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A tunable fluorescent timer method for imaging spatial‐temporal protein dynamics using light‐driven photoconvertible protein
Author(s) -
Zhu Xinxin,
Zhang Luyuan,
Kao YaTing,
Xu Fang,
Min Wei
Publication year - 2015
Publication title -
journal of biophotonics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.877
H-Index - 66
eISSN - 1864-0648
pISSN - 1864-063X
DOI - 10.1002/jbio.201300174
Subject(s) - timer , fluorescent protein , fluorescence , biological system , dynamics (music) , temporal resolution , biophysics , fluorescent light , computer science , nanotechnology , green fluorescent protein , chemistry , materials science , physics , optics , biology , computer hardware , acoustics , biochemistry , gene , microcontroller
Cellular function is largely determined by protein behaviors occurring in both space and time. While regular fluorescent proteins can only report spatial locations of the target inside cells, fluorescent timers have emerged as an invaluable tool for revealing coupled spatial‐temporal protein dynamics. Existing fluorescent timers are all based on chemical maturation. Herein we propose a light‐driven timer concept that could report relative protein ages at specific sub‐cellular locations, by weakly but chronically illuminating photoconvertible fluorescent proteins inside cells. This new method exploits light, instead of oxygen, as the driving force. Therefore its timing speed is optically tunable by adjusting the photoconverting laser intensity. We characterized this light‐driven timer method both in vitro and in vivo and applied it to image spatiotemporal distributions of several proteins with different lifetimes. This novel timer method thus offers a flexible “ruler” for studying temporal hierarchy of spatially ordered processes with exquisite spatial‐temporal resolution. (© 2015 WILEY‐VCH Verlag GmbH &Co. KGaA, Weinheim)