We can say that a Fluorometer is a special optical device commonly used in laboratory environments, which is capable of measuring the fluorescent quality of biological or mineral samples. Fluorescence occurs when a substance emits visible light and appears to glow after it has been exposed to some type of radiation, either alone or high-energy radiation such as x-rays of visible light.
The design of any typical fluorometer has several key components. It has an input source for ordinary visible light, and this light passes through an excitation filter that allows only the specific wavelengths of impacts at which a sample cell of the material was studied. When this material, whether organic or inorganic, is bombarded by these controlled wavelengths of light, it emits fluorescence, emitting characteristic light of its own that is then passed through an emission filter. The emissions are read by a light detector that produces a reading for the observer to know how the sample is reacting and what its content is.
Although fluorometer detection is based on the fundamental universal principles of fluorescence, there are several unique applications and adaptations for the devices. One of the main uses of fluorometers is that they are used to measure the chlorophyll fluorescence and thus investigate the physiology of plants, although they can currently have many applications.
What is fluorescence?
Fluorescence is a term that was first used in 1852 by George Gabriel Stokes. It is a particular type of luminescence that characterizes substances that are capable of absorbing energy in the form of electromagnetic radiation and then emitting part of that energy in the form of electromagnetic radiation of different wavelength.
The typical fluorescence mechanism involves three sequential steps: absorption, non-radiative dissipation and emission.
What factors affect fluorescence?
At low concentrations, the fluorescence intensity is generally proportional to the concentration of the fluorophore.
Unlike ‘standard’ visible or ultraviolet spectrometry, device-independent spectra are not easy to reach. There are several factors that influence and distort the spectra, and corrections are necessary to achieve the ‘true’ spectrum, that is, independent of the machine.
The fluorescence intensity is affected by the following factors: structure, temperature and nature of the solvent, effect of pH, effect of dissolved oxygen.
What is fluorescence spectrometry based on?
Fluorescence spectrometry (also called fluorometry or spectrofluorimetry) is a type of electromagnetic spectroscopy that analyzes the fluorescence of a sample. It involves using a beam of light, usually ultraviolet light, that excites the electrons of the molecules of certain compounds and causes them to emit light of lower energy, usually visible light (although not necessarily). A complementary technique is absorption spectrometry. Devices that measure fluorescence are calledfluorometers or fluorimeters. Fluorescence spectrometry is used in biochemical, medical, chemical and research analyzes of organic compounds.
What is the most recent application of fluorometers?
Fluorescence spectrometry is used in biochemical, medical, chemical and research analyzes of organic compounds. It has also been used to differentiate malignant tumors from benign skin.
Currently one of the most modern applications of fluorometry is the quantification of nucleic acids, through fluorometers, which measure concentrations of DNA, RNA, and proteins with high precision and sensitivity. It is based on the use of fluorophores that are specifically interspersed between the molecules of interest, thus minimizing the effects of contaminants. The accuracy of the measurements even at low concentrations (range of effectiveness from 5 ng to 1 ug) which makes these devices an ideal tool for applications such as real-time PCR and mass sequencing.
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