ArticleEuropean biophysics journal : EBJ2008
satFRET: estimation of Förster resonance energy transfer by acceptor saturation.
Article in European biophysics journal : EBJ, 2008. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 11 papers.
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Who cites it
11 citing papers in PubMed, 50 citations in OpenAlex.
- Molecular Interaction of the Coumarin Derivative Umbelliferone with Cu/Zn-SOD1 and DNA: Insights into Binding Mode and Antioxidant Mechanism.Journal of fluorescence · 2025Article
- Dimerization of melanocortin 4 receptor controls puberty onset and body size polymorphism.Frontiers in endocrinology · 2023Article
- Dual-color metal-induced and Förster resonance energy transfer for cell nanoscopy.Molecular biology of the cell · 2018Article
- Combining Graphical and Analytical Methods with Molecular Simulations To Analyze Time-Resolved FRET Measurements of Labeled Macromolecules Accurately.The journal of physical chemistry. B · 2017Article
- Steady-state acceptor fluorescence anisotropy imaging under evanescent excitation for visualisation of FRET at the plasma membrane.PloS one · 2014Article
- Single cell optical imaging and spectroscopy.Chemical reviews · 2013Review
- OS-FRET: a new one-sample method for improved FRET measurements.Biochemistry · 2011Article
- Article
- The design of Förster (fluorescence) resonance energy transfer (FRET)-based molecular sensors for Ran GTPase.Methods (San Diego, Calif.) · 2010Review
- FRET-enabled optical modulation for high sensitivity fluorescence imaging.Journal of the American Chemical Society · 2010Article
- The potential of optical proteomic technologies to individualize prognosis and guide rational treatment for cancer patients.Targeted oncology · 2009Review
Corrections and comments
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Authors and funding
7 authors at 4 institutions in 3 countries.
Funding
Abstract
We demonstrate theoretically and experimentally the quantification of Förster resonance energy transfer (FRET) by direct and systematic saturation of the excited state of acceptor molecules. This version of acceptor depletion methods for FRET estimation, denoted as "satFRET" is reversible and suitable for time-resolved measurements. The technique was investigated theoretically using the steady-state solution of the differential equation system of donor and acceptor molecular states. The influence of acceptor photobleaching during measurement was included in the model. Experimental verification was achieved with the FRET-pair Alexa 546- Alexa 633 loaded on particles in different stoichiometries and measured in a confocal microscope. Estimates of energy transfer efficiency by excited state saturation were compared to those obtained by measurements of sensitised emission and acceptor photobleaching. The results lead to a protocol that allows time-resolved FRET measurements of fixed and living cells on a conventional confocal microscope. This procedure was applied to fixed Chinese hamster ovary cells containing a cyan fluorescent protein and yellow fluorescent protein pair. The time resolution of the technique was demonstrated in a live T cell activation assay comparing the FRET efficiencies measured using a genetically encoded green and red fluorescent protein biosensor for GTP/GDP turnover to those measured by acceptor photobleaching of fixed cells.
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