Structure-guided evolution of cyan fluorescent proteins towards a quantum yield of 93%
- J. Goedhart, D. von Stetten, A. Royant
- BiologyNature Communications
- 1 January 2012
A new CFP, mTurquoise2, is developed, which displays a high-fluorescence quantum yield and a long mono-exponential fluorescence lifetime and is the preferable cyan variant of green fluorescent protein for long-term imaging and as donor for Förster resonance energy transfer to a yellow fluorescent protein.
Fluorescence lifetime imaging by time‐correlated single‐photon counting
- W. Becker, A. Bergmann, M. Hink, K. König, K. Benndorf, C. Biskup
- PhysicsMicroscopy research and technique (Print)
We present a time‐correlated single photon counting (TCPSC) technique that allows time‐resolved multi‐wavelength imaging in conjunction with a laser scanning microscope and a pulsed excitation…
mScarlet: a bright monomeric red fluorescent protein for cellular imaging
- D. Bindels, Lindsay Haarbosch, T. Gadella
- ChemistryNature Methods
- 21 November 2016
The engineering of mScarlet is reported, a truly monomeric red fluorescent protein with record brightness, quantum yield, and fluorescence lifetime and it is especially useful as a Förster resonance energy transfer (FRET) acceptor in ratiometric imaging.
A mTurquoise-Based cAMP Sensor for Both FLIM and Ratiometric Read-Out Has Improved Dynamic Range
- J. Klarenbeek, J. Goedhart, M. Hink, T. Gadella, K. Jalink
- ChemistryPLoS ONE
- 29 April 2011
It is shown that TEpacVV appears optimal for detection both by FLIM and SE, that it has outstanding FRET span and signal-to-noise ratio, and improved photostability, and should become the cAMP sensor of choice for new experiments, both forFLIM and ratiometric detection.
Spatial regulation of Fus3 MAP kinase activity through a reaction-diffusion mechanism in yeast pheromone signalling
- Celine I. Maeder, M. Hink, A. Kinkhabwala, Reinhard Mayr, P. Bastiaens, M. Knop
- BiologyNature Cell Biology
- 1 November 2007
Investigation of the distribution of active Fus3 (Fus3PP) across the cytoplasm revealed a gradient of FUS3PP activity emanating from the tip of the mating projection, suggesting that Propagation of signalling from the shmoo is, therefore, spatially constrained by a gradient-generating reaction-diffusion mechanism.
Bright cyan fluorescent protein variants identified by fluorescence lifetime screening
- J. Goedhart, L. V. Weeren, M. Hink, N. Vischer, K. Jalink, T. Gadella
- Biology, PhysicsNature Methods
- 1 February 2010
A strategy to screen excited state lifetimes identified cyan fluorescent proteins with long fluorescence lifetimes and high quantum yields, making one variant, mTurquoise, an excellent fluorescence resonance energy transfer (FRET) donor.
Structural Dynamics of Green Fluorescent Protein Alone and Fused with a Single Chain Fv Protein*
The dynamic properties of GFP alone and fused to a single chain antibody raised against lipopolysaccharide of the outer cell wall of Gram-negative bacteria were investigated and the scFv moiety was functional as was proven in binding assays.
An orange fluorescent protein with a large Stokes shift for single-excitation multicolor FCCS and FRET imaging.
- D. Shcherbakova, M. Hink, L. Joosen, T. Gadella, V. Verkhusha
- PhysicsJournal of the American Chemical Society
- 9 May 2012
The LSSmOrange mutagenesis, low-temperature, and isotope effect studies revealed a proton relay for the excited-state proton transfer responsible for the LSS phenotype and developed four-color single-laser fluorescence cross-correlation spectroscopy, solely based on FPs.
Fluorescence correlation microscopy of cells in the presence of autofluorescence.
Effects of Refractive Index and Viscosity on Fluorescence and Anisotropy Decays of Enhanced Cyan and Yellow Fluorescent Proteins
Time-resolved fluorescence measurements of the enhanced forms of ECFP and EYFP in water–glycerol mixtures were performed to quantify the effects of the refractive index and viscosity on the fluorescence lifetimes of these proteins.