Multiplexed 3D FRET imaging in deep tissue of live embryos.

Published

Journal Article

Current deep tissue microscopy techniques are mostly restricted to intensity mapping of fluorophores, which significantly limit their applications in investigating biochemical processes in vivo. We present a deep tissue multiplexed functional imaging method that probes multiple Förster resonant energy transfer (FRET) sensors in live embryos with high spatial resolution. The method simultaneously images fluorescence lifetimes in 3D with multiple excitation lasers. Through quantitative analysis of triple-channel intensity and lifetime images, we demonstrated that Ca(2+) and cAMP levels of live embryos expressing dual FRET sensors can be monitored simultaneously at microscopic resolution. The method is compatible with a broad range of FRET sensors currently available for probing various cellular biochemical functions. It opens the door to imaging complex cellular circuitries in whole live organisms.

Full Text

Cited Authors

  • Zhao, M; Wan, X; Li, Y; Zhou, W; Peng, L

Published Date

  • September 21, 2015

Published In

Volume / Issue

  • 5 /

Start / End Page

  • 13991 -

PubMed ID

  • 26387920

Pubmed Central ID

  • 26387920

Electronic International Standard Serial Number (EISSN)

  • 2045-2322

International Standard Serial Number (ISSN)

  • 2045-2322

Digital Object Identifier (DOI)

  • 10.1038/srep13991

Language

  • eng