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Fluorescence is one of two kinds of emission of light by a substance that has absorbed light or other electromagnetic radiation. When exposed to ultraviolet radiation, many substances will glow (fluoresce) with colored visible light. The color of the light emitted depends on the chemical composition of the substance.
Additionally, Fluorescence spectroscopy can be adapted to the microscopic level using microfluorimetry. In analytical chemistry, fluorescence detectors are used with HPLC. In the field of water research, fluorescence spectroscopy can be used to monitor water quality by detecting organic pollutants. [14]
A corollary of Kasha's rule is the Vavilov rule, which states that the quantum yield of luminescence is generally independent of the excitation wavelength. [4] [7] This can be understood as a consequence of the tendency – implied by Kasha's rule – for molecules in upper states to relax to the lowest excited state non-radiatively.
Jablonski diagram including vibrational levels for absorbance, non-radiative decay, and fluorescence. When a molecule absorbs a photon, the photon energy is converted and increases the molecule's internal energy level. Likewise, when an excited molecule releases energy, it can do so in the form of a photon.
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 ...
Jablonski diagram of FRET with typical timescales indicated. The black dashed line indicates a virtual photon.. Förster resonance energy transfer (FRET), fluorescence resonance energy transfer, resonance energy transfer (RET) or electronic energy transfer (EET) is a mechanism describing energy transfer between two light-sensitive molecules (chromophores). [1]
These techniques can be combined with microscopy, to map the intensity (confocal microscopy) or the lifetime (fluorescence-lifetime imaging microscopy) of the photoluminescence across a sample (e.g. a semiconducting wafer, or a biological sample that has been marked with fluorescent molecules). Modulated photoluminescence is a specific method ...
Fluorescence correlation spectroscopy (FCS) is a statistical analysis, via time correlation, of stationary fluctuations of the fluorescence intensity. Its theoretical underpinning originated from L. Onsager's regression hypothesis. The analysis provides kinetic parameters of the physical processes underlying the fluctuations.