ArticleBiophysical journal2000
Sensitivity enhancement in fluorescence correlation spectroscopy of multiple species using time-gated detection.
Article in Biophysical journal, 2000. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 23 papers.
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Who cites it
23 citing papers in PubMed.
- Breaking the Low Concentration Barrier of Single-Molecule Fluorescence Quantification to the Sub-Picomolar Range.Small methods · 2026Article
- Correction of Background in Fluorescence Correlation Spectroscopy for Accurate Determination of Particle Number.Biomolecules · 2025Article
- Shrinking gate fluorescence correlation spectroscopy yields equilibrium constants and separates photophysics from structural dynamics.Proceedings of the National Academy of Sciences of the United States of America · 2023Article
- Pulsed interleaved excitation-based line-scanning spatial correlation spectroscopy (PIE-lsSCS).Scientific reports · 2018Article
- STED-FLCS: An Advanced Tool to Reveal Spatiotemporal Heterogeneity of Molecular Membrane Dynamics.Nano letters · 2015Article
- Live-cell observation of cytosolic HIV-1 assembly onset reveals RNA-interacting Gag oligomers.The Journal of cell biology · 2015Article
- Pulsed interleaved excitation fluctuation imaging.Biophysical journal · 2013Article
- Fluorescence Lifetime Correlation Spectroscopy (FLCS): concepts, applications and outlook.International journal of molecular sciences · 2012Review
- Factors affecting the quantification of biomolecular interactions by fluorescence cross-correlation spectroscopy.Biophysical journal · 2012Article
- Filtered FCS: species auto- and cross-correlation functions highlight binding and dynamics in biomolecules.Chemphyschem : a European journal of chemical physics and physical chemistry · 2012Article
- Early amyloidogenic oligomerization studied through fluorescence lifetime correlation spectroscopy.International journal of molecular sciences · 2012Article
- Characterization of protein adsorption onto FePt nanoparticles using dual-focus fluorescence correlation spectroscopy.Beilstein journal of nanotechnology · 2011Article
- Detecting intramolecular dynamics and multiple Förster resonance energy transfer states by fluorescence correlation spectroscopy.The journal of physical chemistry. B · 2010Article
- Quantitative analysis of the protein corona on FePt nanoparticles formed by transferrin binding.Journal of the Royal Society, Interface · 2010Article
- Transient anomalous subdiffusion: effects of specific and nonspecific probe binding with actin gels.The journal of physical chemistry. B · 2010Article
- On the resolution capabilities and limits of fluorescence lifetime correlation spectroscopy (FLCS) measurements.Journal of fluorescence · 2010Article
- A quantitative fluorescence study of protein monolayer formation on colloidal nanoparticles.Nature nanotechnology · 2009Article
- spFRET using alternating excitation and FCS reveals progressive DNA unwrapping in nucleosomes.Biophysical journal · 2009Article
- Fluorescence lifetime correlation spectroscopy.Journal of fluorescence · 2007Article
- Pulsed interleaved excitation.Biophysical journal · 2005Article
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5 authors.
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No grant is acknowledged in the PubMed record.
Abstract
Fluorescence correlation spectroscopy (FCS) is a powerful technique to measure chemical reaction rates and diffusion coefficients of molecules in thermal equilibrium. The capabilities of FCS can be enhanced by measuring the energy, polarization, or delay time between absorption and emission of the collected fluorescence photons in addition to their arrival times. This information can be used to change the relative intensities of multiple fluorescent species in FCS measurements and, thus, the amplitude of the intensity autocorrelation function. Here we demonstrate this strategy using lifetime gating in FCS experiments. Using pulsed laser excitation and laser-synchronized gating in the detection channel, we suppress photons emitted within a certain time interval after excitation. Three applications of the gating technique are presented: suppression of background fluorescence, simplification of FCS reaction studies, and investigation of lifetime heterogeneity of fluorescently labeled biomolecules. The usefulness of this technique for measuring forward and backward rates of protein fluctuations in equilibrium and for distinguishing between static and dynamic heterogeneity makes it a promising tool in the investigation of chemical reactions and conformational fluctuations in biomolecules.
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