Chemical reactivation of resin-embedded pHuji adds red for simultaneous two-color imaging with EGFP

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 prot...

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Bibliographic Details
Published in:Biomedical optics express 2017-07, Vol.8 (7), p.3281-3288
Main Authors: Guo, Wenyan, Liu, Xiuli, Liu, Yurong, Gang, Yadong, He, Xiaobin, Jia, Yao, Yin, Fangfang, Li, Pei, Huang, Fei, Zhou, Hongfu, Wang, Xiaojun, Gong, Hui, Luo, Qingming, Xu, Fuqiang, Zeng, Shaoqun
Format: Article
Language:English
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Summary: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.
ISSN:2156-7085
2156-7085
DOI:10.1364/BOE.8.003281