利用新型红移荧光蛋白进行同步双色荧光寿命成像。
Simultaneous dual-color fluorescence lifetime imaging with novel red-shifted fluorescent proteins.
作者信息
Laviv Tal, Kim Benjamin B, Chu Jun, Lam Amy J, Lin Michael Z, Yasuda Ryohei
机构信息
Neuronal Signal Transduction Group, Max Planck Florida Institute for Neuroscience, Jupiter, Florida, USA.
Department of Bioengineering, Stanford University, Stanford, California, USA.
出版信息
Nat Methods. 2016 Dec;13(12):989-992. doi: 10.1038/nmeth.4046. Epub 2016 Oct 31.
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.
摘要
我们描述了一种为双色荧光寿命成像（FLIM）优化的红移荧光共振能量转移（FRET）对。该对使用了一种新开发的FRET供体，单体青色可激发红色荧光蛋白（mCyRFP1），它具有大的斯托克斯位移和单指数荧光寿命衰减。当与基于增强型绿色荧光蛋白（EGFP）的生物传感器一起使用时，这一新的对能够在经历结构可塑性的单个树突棘中同时成像两种信号分子的活性。
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