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


Journal Article

© 2017 Optical Society of America. 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.

Full Text

Duke Authors

Cited Authors

  • Guo, W; Liu, X; Liu, Y; Gang, Y; He, X; Jia, Y; Yin, F; Li, P; Huang, F; Zhou, H; Wang, X; Gong, H; Luo, Q; Xu, F; Zeng, S

Published Date

  • July 1, 2017

Published In

Volume / Issue

  • 8 / 7

Start / End Page

  • 3281 - 3288

Electronic International Standard Serial Number (EISSN)

  • 2156-7085

Digital Object Identifier (DOI)

  • 10.1364/BOE.8.003281

Citation Source

  • Scopus