Fluorescence correlation spectroscopy: the technique and its applications

  title={Fluorescence correlation spectroscopy: the technique and its applications},
  author={Oleg Krichevsky and Gr{\'e}goire Bonnet},
  journal={Reports on Progress in Physics},
Fluorescence correlation spectroscopy (FCS) is an experimental technique using statistical analysis of the fluctuations of fluorescence in a system in order to decipher dynamic molecular events, such as diffusion or conformational fluctuations of biomolecules. First introduced by Magde et al to measure the diffusion and binding of ethidium bromide onto double-stranded DNA, the technique has been undergoing a renaissance since 1993 with the implementation of confocal microscopy FCS. Since then… 
Fluorescence Correlation Spectroscopy (FCS)
The specific features of FCS make it a versatile tool for biomolecular studies, including determination of translational and rotational diffusion, concentration and density of molecules, chemical kinetics, and binding reactions.
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This chapter presents a review of fluorescence correlation spectroscopy (FCS), an experimental technique with single-molecule sensitivity, which is based on the analysis of fluctuations of
Fluorescence correlation spectroscopy of autofluorescent proteins and its applications in live cell membranes
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Brief introduction to fluorescence correlation spectroscopy.
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  • 2013
Fluorescence correlation spectroscopy is a non-invasive technique and is based on fluctuations of fluorescence within a tiny observation volume under equilibrium conditions. With its wide
Fluorescence correlation spectroscopy in biology, chemistry, and medicine
The goal of this review is to promote the development of FCS in Russia so that this technique could occupy the position it deserves in modern Russian science.
Development and application of ultrasensitive fluorescence spectroscopy and microscopy for biomolecular interaction studies
This thesis describes the development of sensitive and high-resolution fluorescence spectroscopic and microscopic techniques and their application to probe biomolecules and their interactions in
Fluorescence correlation spectroscopy and allostery: the case of GroEL.
The utility of the FCS technique is exemplified in the case of the single-ring version of the Escherichia coli molecular chaperone GroEL that interconverts with relatively slow dynamics between two allosteric states: a T state with low affinity for ATP and an R state with high affinity forATP.
Two-dimensional fluorescence lifetime correlation spectroscopy. 1. Principle.
This work develops a new method that combines FCS and time-correlated single photon counting (TCPSC) to extract unambiguous information about equilibrium dynamics of complex molecular systems and performs a kinetic Monte Carlo simulation of a TCPSC-FCS experiment as a proof-of-principle example.


Fluorescence correlation microscopy of cells in the presence of autofluorescence.
Dynamics of fluorescence fluctuations in green fluorescent protein observed by fluorescence correlation spectroscopy.
FCS is a convenient, intrinsically calibrated method for pH measurements in subfemtoliter volumes with nanomolar concentrations of EGFP, and shows contributions from two chemical relaxation processes as well as diffusional concentration fluctuations.
Anomalous diffusion of fluorescent probes inside living cell nuclei investigated by spatially-resolved fluorescence correlation spectroscopy.
We have investigated spatial variations of the diffusion behavior of the green fluorescent protein mutant EGFP (F64L/S65T) and of the EGFP-beta-galactosidase fusion protein in living cells with
Fluorescence correlation spectroscopy. II. An experimental realization
This paper describes the first experimental application of fluorescence correlation spectroscopy, a new method for determining chemical kinetic constants and diffusion coefficients. These quantities
Fluorescence correlation spectroscopy of enzymatic DNA polymerization.
It is demonstrated that fluorescent 217-bp DNA fragments can be detected at very low initial ss M13mp18(+) DNA and tetramethylrhodamine-4-dUTP concentrations and that these polymers are cleaved by the chosen restriction enzymes.
Ultrasensitive hybridization analysis using fluorescence correlation spectroscopy.
Results of a simple homology search for the sequences complementary to the primer indicate the existence of additional sites of lower specificity, and suggested that approximately 2 primers can be associated with one template DNA at 40 degrees C.
Fluorescence-intensity distribution analysis and its application in biomolecular detection technology.
This work presents an extremely sensitive tool to monitor the interaction of fluorescently labeled molecules or other microparticles with their respective biological counterparts that should find a wide application in life sciences, medicine, and drug discovery.