FLUORESCENCE Fluorescence is the emission of light by a substance that has absorbed light or other electromagnetic radiation. It is a form of luminescence.

In most cases, th e emitted light has a longer wavelength, and therefore lower energy, than the ab sorbed radiation. However, when the absorbed electromagnetic radiation is intens e, it is possible for one electron to absorb two photons; this two-photon absorp tion can lead to emission of radiation having a shorter wavelength than the abso rbed radiation. The emitted radiation may also be of the same wavelength as the absorbed radiation, termed "resonance fluorescence".[1] The most striking examples of fluorescence occur when the absorbed radiation is in the ultraviolet region of the spectrum, and thus invisible to the human eye, and the emitted light is in the visible region. Fluorescence has many practical applications, including mineralogy, gemology, ch emical sensors (fluorescence spectroscopy), fluorescent labelling, dyes, biologi cal detectors, and, most commonly, fluorescent lamps. fluorphore A fluorophore (or fluorochrome, similarly to a chromophore) is a fluorescent che mical compound that can re-emit light upon light excitation. Fluorophores typica lly contain several combined aromatic groups, or plane or cyclic molecules with several p bonds. Fluorophores are sometimes used alone, as a tracer in fluids, as a dye for stain ing of certain structures, as a substrate of enzymes, or as a probe or indicator (when its fluorescence is affected by environment such as polarity, ions,...). But more generally it is covalently bonded to a macromolecule, serving as a mark er (or dye, or tag, or reporter) for affine or bioactive reagents (antibodies, p eptides, nucleic acids). Fluorophores are notably used to stain tissues, cells, or materials in a variety of analytical methods, i.e. fluorescent imaging and sp ectroscopy. Fluorescein, by its amine reactive isothiocyanate derivative FITC, has been one of the most popularized fluorophores. From antibody labeling, the applications h ave spread to nucleic acids thanks to (FAM(Carboxyfluorescein), TET,...). Other historically common fluorophores are derivatives of rhodamine (TRITC), coumarin, and cyanine.[1] Newer generations of fluorophores, many of which are proprietar y, often perform better (more photostable, brighter, and/or less pH-sensitive) t han traditional dyes with comparable excitation and emission.[2][3] QUENCHING

Quenching refers to any process which decreases the fluorescence intensity of a given substance. A variety of processes can result in quenching, such as excited state reactions, energy transfer, complex-formation and collisional quenching. As a consequence, quenching is often heavily dependent on pressure and temperatu re. Molecular oxygen, iodide ions and acrylamide[1] are common chemical quencher s. The chloride ion is a well known quencher for quinine fluorescence. [2] [3] [ 4] Quenching poses a problem for non-instant spectroscopic methods, such as lase r-induced fluorescence. Quenching is made use of in optode sensors; for instance the quenching effect of oxygen on certain ruthenium complexes allows the measurement of oxygen saturati on in solution. Quenching is the basis for fluorescence resonance energy transfe r (FRET) assays.[5][6][7] Quenching and dequenching upon interaction with a spec ific molecular biological target is the basis for activatable optical contrast a gents for molecular imaging.[8][9]