6533b7d8fe1ef96bd126b5d6
RESEARCH PRODUCT
Bioorthogonal red and far-red fluorogenic probes for wash-free live-cell and super-resolution microscopy
Miao YuFelix BraunWeijie ChiEdward A. LemkeTiago BuckupKristin S. GrussmayerZhibin ZhangZhibin ZhangAntoni J. GralakRichard WombacherRichard WombacherDirk-peter HertenDirk-peter HertenDirk-peter HertenKlaus YserentantChristoph MayerMichael J. ZieglerMichael J. ZieglerFrank RomingerMarvin BuschPhilipp WertherVytautas NavikasAleksandra RadenovicXiaogang Liusubject
chemistry.chemical_compoundFluorescence-lifetime imaging microscopyTetrazineFluorophoreQuenching (fluorescence)chemistrySuper-resolution microscopySTED microscopyContext (language use)Bioorthogonal chemistryCombinatorial chemistrydescription
AbstractSmall-molecule fluorophores enable the observation of biomolecules in their native context with fluorescence microscopy. Specific labelling via bioorthogonal tetrazine chemistry confers minimal label size and rapid labelling kinetics. At the same time, fluorogenic tetrazine-dye conjugates exhibit efficient quenching of dyes prior to target binding. However, live-cell compatible long-wavelength fluorophores with strong fluorogenicity have been difficult to realize. Here, we report close proximity tetrazine-dye conjugates with minimal distance between tetrazine and fluorophore. Two synthetic routes give access to a series of cell permeable and impermeable dyes including highly fluorogenic far-red emitting derivatives with electron exchange as dominant excited state quenching mechanism. We demonstrate their potential for live-cell imaging in combination with unnatural amino acids, wash-free multi-colour and super-resolution STED and SOFI imaging. These dyes pave the way for advanced fluorescence imaging of biomolecules with minimal label size.
| year | journal | country | edition | language |
|---|---|---|---|---|
| 2020-08-07 |