Open AccessChemical reactivation of resin-embedded pHuji adds red for simultaneous two-color imaging with EGFP
Author(s) -
Wenyan Guo,
Xiuli Liu,
Yurong Liu,
Yadong Gang,
Xiaobin He,
Jia Yao,
FangFang Yin,
Pei Li,
Fei Huang,
Haiyan Zhou,
Xiaojun Wang,
Hui Gong,
Qingming Luo,
Fuqiang Xu,
Shaoqun Zeng
Publication year - 2017
Publication title -
biomedical optics express
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.362
H-Index - 86
ISSN - 2156-7085
DOI - 10.1364/boe.8.003281
Subject(s) - fluorescence , green fluorescent protein , fluorescent protein , biophysics , fluorescence lifetime imaging microscopy , chemistry , materials science , optics , biochemistry , biology , physics , gene
The pH-sensitive fluorescent proteins enabling chemical reactivation in resin are useful tools for fluorescence microimaging. EGFP or EYFP is good for such applications. For simultaneous two-color imaging, a suitable red fluorescent protein is an urgent need. Here a pH-sensitive red fluorescent protein, pHuji, is selected and verified to remain pH-sensitive in HM20 resin. We observe 183% fluorescence intensity of pHuji in resin-embeded mouse brain and 29.08-fold fluorescence intensity of reactivated pHuji compared to the quenched state. pHuji and EGFP can be quenched and chemically reactivated simultaneously in resin, thus enabling simultaneous two-color micro-optical sectioning tomography of resin-embedded mouse brain. This method may greatly facilitate the visualization of neuronal morphology and neural circuits to promote understanding of the structure and function of the brain.