Time-Domain Lifetime Measurements

  • Joseph R. Lakowicz


Time-resolved measurements are widely used in fluorescence spectroscopy, particularly for studies of biological macromolecules. This is because time-resolved data frequently contain more information than is available from the steady-state data. For instance, consider a protein which contains two tryptophan residues, each with a distinct lifetime. Because of spectral overlap of the absorption and emission, it is not usually possible to resolve the emission from the two residues. However, the time-resolved data may reveal two decay times, which can be used to resolve the emission spectra and relative intensities of the two tryptophan residues. Then one can question how each of the tryptophan residues is affected by the interactions of the protein with its substrate or other macromolecules. Is one of the tryptophan residues close to the binding site? Is a tryptophan residue in a distal domain affected by substrate binding to another domain? Such questions can be answered if one measures the decay times associated with each tryptophan residue.


Decay Time Fluorescence Decay Lifetime Measurement Anthranilic Acid Streak Camera 
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Copyright information

© Springer Science+Business Media New York 1999

Authors and Affiliations

  • Joseph R. Lakowicz
    • 1
  1. 1.University of Maryland School of MedicineBaltimoreUSA

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