Fluorescence spectroscopy takes advantage of quantum mechanical principles stating that some molecules are capable of absorbing electromagnetic radiation, this causes the molecule to enter an excited state. As the molecule transitions from the excited state back to the ground state it releases energy in the form of light, known as fluorescence. The wavelength of light required to excited a particle, and that is emitted by a particle is characteristic of that particle. Hence very few particles in a sample will usually fluoresce when excited at a given wavelength making fluorescence an extremely sensitive technique for the detection of particles. Under ideal conditions particle concentrations as low as one part per trillion can be detected, a significant increase in sensitivity over absorbance spectroscopy.
Fluorescence spectroscopy is capable of detecting not only fluorescent particles, but bio-luminescent and chemi-luminescent particles as well. Scanning of emission and excitation wave-lengths can be done to characterize the particle of interest under various conditions. Also an examination of the change in fluorescence intensity over time under a variety of conditions can provide valuable kinetic information and information about the stability of the particle of interest. In fact a variety of enzyme assays, cell proliferation assays, receptor binding studies, immunological assays, toxicity studies and quantitation assays take advantage of fluorescence detection and measurement.
At Analytical Ventura we have experience with a wide variety of fluorescence techniques and are happy to help you determine how to apply this powerful technique to your research.
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