Cy3-Cy5 Covalent Heterodimers for Single-Molecule Photoswitching
- 28 August 2008
- journal article
- letter
- Published by American Chemical Society (ACS) in The Journal of Physical Chemistry B
- Vol. 112 (38) , 11878-11880
- https://doi.org/10.1021/jp806698p
Abstract
Covalent heterodimers of the Cy3 and Cy5 fluorophores have been prepared from commercially available starting materials and characterized at the single-molecule level. This system behaves as a discrete molecular photoswitch, in which photoexcitation of the Cy5 results in fluorescence emission or, with a much lower probability, causes the Cy5 to enter into a long-lived, but metastable, dark state. Photoinduced recovery of the emissive Cy5 is achieved by very low intensity excitation (5 W cm−2) of the Cy3 fluorophore at a shorter wavelength. A similar system consisting of proximal, but not covalently linked, Cy3 and Cy5 has found application in stochastic optical reconstruction microscopy (STORM), a single-molecule localization-based technique for super-resolution imaging that requires photoswitching. The covalent Cy3-Cy5 heterodimers described herein eliminate the need for probabilistic methods of situating the Cy3 and Cy5 in close proximity to enable photoswitching. As proof of principle, these heterodimers have been applied to super-resolution imaging of the tubular stalk structures of live Caulobacter crescentus bacterial cells.Keywords
This publication has 11 references indexed in Scilit:
- HaloTag: A Novel Protein Labeling Technology for Cell Imaging and Protein AnalysisACS Chemical Biology, 2008
- Three-Dimensional Super-Resolution Imaging by Stochastic Optical Reconstruction MicroscopyScience, 2008
- Nonblinking and long-lasting single-molecule fluorescence imagingNature Methods, 2006
- Sub-diffraction-limit imaging by stochastic optical reconstruction microscopy (STORM)Nature Methods, 2006
- Novel synthesis of [18F]-fluorobenzene and pyridinecarbohydrazide-folates as potential PET radiopharmaceuticalsJournal of Labelled Compounds and Radiopharmaceuticals, 2005
- A Reagentless Biosensing Assembly Based on Quantum Dot–Donor Förster Resonance Energy TransferAdvanced Materials, 2005
- Short-Range Spectroscopic Ruler Based on a Single-Molecule Optical SwitchPhysical Review Letters, 2005
- Carbocyanine Dyes as Efficient Reversible Single-Molecule Optical SwitchJournal of the American Chemical Society, 2005
- A general method for the covalent labeling of fusion proteins with small molecules in vivoNature Biotechnology, 2002
- Autofluorescent Proteins in Single-Molecule Research: Applications to Live Cell Imaging MicroscopyBiophysical Journal, 2001