Photochemistry
Fluorescence occurs when a molecule, atom or nanostructure relaxes to its ground state after being electrically excited
Excitation: so+hvex→ s1
Fluorescence (emission): s1→ so + hvem here hν is a generic term for photon energy with h = Planck's constant and ν = frequency of light. (The specific frequencies of exciting and emitted light are dependent on the particular system.)
State S0 is called the ground state of the fluorophore (fluorescent molecule) and S1 is its first electronically) excited state
Relaxation of an S1 state can also occur through interaction with a second molecule through fluorescence quenching. Molecular oxygen (O2) is an extremely efficient quencher of fluorescence because of its unusual triplet ground state
Molecules that are excited through light absorption or via a different process (e.g. as the product of a reaction) can transfer energy to a second 'sensitized' molecule, which is converted to its excited state and can then fluoresce. This process is used in lightsticks to produce different colors
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
Applications
Lighting
In the mid 1990s, white light-emitting diodes (LEDs) became available, which work through a similar process. Typically, the actual light-emitting semiconductor produces light in the blue part of the spectrum, which strikes a phosphor compound deposited on the chip; the phosphor fluoresces from the green to red part of the spectrum. The combination of the blue light that goes through the phosphor and the light emitted by the phosphor produce a net emission of white light
Glow sticks sometimes utilize fluorescent materials to absorb light from the chemiluminescent reaction and emit light of a different color
Fluorescence in several wavelengths can be detected by an array detector, to detect compounds from HPLC flow. Also, TLC plates can be visualized if the compounds or a coloring reagent is fluorescent. Fluorescence is most effective when there is a larger ratio of atoms at lower energy levels in a Boltzmann distribution. There is then a higher probability of lower energy atoms being excited and releasing photons, making analysis more efficient
Fingerprints can be visualized with fluorescent compounds such as ninhydrin
Fluorescence in the life sciences is used generally as a non-destructive way of tracking or analysis biological molecules by means of the fluorescent emission at a specific frequency were there is no background from the excitation light, as relatively few cellular components are naturally fluorescent (called intrinsic or autofluorescence). In fact, a protein or other component can be "labelled" with a extrinsic fluorophore, a fluorescent dye which can be a small molecule, protein or quantum dot, finding a large use in many biological applications. The quantification of a dye is done with a Spectrofluorometer and finds additional applications in:the following
when scanning the fluorescent intensity across a plane one has Fluorescence microscopy of tissues, cells or subcellular structures which is accomplished by labeling an antibody with a fluorophore and allowing the antibody to find its target antigen within the sample. Labeling multiple antibodies with different fluorophores allows visualization of multiple targets within a single image (multiple channels). DNA microarrays are a variant of this
Automated sequencing of DNA by the chain termination method; each of four different chain terminating bases has its own specific fluorescent tag. As the labeled DNA molecules are separated, the fluorescent label is excited by a UV source, and the identity of the base terminating the molecule is identified by the wavelength of the emitted light
. Ethidium bromide fluoresces orange when intercalating DNA and when exposed to UV light
FACS) fluorescent-activated cell sorting)
DNA detection: the compound ethidium bromide, when free to change its conformation in solution, has very little fluorescence. Ethidium bromide's fluorescence is greatly enhanced when it binds to DNA, so this compound is very useful in visualising the location of DNA fragments in agarose gel electrophoresis. Ethidium bromide can be toxic - a safer alternative is the dye SYBR Green
Immunology: An antibody has a fluorescent chemical group attached, and the sites (e.g., on a microscopic specimen) where the antibody has bound can be seen, and even quantified, by the fluorescence
Additionally Fluorescence resonance energy transfer used to study protein interactions, detect specific nucleic acid sequences and used as biosensors, while fluorescent lifetime can give an additional layer of information
Gemology, mineralogy, geology, and forensics
Gemstones, minerals, fibers, and many other materials which may be encountered in forensics or with a relationship to various collectibles may have a distinctive fluorescence or may fluoresce differently under short-wave ultraviolet, long-wave ultra violet, or X-rays
Many types of calcite and amber will fluoresce under shortwave UV. Rubies, emeralds, and the Hope Diamond exhibit red fluorescence under short-wave UV light; diamonds also emit light under X ray radiation
Crude oil (petroleum) fluoresces in a range of colors, from dull brown for heavy oils and tars through to bright yellowish and bluish white for very light oils and condensates. This phenomenon is used in oil exploration drilling to identify very small amounts of oil in drill cuttings and core samples
Organic liquids
Organic liquids such as mixtures of anthracene in benzene, toluene, or stilbene in the same solvents, fluoresce with ultraviolet or gamma ray irradiation. The decay times of this fluorescence is of the order of nanoseconds since the duration of the light depends on the lifetime of the excited states of the fluorescent material, in this case anthracene or stilbene
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