Conference on Photonics and Imaging in Biology and Medicine; 604738 (2006) https://doi.org/10.1117/12.710703
Event: Fourth International Conference on Photonics and Imaging in Biology and Medicine, 2005, Tianjin, China

Abstract

Reactive oxygen species (ROS) is thought to play an important role in cell signaling of apoptosis, necrosis, and proliferation. Light irradiation increases mitochondrial reactive oxygen species (ROS) production and mediates its intracellular signaling by adjusting the redox potential in tumor cells. Mitochondria are the main source of ROS in the living cell. Superoxide anions (0₂^- are likely the first ROS generated in the mitochondria following radiation damage, and then convert to hydrogen peroxide (H₂0₂), hydroxyl radical (•OH), and singlet oxygen (¹0₂), etc. Conventional methods for research ROS production in mitochondria mostly use isolated mitochondria rather than mitochondria in living cells. In this study, a highly selective probe to detect mitochondrial 0₂^- in live cells, MitoSOX^TM Red, was applied to quantify the mitochondrial ROS production in human lung adenocarcinoma cells (ASTC-a-1) with laser scanning microscope (LSM) after ultraviolet C (UVC) and He-Ne laser irradiation. Dichiorodihydrofluoresein diacetate (DCFHDA), a common used fluorescent probe for ROS detection without specificity, were used as a comparison to image the ROS production. The fluorescent image of MItoSOX^TM Red counterstained with MitoTracker Deep Red 633, a mitochondria selective probe, shows that the mitochondrial ROS production increases distinctly after UVC and He-Ne laser irradiation. DCFH-DA diffuses labeling throughout the cell though its fluorescence increases markedly too. In conclusion, the fluorescent method with MitoSOX^TM Red reagent is proved to be a promising technique to research the role of ROS in radiation induced apoptosis.

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Bo Gao and Debin Zhu "Detection of irradiation induced reactive oxygen species production in live cells", Proc. SPIE 6047, Fourth International Conference on Photonics and Imaging in Biology and Medicine, 604738 (27 October 2006); https://doi.org/10.1117/12.710703

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KEYWORDS

Oxygen

Helium neon lasers

Luminescence

Cell death

Laser irradiation

Microscopes

Ultraviolet radiation

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