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The term intrinsic DNA fluorescence refers to the fluorescence emitted directly by DNA when it absorbs ultraviolet (UV) radiation. It contrasts to that stemming from fluorescent labels that are either simply bound to DNA or covalently attached to it, [1] [2] widely used in biological applications; such labels may be chemically modified, not naturally occurring, nucleobases.
A simplified Jablonski diagram illustrating the change of energy levels.. The principle behind fluorescence is that the fluorescent moiety contains electrons which can absorb a photon and briefly enter an excited state before either dispersing the energy non-radiatively or emitting it as a photon, but with a lower energy, i.e., at a longer wavelength (wavelength and energy are inversely ...
Furthermore, tryptophan fluorescence is strongly influenced by the proximity of other residues (i.e., nearby protonated groups such as Asp or Glu can cause quenching of Trp fluorescence). Also, energy transfer between tryptophan and the other fluorescent amino acids is possible, which would affect the analysis, especially in cases where the ...
The zero-phonon line is located at a frequency ω’ determined by the intrinsic difference in energy levels between ground and excited state as well as by the local environment. The phonon sideband is shifted to a higher frequency in absorption and to a lower frequency in fluorescence.
The system relaxes vibrationally, and eventually fluoresces at a longer wavelength than the original high-energy photon had. The fluorescence lifetime refers to the average time the molecule stays in its excited state before emitting a photon. Fluorescence typically follows first-order kinetics:
Utilization of the intrinsic fluorescence properties of tryptophan residues in many proteins forms the basis of nanoDSF. The emission wavelengths of tryptophan residues are dependent on the surrounding chemical environment, notably solvation (see solvatochromism ) and therefore differ between folded and unfolded protein, just as with the ...
For example, if aluminum K-alpha X-rays are used, the intrinsic energy band has a FWHM of 0.43 eV, centered on 1,486.7 eV (E/ΔE = 3,457). If magnesium K-alpha X-rays are used, the intrinsic energy band has a FWHM of 0.36 eV, centered on 1,253.7 eV (E/ΔE = 3,483). These are the intrinsic X-ray line widths; the range of energies to which the ...
By detecting intrinsic fluorescence from aromatic residues such as tryptophan and tyrosine, researchers can assess environmental shifts that accompany structural transitions. Fluorescence data can be acquired alongside CD signals, particularly in temperature ramp experiments, where it helps monitor unfolding events by tracking changes in ...