Dynamic quenching as a simple test for the mechanism of excited-state reaction

Томин, В. И.; Oncul, Sule; Smolarczyk, Grzegorz; Demchenko, Alexander P.

doi:10.1016/j.chemphys.2007.09.036

Cited by 48 publications

(30 citation statements)

References 36 publications

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“…Since the dynamic equilibrium between these two forms is established prior to emission (Shynkar et al 2003), the factors changing the fluorescence lifetime (temperature, dynamic quenchers) do not change the ratiometric response Tomin et al 2007). Both these bands are strongly Stokes-shifted, so the ''blue'' excitation (with the maximum at about 420 nm) allows avoiding lightscattering artefacts.…”

Section: The Methods Based On F2n12s Probementioning

confidence: 99%

Beyond annexin V: fluorescence response of cellular membranes to apoptosis

2012

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Dramatic changes in the structure of cell membranes on apoptosis allow easy, sensitive and non-destructive analysis of this process with the application of fluorescence methods. The strong plasma membrane asymmetry is present in living cells, and its loss on apoptosis is commonly detected with the probes interacting strongly and specifically with phosphatidylserine (PS). This phospholipid becomes exposed to the cell surface, and the application of annexin V labeled with fluorescent dye is presently the most popular tool for its detection. Several methods have been suggested recently that offer important advantages over annexin V assay with the ability to study apoptosis by spectroscopy of cell suspensions, flow cytometry and confocal or twophoton microscopy. The PS exposure marks the integrated changes in the outer leaflet of cell membrane that involve electrostatic potential and hydration, and the attempts are being made to provide direct probing of these changes. This review describes the basic mechanisms underlying the loss of membrane asymmetry during apoptosis and discusses, in comparison with the annexin V-binding assay, the novel fluorescence techniques of detecting apoptosis on cellular membrane level. In more detail we describe the detection method based on smart fluorescent dye F2N12S incorporated into outer leaflet of cell membrane and reporting on apoptotic cell transformation by easily detectable change of the spectral distribution of fluorescent emission. It can be adapted to any assay format.

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Section: The Methods Based On F2n12s Probementioning

confidence: 99%

Beyond annexin V: fluorescence response of cellular membranes to apoptosis

2012

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“…In the case of rapidly established equilibrium between two forms, the internally calibrated signal is resistant to any uncontrolled quenching effect produced by collisional quenchers or the temperature. This is because their lifetimes are equal and they are quenched proportionally with the retention of the same intensity ratio [54][55][56]. Since the ESIPT reaction can provide dramatic shifts in fluorescence spectra (by 100 nm and more), finding new systems exhibiting dual emission based on ESIPT is a great concern.…”

Section: Transitions Between Excited-state Formsmentioning

confidence: 99%

Comparative Analysis of Fluorescence Reporter Signals Based on Intensity, Anisotropy, Time-Resolution, and Wavelength-Ratiometry

Demchenko

2010

Springer Series on Fluorescence

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The response signal of an immense number of fluorescence reporters with a broad variety of structures and properties can be realized through the observation in changes of a very limited number of fluorescence parameters. They are the variations in intensity, anisotropy (or polarization), lifetime, and the spectral changes that allow wavelength-ratiometric detection. Here, these detection methods are overviewed, and specific demands addressed to fluorescence emitters for optimization of their response are discussed.

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“…A kinetic barrier may appear owing to the change in the relative configuration (see Figure 22.1) of the phenyl and chromone rings (which may be coupled with the solvation coordinate [28,59]), Figure 22.6 Scheme of two mechanisms of the ESIPT reaction: upper scheme -kinetic (slow irreversible) reaction; lower scheme -fast reversible reaction. Reprinted with permission from [87]. Copyright Elsevier and, in protic environments, owing to competition between intramolecular and intermolecular hydrogen bonds [58].…”

Section: Excited-state Transformations and Their Modellingmentioning

confidence: 99%

“…In Ref. [87] an original method based on the dynamic quenching of fluorescence by the nitric oxide spin compound TEMPO was proposed. For different dyes, TEMPO exhibits very high bimolecular quenching constants that are close to the diffusional limit.…”

Section: Excited-state Transformations and Their Modellingmentioning

confidence: 99%

“…For different dyes, TEMPO exhibits very high bimolecular quenching constants that are close to the diffusional limit. Tomin et al [87] developed the theory, describing the effects of dynamic quenching within the framework of two-state excited-state reaction formalism, which predicts interesting, non-trivial behaviour of two-band emission in steady-state spectra, and carried out experiments on the quenching by TEMPO of the fluorescence of several 3-hydroxychromone dyes that exhibit excited-state intramolecular proton transfer reaction and on which the two types of reaction mechanism are observed. These results are in line with the predictions of theory.…”

Section: Excited-state Transformations and Their Modellingmentioning

confidence: 99%

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