TY - JOUR
T1 - Imaging the redox states of live cells with the time-resolved fluorescence of genetically encoded biosensors
AU - Li, Lei
AU - Zhang, Changcheng
AU - Wang, Peng
AU - Wang, Aoxue
AU - Zhou, Jiasheng
AU - Chen, Guoqing
AU - Xu, Jianhua
AU - Yang, Yi
AU - Zhao, Yuzheng
AU - Zhang, Sanjun
AU - Tian, Yang
N1 - Publisher Copyright:
© 2019 American Chemical Society.
PY - 2019/3/19
Y1 - 2019/3/19
N2 - 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.
AB - 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.
UR - https://www.scopus.com/pages/publications/85063152596
U2 - 10.1021/acs.analchem.8b04292
DO - 10.1021/acs.analchem.8b04292
M3 - 文章
C2 - 30777423
AN - SCOPUS:85063152596
SN - 0003-2700
VL - 91
SP - 3869
EP - 3876
JO - Analytical Chemistry
JF - Analytical Chemistry
IS - 6
ER -