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Time-Resolved Protein Fluorescence

  • Chapter
Principles of Fluorescence Spectroscopy

Abstract

In the previous chapter we presented an overview of protein fluorescence. We described the spectral properties of the aromatic amino acids and how these properties depend on protein structure. We now extend this discussion to include time-resolved measurements of intrinsic protein fluorescence. Prior to 1983, most measurements of time-resolved fluorescence were performed using TCSPC. The instruments employed for these measurements typically used a flashlamp excitation source and a standard dynodechain-type PMT. Such instruments provided instrument response functions with a half-width near 2 ns, which is comparable to the decay time of most proteins. The limited repetition rate of the flashlamps, near 20 kHz, resulted in data of modest statistical accuracy, unless the acquisition times were excessively long. Given the complexity of protein intensity and anisotropy decays, and the inherent difficulty of resolving multiexponential processes, it was difficult to obtain definitive information on the decay kinetics of proteins.

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    Joseph R. Lakowicz

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Lakowicz, J.R. (1999). Time-Resolved Protein Fluorescence. In: Principles of Fluorescence Spectroscopy. Springer, Boston, MA. https://doi.org/10.1007/978-1-4757-3061-6_17

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  • DOI: https://doi.org/10.1007/978-1-4757-3061-6_17

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