荧光
生物分子
显微镜
量子产额
生物物理学
化学
荧光显微镜
超分辨显微术
亮度
光漂白
光化学
材料科学
纳米技术
分子
生物
光学
有机化学
物理
作者
Jonathan B. Grimm,Brian P. English,Jiji Chen,Joel P Slaughter,Zhengjian Zhang,Andrey Revyakin,Ronak Patel,John J. Macklin,Davide Normanno,Robert H. Singer,Timothée Lionnet,Luke D. Lavis
出处
期刊:Nature Methods
[Springer Nature]
日期:2015-01-19
卷期号:12 (3): 244-250
被引量:1241
摘要
A simple and general chemical structure change to a panel of cell-permeable small-molecule fluorophores increases their brightness and photostability, which will enable improved single-molecule studies and super-resolution imaging. Specific labeling of biomolecules with bright fluorophores is the keystone of fluorescence microscopy. Genetically encoded self-labeling tag proteins can be coupled to synthetic dyes inside living cells, resulting in brighter reporters than fluorescent proteins. Intracellular labeling using these techniques requires cell-permeable fluorescent ligands, however, limiting utility to a small number of classic fluorophores. Here we describe a simple structural modification that improves the brightness and photostability of dyes while preserving spectral properties and cell permeability. Inspired by molecular modeling, we replaced the N,N-dimethylamino substituents in tetramethylrhodamine with four-membered azetidine rings. This addition of two carbon atoms doubles the quantum efficiency and improves the photon yield of the dye in applications ranging from in vitro single-molecule measurements to super-resolution imaging. The novel substitution is generalizable, yielding a palette of chemical dyes with improved quantum efficiencies that spans the UV and visible range.
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