ABSTRACT
Fluorescence spectroscopy has been used for a long time as an efficient tool in the fields of molecular biology, biophysics, and biochemistry. This chapter describes basic principles of fluorescence, types of fluorescence spectra, intrinsic and extrinsic fluorophores, the factors affecting fluorescence, including concentration, solvent and local environment, quenching, light scattering, and temperature, as well as several approaches to obtain fluorescence data. The general principles of light absorption and emission are illustrated by a Jablonski diagram. The wide range of closely spaced excitation wavelengths associated with excitation transitions in fluorescent molecule leads to a broad excitation band rather than discrete lines. Emission-excitation matrix fluorescence spectroscopy is known as a powerful method for the analysis of complex mixtures because total luminescence spectrum represent the total fluorescence profiles of the samples. Synchronous fluorescence spectrum is obtained by scanning both the excitation and emission monochromators simultaneously, usually in such a way that a constant wavelength interval is kept between them.
