A general method to improve fluorophores for live-cell and single-molecule microscopy.
A general method to improve fluorophores for live-cell and single-molecule microscopy.
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DOI：
10.1038/nmeth.3256
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发表时间：
2015-03
期刊：
影响因子：
48
通讯作者：
Lavis, Luke D.
中科院分区：
文献类型：
--
作者：
Grimm, Jonathan B.;English, Brian P.;Chen, Jiji;Slaughter, Joel P.;Zhang, Zhengjian;Revyakin, Andrey;Patel, Ronak;Macklin, John J.;Normanno, Davide;Singer, Robert H.;Lionnet, Timothee;Lavis, Luke D.
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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影响因子：
48
作者：
Dempsey, Graham T.;Vaughan, Joshua C.;Chen, Kok Hao;Bates, Mark;Zhuang, Xiaowei
通讯作者：
Zhuang, Xiaowei
影响因子：
48
作者：
Abrahamsson, Sara;Chen, Jiji;Hajj, Bassam;Stallinga, Sjoerd;Katsov, Alexander Y.;Wisniewski, Jan;Mizuguchi, Gaku;Soule, Pierre;Mueller, Florian;Darzacq, Claire Dugast;Darzacq, Xavier;Wu, Carl;Bargmann, Cornelia I.;Agard, David A.;Dahan, Maxime;Gustafsson, Mats G. L.
通讯作者：
Gustafsson, Mats G. L.
影响因子：
3.4
作者：
Bosch, Peter J.;Correa, Ivan R., Jr.;Subramaniam, Vinod
通讯作者：
Subramaniam, Vinod
影响因子：
3.8
作者：
KUBOTA, Y;STEINER, RF
通讯作者：
STEINER, RF
影响因子：
14.7
作者：
Ha T;Tinnefeld P
通讯作者：
Tinnefeld P