title: Simultaneous dual-color fluorescence lifetime imaging with novel red-shifted fluorescent proteins.
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.
authors: Neuronal Signal Transduction Group, Max Planck Florida Institute for Neuroscience, Jupiter, Florida, USA.
authors: Tal
authors: T
authors: Laviv
authors: Department of Bioengineering, Stanford University, Stanford, California, USA.
authors: Benjamin B
authors: BB
authors: Kim
authors: Department of Bioengineering, Stanford University, Stanford, California, USA.|Department of Pediatrics, Stanford University, Stanford, California, USA.
authors: Jun
authors: J
authors: Chu
authors: Department of Bioengineering, Stanford University, Stanford, California, USA.|Department of Pediatrics, Stanford University, Stanford, California, USA.
authors: Amy J
authors: AJ
authors: Lam
authors: Department of Bioengineering, Stanford University, Stanford, California, USA.|Department of Pediatrics, Stanford University, Stanford, California, USA.|Department of Neurobiology, Stanford University, Stanford, California, USA.
authors: Michael Z
authors: MZ
authors: Lin
authors: Neuronal Signal Transduction Group, Max Planck Florida Institute for Neuroscience, Jupiter, Florida, USA.
authors: Ryohei
authors: R
authors: Yasuda
mesh_terms: D000818:Animals; D015374:Biosensing Techniques / Q000379:methods*; D018274:Electroporation; D004721:Endoplasmic Reticulum / Q000378:metabolism; D005260:Female; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods*; D005456:Fluorescent Dyes / Q000737:chemistry* / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000737:chemistry* / Q000378:metabolism; D057809:HEK293 Cells; D006367:HeLa Cells; D006801:Humans; D008164:Luminescent Proteins / Q000737:chemistry* / Q000378:metabolism; D051379:Mice; D036641:Microscopy, Fluorescence, Multiphoton; D061848:Optical Imaging / Q000379:methods*; D038761:Photobleaching; D011247:Pregnancy; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000378:metabolism; D014162:Transfection; D000097573:Red Fluorescent Protein
chemical_list: D005456:Fluorescent Dyes; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins; D011993:Recombinant Fusion Proteins; D000097573:Red Fluorescent Protein