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

Laviv, Tal; Kim, Benjamin B; Chu, Jun; Lam, Amy J; Lin, Michael Z; Yasuda, Ryohei · Nat Methods · 2016

basic_science · Level V

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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.

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