Why fluorescence spectroscopy

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Skip to content Fluorescence is the phenomenon where a molecule absorbs light within its absorption band and then emits this light at longer wavelengths within its emission band.

Fluorescence Spectroscopy Configuration In fluorescence spectroscopy, a light of a specific wavelength band is passed through a solution, which emits the light towards a filter and into a detector for measurement.

Fluorescence Spectroscopy Filters We briefly mentioned filters when outlining the configuration of a fluorescence spectroscopy device, and most of these devices are based on the filters within them. We use cookies to ensure that we give you the best experience on our website. If you continue to use this site we will assume that you are ok with it. Accept Read More. Privacy Policy. Close Privacy Overview This website uses cookies to improve your experience while you navigate through the website.

Once many fluorescent substances stop the incident light, the luminescence phenomenon will disappear immediately. The emitted light with this property is called fluorescence. When light irradiates certain atoms, the energy of the light causes some electrons around the nucleus to transition from the original orbit to a higher energy orbit, that is, transition from the ground state to the first excited singlet state or the second excited singlet state.

Electrons at both states are unstable, so they will return to the ground state. And when they do, the energy is released in the form of light, generating fluorescence. Any fluorescent compound has two characteristic spectra: excitation spectrum and emission spectrum. The excitation spectrum reflects the dependence of the measured fluorescence intensity on the excitation wavelength at a fixed emission wavelength. We call it phosphorescence if the emission takes longer: in the order of milli- seconds.

This is shown in the following 4 minute video. Only a relatively small number of compounds can fluoresce. Some non-fluorescent compounds can be made fluorescent by adding a fluorescent label. In general molecules that fluoresce have one or more aromatic groups in its structure. A molecule can be excited from its electronic ground state. In the electronic ground state the molecule has the lowest possible electronic energy.

Upon excitation the absorption of a photon one of the electrons goes into an higher electronic state and the molecule is excited. The molecule will stay in its electronic excited state in the order of pico or nanoseconds ns. Then the electron will fall back to its ground state and will emit a photon of a longer wavelength than the photon used for excitation. Emission after excitation with UV light Different fluorophores, their structure and the emission. Your browser does not support JavaScript!

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