刃天青与试卤灵酸度响应性的光谱研究

A Spectroscopic Investigation of pH Sensitivities of Resorufin and Resazurin

摘要:

刃天青是单分子单粒子荧光显微技术中一种常用的荧光探针。然而其荧光性质随环境的变化会发生明显的变化，这导致单分子荧光信号的不稳定性。本文基于pH对荧光探针发光性质的影响，研究了缓冲溶液pH与浓度对荧光强度的影响，获得了荧光最优化的条件。进一步构建了数学模型，结合理论计算，获取了其荧光强度与氢离子的依赖关系，并对pH的响应性能给出了理论解释。结果表明，随着pH的增加，荧光强度先增大后减小。理论研究表明，荧光猝灭与刃天青和间苯二酚的质子化有关。

Abstract:

Resazurin is a commonly used dye for single molecule fluorescence microscopy. However， its fluorescence properties change obviously with the change of environment， which leads to the instability of single molecule fluorescence signal. Herein， based on the influence of pH on luminescence properties of fluorescent probe， the fluorescence properties of resorufin and resazurin in phosphate buffer solution were reported and the fluorescence optimization conditions were obtained. In addition， a mathematical model was further built and the dependence between fluorescence intensity and hydrogen ion was obtained combining with theoretical calculation， which provided a theoretical explanation for pH response performance. The results showed that based on the increase of pH， the fluorescence intensity increased first and then decreased. Theoretical studies showed that the fluorescence quenching correlates with the protonation of the resazurin and resorufin.