Fluorescence is the emission of light from a molecule, which returns from the lowest vibration level of an excited single state to its normal ground state. Excited by the molecules, it is achieved by exposing the light source of the specified wavelength. At what time the molecule/compound is fluorescent, a part of the absorbed light is converted to fluorescence light, an emission of light at a lower wavelength. The fluorescence intensity is directly proportional to the concentration of the fluorescent species. Obviously, the linear relationship among the concentration and the fluorescent, intensity falls just to the diluted solution at high concentrations, make it required to first set up a concentration to detect the concentration of fluorescent graph and unknown sample with known standards. Fluorometry use to concentration determining of fluorophores, it provides sensitive absorption more than the colorimetric method.
Actually, there is a strong light source in a unit, a focusing system, sample compartment a primary filter to choose the suitable wavelength of the excitation source, secondary filter permits the transmission of fluorescence and absorbs the scattered excitation radiation, an electronic amplifier, a photocell, and the metering system. To reduce the interference of the primary source, fluorescence is collected at a 90-degree angle. A shutter permits the beam of the source to drop on the sample only throughout the measurement.
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