Fluorescence resonance energy transfer FRET efficiency

Fluorescence resonance energy transfer (FRET) has also been used very often to design optical sensors. In this case, the sensitive layer contains the fluorophore and an analyte-sensitive dye, the absorption band of which overlaps significantly with the emission of the former. Reversible interaction of the absorber with the analyte species (e.g. the sample acidity, chloride, cations, anions,...) leads to a variation of the absorption band so that the efficiency of energy transfer from the fluorophore changes36 In this way, both emission intensity- and lifetime-based sensors may be fabricated. 

The elucidation of the structure, dynamics and self assembly of biopolymers has been the subject of many experimental, theoretical and computational studies over the last several decades. [1, 2] More recently, powerful singlemolecule (SM) techniques have emerged which make it possible to explore those questions with an unprecedented level of detail. [3-55] SM fluorescence resonance energy transfer (FRET), [56-60] in particular, has been established as a unique probe of conformational structure and dynamics. [26-55] In those SM-FRET experiments, one measures the efficiency of energy transfer between a donor dye molecule and an acceptor dye molecule, which label specific sites of a macromolecule. The rate constant for FRET from donor to acceptor is assumed to be given by the Forster theory, namely [59,61-64]... 

Forster s theory [1], has enabled the efficiency of EET to be predicted and analyzed. The significance of Forster s formulation is evinced by the numerous and diverse areas of study that have been impacted by his paper. This predictive theory was turned on its head by Stryer and Haugland [17], who showed that distances in the range of 2-50 nm between molecular tags in a protein could be measured by a spectroscopic ruler known as fluorescence resonance energy transfer (FRET). Similar kinds of experiments have been employed to analyze the structure and dynamics of interfaces in blends of polymers. 

The fluorescence resonance energy transfer (FRET) quenching efficiency is defined as ... 

Fluorescence resonance energy transfer (FRET) experiments commonly use the fluorescent spectrum and relaxation times of the Forster donor and acceptor chromophores to find the distances between fluorescent dyes at labeled sites in protein, DNA, RNA, etc. FRET is a type of spectroscopic ruler . The computation uses either experimental quantum yields or relaxation lifetimes to calculate the efficiency of resonance energy transfer Ej. 

Fluorescence Resonance Energy Transfer (FRET), Fig. 1 (a) Jablonski diagram illustrating FRET and related processes, including excitation of the donor, radiative (solid line) and non-radiative (dashed lines) relaxation on the donor and acceptor, vibrational relaxation (short curved arrows), and transitions associated with FRET (dotted lines). Processes that determine the FRET efficiency are indicated in bold, (b) Illustration of spectral overlap between Cy3 (donor) emission and Cy5 (acceptor) absorption, (c) Definition of the angles used to calculate... 

Fluorescence Resonance Energy Transfer (FRET), Fig. 1 (continued) the orientation factor from the relative alignment of the donor and acceptor transition dipoles, (d) ERET efficiency as a function of the relative distance between the donor and acceptor... 

Fluorescence Resonance Energy Transfer (FRET), Fig. 2 Model data for the change in the emission spectra for a Cy3 donor paired with a Cy5 acceptor (1 1 ratio) at different FRET efficiencies (0 %, 33%, 61 %). Note the small direct excitation of Cy5 in the absence of FRET. Increases in FRET efficiency are reflected by progressive quenching of the Cy3 fluorescence and sensitization of Cy5 fluorescence. The model data is for excitation at 520 nm, where 520, cy3 = 79,500 cm- ... 

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