Gao Jing Jing, Xu Ke Hua, Tang Bo, Yin Ling Ling, Yang Gui Wen, An Li Guo
College of Chemistry, Chemical Engineering and Materials Science, Shandong Normal University, Jinan, China.
FEBS J. 2007 Apr;274(7):1725-33. doi: 10.1111/j.1742-4658.2007.05720.x. Epub 2007 Mar 9.
Quantitation of superoxide radical (O (2)(-).) production at the site of radical generation remains challenging. A simple method to detect nanomolar to micromolar levels of superoxide radical in aqueous solution has been developed and optimized. This method is based on the efficient trapping of O(2)(-). using a novel fluorescent probe (2-chloro-1,3-dibenzothiazolinecyclohexene), coupled with a spectra character-signaling increase event. A high-specificity and high-sensitivity fluorescent probe was synthesized in-house and used to image O(2)(-). in living cells. Better selectivity for O(2)(-). over competing cellular reactive oxygen species and some biological compounds illustrates the advantages of our method. Under optimal conditions, the linear calibration range for superoxide anion radicals was 5.03 x 10(-9)-3.33 x 10(-6) M. The detection limit was 1.68 x 10(-9) M. Fluorescence images of probe-stained macrophages stimulated with 4beta-phorbol 12-myristate 13-acetate were obtained successfully using a confocal laser scanning microscope.
对超氧阴离子自由基（O₂⁻）产生部位进行定量分析仍然具有挑战性。一种用于检测水溶液中纳摩尔至微摩尔水平超氧阴离子自由基的简单方法已得到开发和优化。该方法基于使用新型荧光探针（2-氯-1,3-二苯并噻唑啉环己烯）对O₂⁻的有效捕获，以及光谱特征信号增加事件。我们自行合成了一种高特异性和高灵敏度的荧光探针，并用于对活细胞中的O₂⁻进行成像。与竞争性细胞活性氧物种和一些生物化合物相比，该方法对O₂⁻具有更好的选择性，这说明了我们方法的优势。在最佳条件下，超氧阴离子自由基的线性校准范围为5.03×10⁻⁹ - 3.33×10⁻⁶ M。检测限为1.68×10⁻⁹ M。使用共聚焦激光扫描显微镜成功获得了用4β-佛波醇12-肉豆蔻酸酯13-乙酸酯刺激的探针染色巨噬细胞的荧光图像。
