The technique of fluorescence spectroscopy is considered to be an analytical one that pays attention to the fluorescence properties of the sample and allows carrying out quantitative measurements of drugs, metabolites, and other chemical products. Thus, the fluorescence used by fluorescence process spectroscopy helps to analyze samples, to be more precise, the fluorophore assimilates light of a specific wavelength that then excites the electron, which changes from ground state to a high energy excited state.
Fluorescence spectroscopy offers a highly important advantage as quantum efficiency because it provides a measurement process of photon vs converted electrons, or the number of incident photons is absorbed vs the number converted to electrons. Also, it should be mentioned that this parameter plays a great role in the technology of process spectroscopy because it has a high level in samples at low concentration.
Nonetheless, there are several factors that can influence on fluorescence spectroscopy:
- Molecular rigidity. The thing is that the technique of fluorescence spectroscopy requires fluorophores with rigid structures because it helps to reduce vibrations and the possibility of converting in the triplet state.
- Solvent polarity. The fluorescence degree is dependent on the solvent polarity since the presence of heavy atoms reduces the fluorescence during process spectroscopy.
- Dissolved oxygen reduces the emission intensity during fluorescence spectroscopy.
- pH level, in its turn, affects the fluorescent qualities.
- Quenching process is able to decrease in the fluorescence intensity.
The principle of fluorescence spectroscopy operation is based on the use of a light source (xenon or mercury arc lamps), monochromator and a detector. The process spectroscopy application includes the following fields: fluorescence probes where the technology of process spectroscopy allows detecting qualitatively and quantitatively biological compounds present in low concentration; protein structure can be determined by fluorescence process spectroscopy; fluorescence-activated cell sorting that allows sorting various cells based on the differences in fluorescent properties.
Finally, it should be noted that there is a probability of high susceptibility of fluorescence spectroscopy technique to the conditions mentioned above. The changes can be made by even interfering materials, thus, fluorescence can be lost during the interaction with other molecules, basically, in concentrated samples.
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