Unknown

Dataset Information

0

A general method to improve fluorophores for live-cell and single-molecule microscopy.


ABSTRACT: 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.

SUBMITTER: Grimm JB 

PROVIDER: S-EPMC4344395 | biostudies-literature | 2015 Mar

REPOSITORIES: biostudies-literature

altmetric image

Publications


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 brightnes  ...[more]

Similar Datasets

| S-EPMC6476321 | biostudies-literature
| S-EPMC8154212 | biostudies-literature
| S-EPMC5621985 | biostudies-literature
| S-EPMC5299821 | biostudies-literature
| S-EPMC7582769 | biostudies-literature
| S-EPMC4142238 | biostudies-literature
| S-EPMC6420572 | biostudies-literature
| S-EPMC7714787 | biostudies-literature
| S-EPMC3873762 | biostudies-literature
| S-EPMC3150364 | biostudies-literature