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Nat Methods. 2016 Dec;13(12):989-992. doi: 10.1038/nmeth.4046. Epub 2016 Oct 31.

Simultaneous dual-color fluorescence lifetime imaging with novel red-shifted fluorescent proteins.

Author information

1
Neuronal Signal Transduction Group, Max Planck Florida Institute for Neuroscience, Jupiter, Florida, USA.
2
Department of Bioengineering, Stanford University, Stanford, California, USA.
3
Department of Pediatrics, Stanford University, Stanford, California, USA.
4
Department of Neurobiology, Stanford University, Stanford, California, USA.

Abstract

We describe a red-shifted fluorescence resonance energy transfer (FRET) pair optimized for dual-color fluorescence lifetime imaging (FLIM). This pair utilizes a newly developed FRET donor, monomeric cyan-excitable red fluorescent protein (mCyRFP1), which has a large Stokes shift and a monoexponential fluorescence lifetime decay. When used together with EGFP-based biosensors, the new pair enables simultaneous imaging of the activities of two signaling molecules in single dendritic spines undergoing structural plasticity.

PMID:
27798609
PMCID:
PMC5322478
DOI:
10.1038/nmeth.4046
[Indexed for MEDLINE]
Free PMC Article

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