Imaging the Redox States of Live Cells with the Time-Resolved Fluorescence of Genetically Encoded Biosensors
Imaging the Redox States of Live Cells with the Time-Resolved Fluorescence of Genetically Encoded Biosensors
复制标题
利用基因编码生物传感器的时间分辨荧光对活细胞的氧化还原状态进行成像
DOI：
10.1021/acs.analchem.8b04292
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发表时间：
2019
影响因子：
7.4
通讯作者：
Tian Yang
中科院分区：
文献类型：
--
作者：
Li Lei;Zhang Changcheng;Wang Peng;Wang Aoxue;Zhou Jiasheng;Chen Guoqing;Xu Jianhua;Yang Yi;Zhao Yuzheng;Zhang Sanjun;Tian Yang
细胞中的氧化还原环境影响许多重要的生理和病理过程。在这项研究中，时间分辨荧光的巯基氧化还原敏感传感器的脊椎动物荧光蛋白UnaG，roUnaG，进行了研究，沿着的应用程序的时间分辨荧光的roUnaG成像的线粒体，细胞质和细胞核的氧化还原状态在活细胞。roUnaG的时间分辨荧光图像清楚地表明，有效的抗癌化合物KP 372 -1诱导极端氧化应激。在活化的巨噬细胞中观察到的更应激的氧化还原状态进一步证明了具有时间分辨荧光的roUnaG的有效性。为了进行比较，还捕获了其他四种常用氧化还原生物传感器（roGFP 1、HyPer、HyPerRed和rxRFP）的时间分辨荧光图像。时间分辨荧光允许一个固有的比率测量的生物传感器与一个激发波长，并提供了新的机会，生物成像。
Redox environments in cells influence many important physiological and pathological processes. In this study, the time-resolved fluorescence of a recently reported thiol redox-sensitive sensor based on vertebrate fluorescent protein UnaG, roUnaG, was studied, along with the application of the time-resolved fluorescence of roUnaG to image the redox states of the mitochondria, cytoplasm, and nucleus in live cells. Time-resolved fluorescence images of roUnaG clearly demonstrated that potent anticancer compound KP372-1 induced extreme oxidative stress. A more stressful redox state observed in activated macrophages further demonstrated the validity of roUnaG with time-resolved fluorescence. For comparison, time-resolved fluorescence images of four other frequently used redox biosensors (roGFP1, HyPer, HyPerRed, and rxRFP) were also captured. The time-resolved fluorescence allows an intrinsically ratiometric measurement for biosensors with one excitation wavelength and provides new opportunities for bioimaging.