一种基于 FRET 的比率型化学传感器，用于体外细胞 pH 值的荧光分析。
An FRET-based ratiometric chemosensor for in vitro cellular fluorescence analyses of pH.
机构信息
Center for Biosignatures Discovery Automation, Biodesign Institute, Arizona State University, Tempe, AZ 85287, USA.
出版信息
Biomaterials. 2012 Jan;33(1):171-80. doi: 10.1016/j.biomaterials.2011.09.053. Epub 2011 Oct 7.
Abstract
Ratiometric fluorescence sensing is an important technique for precise and quantitative analysis of biological events occurring under complex conditions by simultaneously recording fluorescence intensities at two wavelengths and calculating their ratios. Herein, we design a ratiometric chemosensor for pH that is based on photo-induced electron transfer (PET) and binding-induced modulation of fluorescence resonance energy transfer (FRET) mechanisms. This ratiometric chemosensor was constructed by introduction of a pH-insensitive coumarin fluorophore as an FRET donor into a pH-sensitive amino-naphthalimide derivative as the FRET acceptor. The sensor exhibited clear dual-mission signal changes in blue and green spectral windows upon pH changes. The pH sensor was applied for not only measuring cellular pH, but also for visualizing stimulus-responsive changes of intracellular pH values.
摘要
比率荧光传感是一种重要的技术，可通过同时记录两个波长的荧光强度并计算它们的比值，对复杂条件下发生的生物事件进行精确和定量分析。在此，我们设计了一种基于光诱导电子转移（PET）和结合诱导的荧光共振能量转移（FRET）机制调制的 pH 比率型化学传感器。该比率型化学传感器通过将 pH 不敏感的香豆素荧光团作为 FRET 供体引入 pH 敏感的氨基-萘酰亚胺衍生物作为 FRET 受体来构建。该传感器在 pH 变化时在蓝色和绿色光谱窗口中表现出明显的双发射信号变化。该 pH 传感器不仅可用于测量细胞 pH，还可用于可视化细胞内 pH 值的刺激响应变化。
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