Advantages and disadvantages of fluorescence spectroscopy

The fluorescence spectroscopy is a type of spectroscopy and is also called fluorometry. It is used to identify and measure the concentrations of analytes in a sample. The processes involve the excitation of electrons in molecules of a particular analyte by a beam of light (ultraviolet) and encourage them to emit light (visible). It is a molecular absorption of light energy at a wavelength and its almost immediate re-emission at a different, longer wavelength. Some components are fluorescent naturally, and others should be modified for fluorescence.
The advantages of fluorescence spectroscopy are as follows.

• Its high sensitivity is the main advantage of fluorometry.
• Due to the unique optical properties of the component, it has high specificity.
• It can determine fluorescence intensity, decay time, and the concentration of the component.
• It may immune to the scattering of light.
• The emitted light is read at the right angle to the exciting light, reducing the background signal
• These types of methods have a large range of linearity.

The disadvantages of fluorescence spectroscopy are as follows.

• The major disadvantage of fluorescence spectroscopy is that not all molecules are fluorescent.
• It has limitations related to loss of recognition capability and photostability.
• Susceptible to interference because of the changes in pH and oxygen levels of the sample.
• It is susceptible to the auto-fluorescence of the sample.
• Issue related to potential toxicity, due to the foreign material in the biological media.
• The short lifespan of fluorophores is another disadvantage of fluorometry.

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