A general method to improve fluorophores for live-cell and single-molecule microscopy
Abstrak
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.
Penulis (12)
J. Grimm
Brian P. English
Jiji Chen
Joel P Slaughter
Zhengjian Zhang
A. Revyakin
Ronak Patel
J. Macklin
Davide Normanno
R. Singer
T. Lionnet
L. Lavis
Akses Cepat
- Tahun Terbit
- 2014
- Bahasa
- en
- Total Sitasi
- 1271×
- Sumber Database
- Semantic Scholar
- DOI
- 10.1038/nmeth.3256
- Akses
- Open Access ✓