@article{oai:repo.qst.go.jp:00045137, author = {Hafer, Kurt and Konishi, Teruaki and Robert, Schiestl and Hafer Kurt and 小西 輝昭 and Robert Schiestl}, issue = {4}, journal = {Radiation Research}, month = {Apr}, note = {The dichlorofluorescein method has become a standard technique for measuring reactive oxygen species (ROS) formed in cells by ionizing radiation. A recent report [Y.N. Korystov, V.V. Shaposhnikova, A.F. Korystova, M.O. Emel'yanov, Detection of Reactive Oxygen Species Induced by Radiation in Cells Using the Dichlorofluorescein Assay. Radiat. Res. (2007)] has suggested the method is subject to an artifact in which it erroneously reports hydrogen peroxides generated in the extracellular medium as ROS formed intracellularly by ionizing radiation. It was hypothesized that such ionizing radiation-induced extracellular peroxides influx into cells in the minutes following radiation exposure and subsequently oxidize the intracellular dichlorofluorescin probe and that dichlorofluorescein fluorescence is not due to ROS formed by ionizing radiation intracellularly. Here we test this hypothesis by measuring the contribution of long-lived radicals formed in medium by ionizing radiation on intracellular dichlorofluorescein fluorescence. We find no evidence that such an artifact significantly contributes to intracellular dichlorofluorescein fluorescence. These results and those of Y.N. Korystov et al. are discussed in view of cellular dichlorofluorescin leakage and radiation chemistry. We conclude that the dichlorofluorescein method remains effective for quantifying intracellular ROS induced by ionizing radiation.}, pages = {469--473}, title = {Radiation-induced long-lived extracellular radicals do not contribute to measurement of intracellular reactive oxygen species using the dichlorofluorescein method}, volume = {169}, year = {2008} }